Cancer represents a significant disease that profoundly impacts human health and longevity.Projections indicate a 47%increase in the global cancer burden by 2040 compared to 2020,accompanied by a further rise in the a...Cancer represents a significant disease that profoundly impacts human health and longevity.Projections indicate a 47%increase in the global cancer burden by 2040 compared to 2020,accompanied by a further rise in the associated economic burden.Consequently,there is an urgent need to discover and develop new alternative drugs to mitigate the global impact of cancer.Natural products(NPs)play a crucial role in the identification and development of anticancer therapeutics.This study identified ustusolate E(UE)and its analog 11α-hydroxy-ustusolate E(HUE)from strain Aspergillus calidoustus TJ403-EL05,and examined their antitumor activities and mechanisms of action.The findings demonstrate that both compounds significantly inhibited the proliferation and colony formation of AGS(human gastric cancer cells)and 786-O(human renal clear cell carcinoma cells),induced irreversible DNA damage,blocked the cell cycle at the G_(2)/M phase,and further induced apoptosis in tumor cells.To the best of the authors’knowledge,this is the first report on the anticancer effects of UE and HUE and their underlying mechanisms.The present study suggests that HUE and UE could serve as lead compounds for the development of novel anticancer drugs.展开更多
Objective:The occurrence and development of atrial fibrillation(AF)are influenced by the autonomic nervous system and inflammation.Acupuncture is an effective treatment for AF.This study explored the protective effect...Objective:The occurrence and development of atrial fibrillation(AF)are influenced by the autonomic nervous system and inflammation.Acupuncture is an effective treatment for AF.This study explored the protective effects of acupuncture in a rat model of paroxysmal AF and investigated its mechanisms.Methods:Male Sprague-Dawley rats(n=130)were randomly divided into blank control(Con),sham operation(Sham),AF,and acupuncture treatment(Acu)groups.A paroxysmal AF model was established by rapid atrial pacing through the jugular vein.Rats in the Acu group were immobilized to receive acupuncture treatment at Neiguan acupoint(PC6)for 20 min daily for seven days.The other groups were immobilized for the same duration over the treatment period but did not receive acupuncture.The AF induction rate,AF duration,cardiac electrophysiological parameters,and heart rate variability were evaluated by monitoring surface electrocardiogram and vagus nerve discharge signals.After the intervention,the rats were euthanized,and atrial morphology was assessed using haematoxylin and eosin staining.The expression of macrophage F4/80 antigen(F4/80)and cluster of differentiation(CD)86 in atrial myocardial tissue was detected using immunohistochemistry,immunofluorescence and flow cytometry.The expression levels or contents of interleukin(IL)-1β,IL-6,tumor necrosis factor-a(TNF-a),a7 nicotinic acetylcholine receptor(a7nAChR),phosphorylated Janus kinase 2(p-JAK2),and phosphorylated signal transducer and activator of transcription 3(p-STAT3)in atrial myocardial tissue were detected using Western blotting,reverse transcription-quantitative polymerase chain reaction,or enzyme-linked immunosorbent assay.The role of a7nAChR in acupuncture treatment was verified by intraperitoneal injection of the a7nAChR antagonist methyllycaconitine(MLA).Results:Compared with the AF group,acupuncture significantly reduced AF duration and induction rate,improved cardiac electrophysiology by enhancing vagus nerve activity and regulating autonomic balance.It also decreased the pro-inflammatory M1 macrophage proportion,alleviating myocardial injury and infiltration.MLA weakened acupuncture's electrophysiological improvement and anti-inflammatory effect.Results suggest that acupuncture triggers the a7nAChR-JAK2/STAT3 pathway and exerts cardioprotection via neuroimmune regulation.Conclusion:Acupuncture significantly reduced the AF induction rate,shortened AF duration,improved cardiac electrophysiological parameters,enhanced vagus nerve activity,and decreased the expression of pro-inflammatory M1 macrophages and inflammatory factors in rats with paroxysmal AF.展开更多
Background:Gallic acid(GA),a plant-derived polyphenol,possesses diverse biological functions such as reducing inflammation and against tumors.Currently,the influence of GA on the resistance of esophageal squamous cell...Background:Gallic acid(GA),a plant-derived polyphenol,possesses diverse biological functions such as reducing inflammation and against tumors.Currently,the influence of GA on the resistance of esophageal squamous cell carcinoma(ESCC)cells to cisplatin(DDP)is not well understood.Methods:Cell counting kit-8 assay examined how GA affected KYSE30 and TE-1 cell viability.5-Ethynyl-2′-deoxyuridine and TdT-mediated dUTP Nick-End labeling staining detected cell proliferation and apoptosis.Clone formation assay,flow cytometry,Carboxyfluorescein diacetate succinimidyl ester fluorescent probes,and Transwell assay determined cell biological properties,and 2′,7′-Dichlorofluorescin diacetate(DCFH-DA)fluorescent probes detected oxidative stress levels.Signal transducer and activator of transcription 3(STAT3)/Notch pathway protein levels after GA and/or Interleukin-6(IL-6)intervention were examined through Western blot.Furthermore,a model for subcutaneous graft tumors was established in nude mice.Results:GA exerted suppressive effects on cell proliferation,and caused apoptosis of KYSE30 and TE-1 cells.IL-6 intervention activated the STAT3/Notch pathway and promoted the malignant biological properties of ESCC cells.In contrast,GA attenuated the effects of IL-6,while STAT3 or Notch inhibitor further enhanced the effects of GA,suggesting that GA inhibited the IL-6/STAT3/Notch pathway.Not only that,GA promoted oxidative stress and enhanced cell sensitivity to DDP both in vitro and in vivo.Conclusion:GA suppresses the malignant progression of ESCC and enhances cell sensitivity to DDP by hindering the IL-6/STAT3/Notch pathway.展开更多
BACKGROUND Excessive endoplasmic reticulum(ER)stress in intestinal epithelial cells can lead to damage to the intestinal mucosal barrier,activate the signal transducer and activator of transcription 3(STAT3)/nuclear f...BACKGROUND Excessive endoplasmic reticulum(ER)stress in intestinal epithelial cells can lead to damage to the intestinal mucosal barrier,activate the signal transducer and activator of transcription 3(STAT3)/nuclear factor kappa B(NF-κB)signaling pathway,and exacerbate the inflammatory response,thus participating in the pathogenesis of ulcerative colitis(UC).Mesalazine is a commonly used drug in the clinical treatment of UC.However,further studies are needed to determine whether mesalazine regulates the ER stress of intestinal epithelial cells,downregulates the STAT3/NF-κB pathway to play a role in the treatment of UC.AIM To study the therapeutic effects of mesalazine on spontaneous colitis in interleukin-10(IL-10)-/-mice.METHODS The 24-week-old IL-10-/-mice with spontaneous colitis were divided into the model group and the 5-amino salicylic acid group.Littermates of wild-type mice of the same age group served as the control.There were eight mice in each group,four males and four females.The severity of symptoms of spontaneous colitis in IL-10-/-mice was assessed using disease activity index scores.On day 15,the mice were sacrificed.The colon length was measured,and the histopathological changes and ultrastructure of colonic epithelial cells were detected.The protein expressions of STAT3,p-STAT3,NF-κB,IκB,p-IκB,and glucoseregulated protein 78 were identified using Western blotting.The STAT3 and NF-κB mRNA expressions were identified using real-time polymerase chain reaction.The glucose-regulated protein 78 and C/EBP homologous protein expressions in colon sections were detected using immunofluorescence.RESULTS Mesalazine reduced the symptoms of spontaneous colitis in IL-10 knockout mice and the histopathological damage of colonic tissues,and alleviated the ER stress in epithelial cells of colitis mice.Western blotting and quantitative real-time polymerase chain reaction results showed that the STAT3/NF-κB pathway in the colon tissue of model mice was activated,suggesting that this pathway was involved in the pathogenesis of UC and might become a potential therapeutic target.Mesalazine could down-regulate the protein expressions of p-STAT3,NF-κB and p-IκB,and down-regulate the mRNA expression of STAT3 and NF-κB.CONCLUSION Mesalazine may play a protective role in UC by reducing ER stress by regulating the STAT3/NF-κB signaling pathway.展开更多
AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)...AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.展开更多
Objective:To investigate the biological functions and molecular regulatory mechanisms of kinesin family member 11(KIF11)in colorectal cancer(CRC).Methods:The expression of KIF11 in CRC was examined by qRT⁃PCR and publ...Objective:To investigate the biological functions and molecular regulatory mechanisms of kinesin family member 11(KIF11)in colorectal cancer(CRC).Methods:The expression of KIF11 in CRC was examined by qRT⁃PCR and public databases.Functional assays(CCK⁃8,colony formation,EdU,and Transwell)were employed to evaluate KIF11’s roles in CRC progression.Western blot,RIP⁃qPCR,MeRIP⁃qPCR,and RNA stability assays were performed to elucidate the molecular mechanism of N6⁃methyladenosine(m6A)modification for KIF11.RNA sequencing(RNA⁃seq)and correlation analysis were used to examine the downstream mechanism of KIF11 regulation.Results:KIF11 was highly expressed in CRC and promoted CRC proliferation and migration.Mechanistically,methyltransferase⁃like 3(METTL3)/insulin like growth factor 2 mRNA binding protein 2(IGF2BP2)enhanced KIF11 mRNA stability and expression in an m6A⁃dependent way.Furthermore,by means of the PROM1/PI3K/AKT pathway,KIF11 facilitated the progression of CRC.Conclusion:The m6A modification of KIF11 by METTL3/IGF2BP2 contributes to CRC progression via the PI3K/AKT signaling pathway,highlighting its potential as a prognostic biomarker and therapeutic target.展开更多
OBJECTIVE To explore hypoglycemic effect of 95%ethanol fraction of Nitraria roborowskii Kom(NRK-C)and its possible mechanism evaluated in the type 2 diabetes mellitus(T2DM)mice.METHODS The body weight,organ indices,bl...OBJECTIVE To explore hypoglycemic effect of 95%ethanol fraction of Nitraria roborowskii Kom(NRK-C)and its possible mechanism evaluated in the type 2 diabetes mellitus(T2DM)mice.METHODS The body weight,organ indices,blood glucose levels,serum biochemical indexes,as well as HE/PAS histopathological section were all analyzed to assess the hypoglycemic effect of NRK-C in T2DM mice induced by a high-fat diet(HFD)combined with six intraperitoneal injections of 35 mg·kg^(-1)of streptozotocin(STZ).The Western blotting and immunofluorescence were further applied to determine the regulatory effect of NRK-C on key signaling proteins.RESULTS The fasting blood glucose levels were significantly reduced after 7 weeks of administration of NRK-C.In addition,NRK-C could also significantly improve glucose tolerance,hepatic glycogen levels,and lipid levels(total cholesterol,triglyceride,low density lipoprotein and high density lipoprotein),and significantly reduced insulin resistance of diabetic mice,which played an important role in the antidiabetic effects.Further mechanism research demonstrated that phosphorylated PI3K expression was up-regulated and p-GSK3βexpression was up-regulated after NRK-C intervention,indicating that NRK-C might exert a potential antidiabetic effect by modulating the PI3K/AKT signaling pathway.CONCLUSION All these results suggested that NRK-C might improve T2DM and had the potential to be used as an adjunctive therapy.展开更多
Objective To evaluate the expression pattern of non-SMC condensin II complex subunit D3(NCAPD3)in hepatocellular carcinoma(HCC)tissues,assess its association with clinical characteristics,and explore the effects of NC...Objective To evaluate the expression pattern of non-SMC condensin II complex subunit D3(NCAPD3)in hepatocellular carcinoma(HCC)tissues,assess its association with clinical characteristics,and explore the effects of NCAPD3 on HCC cells and the potential underlying mechanisms.Methods NCAPD3 expression in HCC tumors and adjacent noncancerous tissues was quantified via quantitative PCR.Patients were divided into high-and low-expression groups on the basis of NCAPD3 levels,and associations with clinical parameters were assessed.The effects of NCAPD3 knockdown and the phosphatidylinositol-3-kinase(PI3K)agonist Y-P 740 on cell functions were examined via cell proliferation,Transwell migration,and invasion assays.Differentially expressed genes following NCAPD3 knockdown in SMMC-7721 cells were identified via mRNA sequencing.Western blotting was performed to measure NCAPD3,AKT serine/threonine kinase 1(AKT1),and phosphorylated AKT1 levels.Results NCAPD3 mRNA expression was notably upregulated in HCC tissues as compared with that in adjacent noncancer tissues.A positive correlation was observed between NCAPD3 expression and both lymphatic and distant metastases in patients with HCC.NCAPD3 knockdown reduced the proliferation and metastasis of SMMC-7721 and Huh-7 cells.mRNA sequencing revealed 140 downregulated genes and 125 upregulated genes.Further validation experiments confirmed that NCAPD3 modulated the PI3K-AKT signalling pathway and that the PI3K agonist Y-P 740 counteracted the effects of NCAPD3 knockdown.Conclusions Elevated NCAPD3 expression was strongly correlated with HCC metastasis.NCAPD3 inhibition impedes HCC cell growth and metastatic potential by suppressing the PI3K–AKT signalling pathway.展开更多
BACKGROUND MEX3A is a member of the human homologous gene MEX-3 family.It has been shown to promote cell proliferation and migration in various cancers,indicating its potential clinical significance.However,the role o...BACKGROUND MEX3A is a member of the human homologous gene MEX-3 family.It has been shown to promote cell proliferation and migration in various cancers,indicating its potential clinical significance.However,the role of MEX3A in hepatocellular carcinoma(HCC)remains largely unexplored,with limited reports available in the literature.AIM To investigate expression and clinical significance of MEX3A in HCC and explore its potential role in tumor progression.METHODS We analyzed MEX3A mRNA expression in HCC and adjacent tissues using data from The Cancer Genome Atlas(TCGA).The correlation between MEX3A expression and overall survival(OS)was evaluated.Immunohistochemistry was performed on HCC surgical specimens to validate MEX3A expression and its association with clinical parameters,including hepatitis B virus(HBV)positivity,tumor differentiation and tumor size.Additionally,MEX3A knockdown HCC cell lines were constructed to explore the biological functions of MEX3A.Cell prolif-eration was assessed using cell counting kit-8 and clone formation assays,while cell cycle progression was analyzed by flow cytometry.The effects of MEX3A on the Wnt/β-catenin signaling pathway were examined by western blotting and immunofluorescence.Cell migration was evaluated using scratch and Transwell assays.Finally,the role of the transcription factor RORA in mediating MEX3A effects was explored by silencing RORA and analyzing its impact on cell proliferation and protein expression.RESULTS TCGA data analysis revealed that MEX3A mRNA expression was significantly higher in HCC tissues compared to adjacent tissues.Higher MEX3A expression was associated with poorer OS.These findings were validated in HCC surgical specimens.Immunohistochemistry confirmed elevated MEX3A expression in HCC tissues and showed positive correlations with Ki-67 and vimentin levels.MEX3A expression was closely related to HBV positivity,tumor differentiation and tumor size.Mechanistic studies demonstrated that MEX3A knockdown inhibited cell proliferation and cell cycle progression,as shown by reduced expression ofβ-catenin,c-Myc and cyclin D1.Additionally,MEX3A knockdown inhibited the nuclear entry ofβ-catenin,thereby suppressing the activation of downstream oncogenic pathways.MEX3A depletion significantly reduced the migratory ability of HCC cells,likely through downregulation of the epithelial-mesenchymal transition pathway.Transcription factor analysis identified RORA as a potential mediator of MEX3A effects.Silencing RORA antagonized the effects of MEX3A on cell proliferation and the expression ofβ-catenin,c-Myc and cyclin D1.CONCLUSION MEX3A promotes cell proliferation in HCC by regulating the RORA/β-catenin pathway.Our findings suggest that MEX3A could serve as a prognostic marker and therapeutic target for HCC.展开更多
T-2 toxin,an omnipresent environmental contaminant,poses a serious risk to the health of humans and animals due to its pronounced cardiotoxicity.This study aimed to elucidate the molecular mechanism of cardiac tissue ...T-2 toxin,an omnipresent environmental contaminant,poses a serious risk to the health of humans and animals due to its pronounced cardiotoxicity.This study aimed to elucidate the molecular mechanism of cardiac tissue damage by T-2 toxin.Twenty-four male Sprague-Dawley rats were orally administered T-2 toxin through gavage for 12 weeks at the dose of 0,10,and 100 nanograms per gram body weight per day(ng/(g·day)),respectively.Morphological,pathological,and ultrastructural alterations in cardiac tissue were meticulously examined.Non-targeted metabolomics analysis was employed to analyze alterations in cardiac metabolites.The expression of the Sirt3/FoxO3α/MnSOD signaling pathway and the level of oxidative stress markers were detected.The results showed that exposure to T-2 toxin elicited myocardial tissue disorders,interstitial hemorrhage,capillary dilation,and fibrotic damage.Mitochondria were markedly impaired,including swelling,fusion,matrix degradation,and membrane damage.Metabonomics analysis unveiled that T-2 toxin could cause alterations in cardiacmetabolic profiles as well as in the Sirt3/FoxO3α/MnSOD signaling pathway.T-2 toxin could inhibit the expressions of the signaling pathway and elevate the level of oxidative stress.In conclusion,the T-2 toxin probably induces cardiac fibrotic impairment by affecting amino acid and choline metabolism as well as up-regulating oxidative stress mediated by the Sirt3/FoxO3α/MnSOD signaling pathway.This study is expected to provide targets for preventing and treating T-2 toxin-induced cardiac fibrotic injury.展开更多
BACKGROUND Liver cancer has a high incidence and mortality worldwide,especially in China.Herein,we investigated the therapeutic effect and mechanism of tetrahydrocurcumin against hepatocellular carcinoma(HCC),with a f...BACKGROUND Liver cancer has a high incidence and mortality worldwide,especially in China.Herein,we investigated the therapeutic effect and mechanism of tetrahydrocurcumin against hepatocellular carcinoma(HCC),with a focus on the of phosphoinositide 3-kinases(PI3K)/AKT signaling pathway.AIM To investigate the effects and mechanism of tetrahydrocurcumin in HCC cell lines HepG2 and Huh7.METHODS Using Metascape,we analyzed the potential targets of tetrahydrocurcumin in HCC.Molecular docking validation was performed using SYBYL2.0.Cell Counting Kit-8,wound healing,and transwell assays were performed to evaluate the effects of tetrahydrocurcumin on HepG2 and Huh7 cell migration,invasion,and apoptosis.The expression of PI3K/AKT signaling pathway-related proteins was detected by western blotting.RESULTS Network pharmacology and molecular docking showed that tetrahydrocurcumin has high binding affinity for phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha.In vitro experiments demonstrated that tetrahydrocurcumin suppressed the migration and invasion of liver cancer cells,promoted their apoptosis,and downregulated the expression of p-PI3K,p-AKT,and B cell leukemia/lymphoma 2,while upregulating caspase-3,p53,and B cell leukemia/lymphoma 2 associated X.CONCLUSION In summary,tetrahydrocurcumin suppresses PI3K/AKT signaling,promotes apoptosis,and prevents the migration and invasion of liver cancer cells.展开更多
Background Colitis caused by bacterial infection is a major global health challenge.Unfortunately,current treatment options are limited.We previously disclosed that L.reuteri SXDT-32 was enriched in the feces of an an...Background Colitis caused by bacterial infection is a major global health challenge.Unfortunately,current treatment options are limited.We previously disclosed that L.reuteri SXDT-32 was enriched in the feces of an ancient diarrhearesistant pig breed(Mashen pig)in China over 2500 years old.As diarrhea is often closely associated with intestinal inflammation,L.reuteri SXDT-32 was identified as a potential beneficial bacterium to prevent intestinal inflammation.However,the precise mechanisms involved remained unclear.Results Our tests showed that L.reuteri SXDT-32 alleviated colonic damage induced by pathogenic E.coli SKLAN202302 in weaned pigs by enhancing barrier integrity and inhibiting inflammation.The transcriptomics revealed that L.reuteri SXDT-32 protected against inflammatory injury by inhibiting the PI3K-AKT signaling pathway.Metabolite analysis indicated that the content of shikimic acid(SA)was substantially elevated in the colonic mucosa of L.reuteri SXDT-32-fed piglets(P<0.05).In addition,Liquid Chromatography-Mass Spectrometer(LC-MS)analysis showed significant increases in SA content in both the colonic chyme of L.reuteri SXDT-32-fed piglets and the supernatant of in vitro grown cultures of L.reuteri SXDT-32(P<0.05).Polymerase chain reaction(PCR)analysis identified gene aroE from L.reuteri SXDT-32,which is a key gene directly linked to SA synthesis,and elevated shikimate dehydrogenase(SD,encoded by aroE)was also detected in both L.reuteri SXDT-32 and the colonic mucosa of piglets fed L.reuteri SXDT-32(P<0.01).In vitro Caco-2 cell experiments demonstrated that SA,L.reuteri SXDT-32,and the supernatant from in vitro grown cultures of L.reuteri SXDT-32 exhibited comparable inhibitory effects on the PI3K-Akt pathway to those of the PI3K inhibitor LY294002.Conclusions L.reuteri SXDT-32 alleviated intestinal inflammation in piglets by producing SA that inhibits the PI3K-Akt pathway.This study provides an innovative approach for the treatment and prevention of colitis caused by bacterial infection.展开更多
BACKGROUND:Hepatic ischemia-reperfusion(I/R)injury is a major challenge in liver surgery and transplantation.Bromodomain protein 4(BRD4)has emerged as a promising target due to its role in oxidative stress and inflamm...BACKGROUND:Hepatic ischemia-reperfusion(I/R)injury is a major challenge in liver surgery and transplantation.Bromodomain protein 4(BRD4)has emerged as a promising target due to its role in oxidative stress and inflammation.JQ-1,a specific BRD4 inhibitor,has shown protective effects on organs suffering I/R injury.This study aims to investigate the expression of BRD4 in liver tissues after I/R injury and to explore its role in this process using JQ-1 both in vivo and in vitro.METHODS:Our study established a mouse model of hepatic I/R injury and investigated the protective effect of JQ-1.We compared the histological features,BRD4 expression,and liver enzyme levels between JQ-1-treated and untreated groups.Additionally,the antioxidant properties of JQ-1 were analyzed in RAW 264.7 cells by evaluating cytokine expression,NLRP3 inflammasome activity,and reactive oxygen species production.RESULTS:BRD4 was abundantly expressed in liver tissues after hepatic I/R injury,while JQ-1 treatment had antioxidant and hepatoprotective effects.JQ-1 also suppressed pro-inflammatory cytokine release in vitro.Furthermore,we clarified the mechanism by which JQ-1 enhances liver injury recovery through Kupffer cells by blocking the NOD-like receptor thermal protein domain-associated protein 3(NLRP3)/caspase-1 pathway.CONCLUSION:JQ-1 has potential as a pre-clinical emergency therapy for hepatic I/R injury.Its ability to inhibit BRD4 and modulate the inflammatory response in Kupffer cells offers a promising avenue for future clinical intervention.展开更多
BACKGROUND Macrophages are central to the orchestration of immune responses,inflammatory processes,and the pathogenesis of diabetic complications.The dynamic polarization of macrophages into M1 and M2 phenotypes criti...BACKGROUND Macrophages are central to the orchestration of immune responses,inflammatory processes,and the pathogenesis of diabetic complications.The dynamic polarization of macrophages into M1 and M2 phenotypes critically modulates inflammation and contributes to the progression of diabetic nephropathy.Sodiumglucose cotransporter 2 inhibitors such as dapagliflozin,which are acclaimed for their efficacy in diabetes management,may influence macrophage polarization,thereby ameliorating diabetic nephropathy.This investigation delves into these mechanistic pathways,aiming to elucidate novel therapeutic strategies for diabetes.AIM To investigate the inhibitory effect of dapagliflozin on macrophage M1 polarization and apoptosis and to explore its mechanism of action.METHODS We established a murine model of type 2 diabetes mellitus and harvested peritoneal macrophages following treatment with dapagliflozin.Concurrently,the human monocyte cell line cells were used for in vitro studies.Macrophage viability was assessed in a cell counting kit 8 assay,whereas apoptosis was evaluated by Annexin V/propidium iodide staining.Protein expression was examined through western blotting,and the expression levels of macrophage M1 surface immunosorbent assay,and quantitative real-time polymerase chain reaction analyses.RESULTS Dapagliflozin attenuated M1 macrophage polarization and mitigated apoptosis in the abdominal macrophages of diabetic mice,evidenced by the downregulation of proapoptotic genes(Caspase 3),inflammatory cytokines[interleukin(IL)-6,tumor necrosis factor-α,and IL-1β],and M1 surface markers(inducible nitric oxide synthase,and cluster of differentiation 86),as well as the upregulation of the antiapoptotic gene BCL2.Moreover,dapagliflozin suppressed the expression of proteins associated with the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway(PI3K,AKT,phosphorylated protein kinase B).These observations were corroborated in vitro,where we found that the modulatory effects of dapagliflozin were abrogated by 740Y-P,an activator of the PI3K/AKT signaling pathway.CONCLUSION Dapagliflozin attenuates the polarization of macrophages toward the M1 phenotype,thereby mitigating inflammation and promoting macrophage apoptosis.These effects are likely mediated through the inhibition of the PI3K/AKT signaling pathway.展开更多
Inflammation underlies many chronic diseases,and inflammatory bowel disease(IBD)is a condition characterized by long-term inflammation of the gut.Egg whites have been shown to contain many beneficial active substances...Inflammation underlies many chronic diseases,and inflammatory bowel disease(IBD)is a condition characterized by long-term inflammation of the gut.Egg whites have been shown to contain many beneficial active substances.Therefore,we obtained 2 peptides from salted egg white:Val-Val-His-Phe(VF-4)and Asp-Thr-Gln-Ala-Met-Pro-Phe-Arg(DR-8).The sodium dextran sulfate(DSS)-induced mice colitis model was used to evaluate its regulatory effect on colitis in vivo.The results showed that VF-4 and DR-8 improved the clinical symptoms of DSS-induced colitis,attenuated colon tissue damage,inhibited the activation of nuclear factor kappa-B(NF-κB)/mitogen-activated protein kinase(MAPK)/phosphoinositide 3-kinase-Akt(PI3K-AKT)signaling pathways,and inhibited the expression of inflammatory cytokines.16S rRNA gene sequencing showed that VF-4 and DR-8 administration increased the relative abundance of intestinal beneficial bacteria including Lactobacillus,Blautia,and down-regulated the relative abundance of inflammation-related bacteria including Acinetobacter,Lachnospiraceae_NK4A136_group,Klebsiella.Moreover,the degree of correlation between pro-inflammatory cytokines and microbiota was as follows:interleukin-6(IL-6)>tumor necrosis factor-α(TNF-α)>interleukin-1β(IL-1β)>interferon-γ(IFN-γ).In conclusion,this study suggests that salted egg white peptides VF-4 and DR-8 have a significant antiinflammatory effect in vivo.It also provides a strategy for the treatment of IBD and a new way for the highvalue utilization of salted egg white.展开更多
PI3K/AKT/mTOR signaling pathway is a key pathway of myocardial ischemia-reperfusion injury(MIRI).The mechanism of action is mainly oxidative stress,inflammatory response,calcium overload,ferroptosis,autophagy,and apop...PI3K/AKT/mTOR signaling pathway is a key pathway of myocardial ischemia-reperfusion injury(MIRI).The mechanism of action is mainly oxidative stress,inflammatory response,calcium overload,ferroptosis,autophagy,and apoptosis.MIRI belongs to the category of chest obstruction in traditional Chinese medicine,and its etiology and pathogenesis are mainly“Yang Wei Yin Xian.”Traditional Chinese medicine has the effect of multi-target and multi-component effect,and has played a significant role in the treatment of MIRI in recent years.At present,the monomers of traditional Chinese medicine mainly include saponins,flavonoids,alkaloids,terpenoids,and phenols,and the compounds mainly include Zhigancao Decoction,Zhenyuan Capsule,Jiawei Shenqibai Powder,Qili Qiangxin Capsule,Tongmai Yangxin Pill,Zhilong Huoxue Tongyu Capsule,Guizhi Tongluo Tablets,etc.This paper reviews the research on the improvement of MIRI by regulating PI3K/AKT/mTOR signaling pathway in recent years,and expounds the mechanism and advantages of traditional Chinese medicine in the treatment of MIRI.展开更多
Objectives:Colorectal cancer(CRC)is one of the most common cancers all over the world.The progression of CRC is associated with inflammation and disruptions in intestinal flora.3-O-Acetyl-11-keto-β-boswellic acid(AKB...Objectives:Colorectal cancer(CRC)is one of the most common cancers all over the world.The progression of CRC is associated with inflammation and disruptions in intestinal flora.3-O-Acetyl-11-keto-β-boswellic acid(AKBA)has been noted for its potent anti-inflammatory properties.However,the effect of AKBA on colon cancer caused by inflammation and its mechanism are not unclear.The study is to explore the effect of AKBA on CRC and its mechanism.Materials and Methods:Cell proliferation,(5-ethynyl-2′-deoxyuridine,EdU)-DNA synthesis assay and colony formation were used to assess the effect of AKBA on the proliferation of CRC cells.Flow cytometry was employed to analyze the cell cycle and apoptosis rate of cells treated with AKBA.RNA sequencing was done to explore the underlying mechanisms of AKBA.Western blot was used to assess the expression of key proteins in the nuclear factor kappa-B(NF-κB)signaling pathway after the treatment of AKBA.Real-time quantitative PCR(RT-qPCR),enzyme-linked immunosorbent assay(ELISA),and Meso Scale Discovery(MSD)assays were employed to check the anti-inflammation effects of AKBA on Lipopolysaccharide(LPS)-induced RAW264.7 cells and LPS-induced mouse model.Additionally,the Azoxymethane/Dextran sulfate sodium(AOM/DSS)-induced colitis-associated CRC model was used to evaluate the anti-CRC effect of AKBA.Gut microbiota profiling of fecal samples from CRC mice,both with and without AKBA treatment,was conducted through metagenomic sequencing analysis.Results:Our results showed that AKBA reduced the proliferation of HCT116 and SW620 cells,increased apoptosis of cells,and arrested the cell cycle at the G2/M phase.Results from RNA-seq showed that AKBA inhibited CRC by inhibiting the NF-κB signaling pathway and reducing cellular inflammation.Furthermore,AKBA reduced the levels of inflammatory cytokines,including tumor necrosis factor-α(TNF-α),Interferon-γ(IFN-γ),Interleukin-IL-12p70(IL-12p70),Interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)in both the spleen and serum of LPS-induced acute inflammation mice.Additionally,AKBA inhibited the development of AOM/DSS-induced colitis-associated colon cancer in mice and positively influenced gut microbiota.Conclusion:This study highlights the inhibitory effect of AKBA on colitis-associated CRC and reveals a novel aspect of its role in the remodeling of gut microbiota.These findings suggest that AKBA may be used as a potential therapeutic agent for CRC.展开更多
Objective To investigate the effect of electroacupuncture(EA)on microRNA(miRNA)expression spectrum and PI3K/Akt/mTOR signaling pathway in uterine tissue of rats with primary dysmenorrhea(PDM),and to explore the potent...Objective To investigate the effect of electroacupuncture(EA)on microRNA(miRNA)expression spectrum and PI3K/Akt/mTOR signaling pathway in uterine tissue of rats with primary dysmenorrhea(PDM),and to explore the potential mechanism of EA in the treatment of PDM.Methods Thirty female SD rats,weighted(200±20)g were randomly divided into control group,model group and EA group,10 rats in each group.By using subcutaneous injection of estradiol diphenhydrate combined with intraperitoneal injection of oxytocin,PDM models were established.Rats in the EA group received EA at“Sanyinjiao”(SP6)and“Guanyuan”(CV4)at dense waves and a frequency of 50 Hz,once a day,20 min each time,for 10 consecutive days.After the 10-day intervention,samples were collected and transmission electron microscopy was used to observe the ultrastructural changes of the cells in uterine tissue in each group.With RNA-seq method,the changes of miRNA expression spectrum in rat uterine tissue were detected.Bioinformatics analysis such as GO functional annotation and KEGG pathway was performed according to differentially expressed miRNAs.Differentially expressed miRNAs were verified by qRT-PCR.Endometrial stromal cells were selected as the target cells and transfected;and they were divided into control group,NC mimics group,mimic miR-144–3p group,NC inhibitor group and inhibitor miR-144–3p group.The apoptosis was determined by using flow cytometrydetect apoptosis,the miRNA and protein expression of PI3K/Akt/mTOR signaling pathway were detected by qRT-PCR and Western blot in each group separately.Results 1.Transmission electron microscope.(1)Control group:no obvious morphological changes in the uterine tissue.(2)Model group:fibroblasts in uterine tissue were irregular,the edema was presented in cellular cytoplasm,the nuclei were irregular and mitochondria swollen seriously;the rough endoplasmic reticulum was expanded moderately.(3)EA group:fibroblasts were spindle-shaped and pyknotic,the cytoplasm increased in electron density,the nuclei were slightly irregular and pyknotic,mitochondria were oval in shape,with little swelling and vacuolation;the rough endoplasmic reticulum was expanded slightly and retained,with a small amount of degranulation.2.Compared with the control group,there were 26 differentially expressed miRNAs in the uterine tissue of rats with PDM.After EA intervention,the expression of miR-144–3p was significantly up-regulated.GO functional analysis of differentially expressed miRNAs in PDM rats after EA showed that the biological functions involved calcium transmembrane transporter activity,mitogen-activated protein kinase binding,epithelial cell migration,tissue migration,etc.3.KEGG pathway analysis showed that PI3K/Akt signaling pathway,MAPK signaling pathway and calcium signaling pathway were enriched.Mimic miR-144-3p increased the apotosis of endometrial stromal cells,and decreased the mRNA and protein expression of PI3K,Akt,and mTOR(P<0.01).Conclusion EA can optimize the cell morphology in the uterine tissue of rats with PDM and affect the miRNA expression spectrum,which may be associated with the effect of EA for up-regulating miR-144–3p expression in endometrial stromal cells,suppressing PI3K/Akt/mTOR signaling pathway and causing apoptosis.展开更多
Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of th...Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of the neuroinflammation regulated by the FKBP prolyl isomerase 5(FKBP5)-IκB kinase alpha(IKKα)-nuclear factor kappa-B(NF-κB)-NOD-like receptor thermal protein domain-associated protein 3(NLRP3)signaling pathway in PTSD.Methods:The primary components of ADP,including ginsenosides Rg1 and Rb1,were quantified using ultra-performance liquid chromatography.Twelve C57BL/6 mice were allocated to control(D0)and experimental groups on days one,seven,and 14 of single prolonged stress(SPS).Eighteen C57BL/6 mice were allocated to control,SPS,and MCC950,an NLRP3 inhibitor(5 mg/kg)groups.Finally,24 C57BL/6 mice were allocated to control,SPS,paroxetine hydrochloride(PRX),or ADP(18.4 and 36.8 mg/kg)groups.Mice were administered MCC950,PRX,or ADP for 14 days.The open field test and elevated plus maze were used to evaluate anxiety-like behaviors,whereas fear memory extinction was evaluated using the fear memory test.Western blotting was employed to evaluate the expression levels of the FKBP5-IKKα-NF-κB-NLRP3 signaling pathway,tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β.The expression of FKBP5 and NLRP3 was further confirmed by immunofluorescence staining.Results:The amounts of ginsenosides Rg1 and Rb1 in ADP were(96.85±1.14)and(9.04±0.22)μg/g,respectively.Compared with the D0 group,the levels of the inflammatory cytokine proteins,TNF-α,IL-6,and IL-1β were elevated 1.33-to 1.51-fold and those of FKBP5-IKKα-NF-κB-NLRP3 signaling pathway were increased 1.16-to 1.41-fold in the hippocampus of the D14 group(P<0.05);the fluorescence intensity of FKBP5 and NLRP3 was also markedly increased(1.33-1.79-fold)in the hippocampus of the D14 group(P<0.5).Notably,injection of MCC950(5 mg/kg)reduced the levels of FKBP5-IKKα-NF-κB-NLRP3(0.80-0.88-fold)and inflammatory cytokines(0.74-0.83-fold),thereby improving the PTSD-like behaviors induced by SPS(P<0.05).In addition,ADP(36.8 g/kg)significantly improved PTSD-like behaviors and reduced levels of hippocampal inflammatory cytokines(0.70-0.79-fold)and FKBP5-IKKα-NF-κB-NLRP3(0.50-0.79-fold)(P<0.05)in SPS mice.Conclusion:The results suggest a potential therapeutic benefit of ADP in PTSD due to the inhibition of the FKBP5-IKKα-NF-κBNLRP3 signaling pathway.展开更多
BACKGROUND The interleukin-17(IL-17)mediated aberrant immune-inflammatory response plays a paramount role in ulcerative colitis(UC).γδT17 cells are one of the critical sources of IL-17,but the role they play in UC r...BACKGROUND The interleukin-17(IL-17)mediated aberrant immune-inflammatory response plays a paramount role in ulcerative colitis(UC).γδT17 cells are one of the critical sources of IL-17,but the role they play in UC remains under debate.AIM To clarify the role ofγδT17 cells in patients with mild-to-moderate UC.METHODS A single-centre observational pragmatic study was conducted on patients with UC who attended the outpatient and inpatient departments of Xiyuan Hospital of the China Academy of Traditional Chinese Medicine from September 2020 to December 2022.The research population consisted of two groups of adult patients.The first group consisted of healthy volunteers with no significant abnormalities on colonoscopy,and the other group consisted of patients with mild-to-moderate ulcerative colitis.Serum samples from healthy volunteers and patients with UC were collected for the detection of relevant inflammatory factors.Moreover,five colon mucosa samples were randomly selected from each group for testing and analyses.RESULTS An increased number ofγδT17 cells and hyperactivation of the NLR family pyrin domain containing 3/IL-1βsignaling pathway were observed in colonic mucosal tissues from patients with UC.CONCLUSION Hyperactivation of the NLR family pyrin domain containing 3/IL-1βsignaling pathway promotes the activation ofγδT17 cells in colonic mucosal tissues of patients with UC.展开更多
基金supported by the Program for Changjiang Scholars of the Ministry of Education of the People’s Republic of China (No. T2016088)the National Natural Science Foundation for Distinguished Young Scholars (No. 81725021)+4 种基金the National Key R&D Program of China (No. 2021YFA0910500)the Science and Technology Major Project of Hubei Province (No.2021ACA012)the Innovative Research Groups of the National Natural Science Foundation of China (No. 81721005)the Academic Frontier Youth Team of HUST (No. 2017QYTD19)the Fundamental Research Funds for the Central Universities (No.2172019kfy XJJS166)
文摘Cancer represents a significant disease that profoundly impacts human health and longevity.Projections indicate a 47%increase in the global cancer burden by 2040 compared to 2020,accompanied by a further rise in the associated economic burden.Consequently,there is an urgent need to discover and develop new alternative drugs to mitigate the global impact of cancer.Natural products(NPs)play a crucial role in the identification and development of anticancer therapeutics.This study identified ustusolate E(UE)and its analog 11α-hydroxy-ustusolate E(HUE)from strain Aspergillus calidoustus TJ403-EL05,and examined their antitumor activities and mechanisms of action.The findings demonstrate that both compounds significantly inhibited the proliferation and colony formation of AGS(human gastric cancer cells)and 786-O(human renal clear cell carcinoma cells),induced irreversible DNA damage,blocked the cell cycle at the G_(2)/M phase,and further induced apoptosis in tumor cells.To the best of the authors’knowledge,this is the first report on the anticancer effects of UE and HUE and their underlying mechanisms.The present study suggests that HUE and UE could serve as lead compounds for the development of novel anticancer drugs.
基金supported by the National Key Research and Development Program of China(No.2019YFC1712100)the National Natural Science Foundation of China(No.82105017)。
文摘Objective:The occurrence and development of atrial fibrillation(AF)are influenced by the autonomic nervous system and inflammation.Acupuncture is an effective treatment for AF.This study explored the protective effects of acupuncture in a rat model of paroxysmal AF and investigated its mechanisms.Methods:Male Sprague-Dawley rats(n=130)were randomly divided into blank control(Con),sham operation(Sham),AF,and acupuncture treatment(Acu)groups.A paroxysmal AF model was established by rapid atrial pacing through the jugular vein.Rats in the Acu group were immobilized to receive acupuncture treatment at Neiguan acupoint(PC6)for 20 min daily for seven days.The other groups were immobilized for the same duration over the treatment period but did not receive acupuncture.The AF induction rate,AF duration,cardiac electrophysiological parameters,and heart rate variability were evaluated by monitoring surface electrocardiogram and vagus nerve discharge signals.After the intervention,the rats were euthanized,and atrial morphology was assessed using haematoxylin and eosin staining.The expression of macrophage F4/80 antigen(F4/80)and cluster of differentiation(CD)86 in atrial myocardial tissue was detected using immunohistochemistry,immunofluorescence and flow cytometry.The expression levels or contents of interleukin(IL)-1β,IL-6,tumor necrosis factor-a(TNF-a),a7 nicotinic acetylcholine receptor(a7nAChR),phosphorylated Janus kinase 2(p-JAK2),and phosphorylated signal transducer and activator of transcription 3(p-STAT3)in atrial myocardial tissue were detected using Western blotting,reverse transcription-quantitative polymerase chain reaction,or enzyme-linked immunosorbent assay.The role of a7nAChR in acupuncture treatment was verified by intraperitoneal injection of the a7nAChR antagonist methyllycaconitine(MLA).Results:Compared with the AF group,acupuncture significantly reduced AF duration and induction rate,improved cardiac electrophysiology by enhancing vagus nerve activity and regulating autonomic balance.It also decreased the pro-inflammatory M1 macrophage proportion,alleviating myocardial injury and infiltration.MLA weakened acupuncture's electrophysiological improvement and anti-inflammatory effect.Results suggest that acupuncture triggers the a7nAChR-JAK2/STAT3 pathway and exerts cardioprotection via neuroimmune regulation.Conclusion:Acupuncture significantly reduced the AF induction rate,shortened AF duration,improved cardiac electrophysiological parameters,enhanced vagus nerve activity,and decreased the expression of pro-inflammatory M1 macrophages and inflammatory factors in rats with paroxysmal AF.
基金Mechanistic Investigation into the Extraction,Purification,and Anti-Esophageal Cancer Effects of Gallic Acid Derived from Rhodiola crenulata(YLUKLM2023001).
文摘Background:Gallic acid(GA),a plant-derived polyphenol,possesses diverse biological functions such as reducing inflammation and against tumors.Currently,the influence of GA on the resistance of esophageal squamous cell carcinoma(ESCC)cells to cisplatin(DDP)is not well understood.Methods:Cell counting kit-8 assay examined how GA affected KYSE30 and TE-1 cell viability.5-Ethynyl-2′-deoxyuridine and TdT-mediated dUTP Nick-End labeling staining detected cell proliferation and apoptosis.Clone formation assay,flow cytometry,Carboxyfluorescein diacetate succinimidyl ester fluorescent probes,and Transwell assay determined cell biological properties,and 2′,7′-Dichlorofluorescin diacetate(DCFH-DA)fluorescent probes detected oxidative stress levels.Signal transducer and activator of transcription 3(STAT3)/Notch pathway protein levels after GA and/or Interleukin-6(IL-6)intervention were examined through Western blot.Furthermore,a model for subcutaneous graft tumors was established in nude mice.Results:GA exerted suppressive effects on cell proliferation,and caused apoptosis of KYSE30 and TE-1 cells.IL-6 intervention activated the STAT3/Notch pathway and promoted the malignant biological properties of ESCC cells.In contrast,GA attenuated the effects of IL-6,while STAT3 or Notch inhibitor further enhanced the effects of GA,suggesting that GA inhibited the IL-6/STAT3/Notch pathway.Not only that,GA promoted oxidative stress and enhanced cell sensitivity to DDP both in vitro and in vivo.Conclusion:GA suppresses the malignant progression of ESCC and enhances cell sensitivity to DDP by hindering the IL-6/STAT3/Notch pathway.
基金Supported by Xi’an Science and Technology Plan Project,No.23YXYJ0162Shaanxi Province Traditional Chinese Medicine Research and Innovation Talent Plan Project,No.TZKN-CXRC-16+2 种基金Project of Shaanxi Administration of Traditional Chinese Medicine,No.SZYKJCYC-2025-JC-010Shaanxi Province Key Research and Development Plan Project-Social Development Field,No.S2025-YF-YBSF-0391the Science and Technology Innovation Cultivation Program of Longhua Hospital affiliated to Shanghai University of Chinese Medicine,No.YD202220。
文摘BACKGROUND Excessive endoplasmic reticulum(ER)stress in intestinal epithelial cells can lead to damage to the intestinal mucosal barrier,activate the signal transducer and activator of transcription 3(STAT3)/nuclear factor kappa B(NF-κB)signaling pathway,and exacerbate the inflammatory response,thus participating in the pathogenesis of ulcerative colitis(UC).Mesalazine is a commonly used drug in the clinical treatment of UC.However,further studies are needed to determine whether mesalazine regulates the ER stress of intestinal epithelial cells,downregulates the STAT3/NF-κB pathway to play a role in the treatment of UC.AIM To study the therapeutic effects of mesalazine on spontaneous colitis in interleukin-10(IL-10)-/-mice.METHODS The 24-week-old IL-10-/-mice with spontaneous colitis were divided into the model group and the 5-amino salicylic acid group.Littermates of wild-type mice of the same age group served as the control.There were eight mice in each group,four males and four females.The severity of symptoms of spontaneous colitis in IL-10-/-mice was assessed using disease activity index scores.On day 15,the mice were sacrificed.The colon length was measured,and the histopathological changes and ultrastructure of colonic epithelial cells were detected.The protein expressions of STAT3,p-STAT3,NF-κB,IκB,p-IκB,and glucoseregulated protein 78 were identified using Western blotting.The STAT3 and NF-κB mRNA expressions were identified using real-time polymerase chain reaction.The glucose-regulated protein 78 and C/EBP homologous protein expressions in colon sections were detected using immunofluorescence.RESULTS Mesalazine reduced the symptoms of spontaneous colitis in IL-10 knockout mice and the histopathological damage of colonic tissues,and alleviated the ER stress in epithelial cells of colitis mice.Western blotting and quantitative real-time polymerase chain reaction results showed that the STAT3/NF-κB pathway in the colon tissue of model mice was activated,suggesting that this pathway was involved in the pathogenesis of UC and might become a potential therapeutic target.Mesalazine could down-regulate the protein expressions of p-STAT3,NF-κB and p-IκB,and down-regulate the mRNA expression of STAT3 and NF-κB.CONCLUSION Mesalazine may play a protective role in UC by reducing ER stress by regulating the STAT3/NF-κB signaling pathway.
文摘AIM:To highlight the importance of microRNA(miRNA)-21-5p in directing the phosphatase and tensin homolog(PTEN)gene to control the phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)pathway in retinal pigment epithelial(RPE)cells in humans subjected to photodamage.METHODS:Human adult RPE cell line-19(ARPE-19)was cultured in vitro and randomly divided into control,damage,overexpression,negative,and PI3K/Akt blocker groups to establish a photodamage model of ARPE-19 cells.The models were subjected to 24h of light exposure,after which the corresponding indices were detected.The cell counting kit-8 assay quantified cell viability,while flow cytometry determined apoptosis rates.The miRNA-21 mimics and miRNA mimic NC were transfected into ARPE-19 cells using a transient transfection technique.Quantitative reverse transcription polymerase chain reaction(SYBR Green)and Western blotting analyzed expression levels of miRNA-21-5p,PTEN,p-PI3K/PI3K,p-mTOR/mTOR,and p-Akt/Akt.Statistical analyses comprised one-way analysis of variance and the Student-Newman-Keuls test for multiple group comparisons.RESULTS:The photodamage group demonstrated reduced cell survival rates than the control group(P<0.01).The overexpression group exhibited higher cell survival rates than the injury group(P<0.01).The negative group showed no difference in viability(P>0.05).The PI3K/Akt blocker group demonstrated lower cell viability,compared with the overexpression group(P<0.01).CONCLUSION:miRNA-21-5p significantly increases ARPE-19 cell survival after photodamage and inhibits lightinduced ARPE-19 cell apoptosis,suggesting that it may play a protective role in RPE by activating the PI3K/Akt/mTOR pathway while downregulating PTEN expression.
基金江苏省卫生健康委员会医学科研重点项目(K2023024)789 Outstanding Talent Program of SAHNMU(789ZYRC202090147)。
文摘Objective:To investigate the biological functions and molecular regulatory mechanisms of kinesin family member 11(KIF11)in colorectal cancer(CRC).Methods:The expression of KIF11 in CRC was examined by qRT⁃PCR and public databases.Functional assays(CCK⁃8,colony formation,EdU,and Transwell)were employed to evaluate KIF11’s roles in CRC progression.Western blot,RIP⁃qPCR,MeRIP⁃qPCR,and RNA stability assays were performed to elucidate the molecular mechanism of N6⁃methyladenosine(m6A)modification for KIF11.RNA sequencing(RNA⁃seq)and correlation analysis were used to examine the downstream mechanism of KIF11 regulation.Results:KIF11 was highly expressed in CRC and promoted CRC proliferation and migration.Mechanistically,methyltransferase⁃like 3(METTL3)/insulin like growth factor 2 mRNA binding protein 2(IGF2BP2)enhanced KIF11 mRNA stability and expression in an m6A⁃dependent way.Furthermore,by means of the PROM1/PI3K/AKT pathway,KIF11 facilitated the progression of CRC.Conclusion:The m6A modification of KIF11 by METTL3/IGF2BP2 contributes to CRC progression via the PI3K/AKT signaling pathway,highlighting its potential as a prognostic biomarker and therapeutic target.
文摘OBJECTIVE To explore hypoglycemic effect of 95%ethanol fraction of Nitraria roborowskii Kom(NRK-C)and its possible mechanism evaluated in the type 2 diabetes mellitus(T2DM)mice.METHODS The body weight,organ indices,blood glucose levels,serum biochemical indexes,as well as HE/PAS histopathological section were all analyzed to assess the hypoglycemic effect of NRK-C in T2DM mice induced by a high-fat diet(HFD)combined with six intraperitoneal injections of 35 mg·kg^(-1)of streptozotocin(STZ).The Western blotting and immunofluorescence were further applied to determine the regulatory effect of NRK-C on key signaling proteins.RESULTS The fasting blood glucose levels were significantly reduced after 7 weeks of administration of NRK-C.In addition,NRK-C could also significantly improve glucose tolerance,hepatic glycogen levels,and lipid levels(total cholesterol,triglyceride,low density lipoprotein and high density lipoprotein),and significantly reduced insulin resistance of diabetic mice,which played an important role in the antidiabetic effects.Further mechanism research demonstrated that phosphorylated PI3K expression was up-regulated and p-GSK3βexpression was up-regulated after NRK-C intervention,indicating that NRK-C might exert a potential antidiabetic effect by modulating the PI3K/AKT signaling pathway.CONCLUSION All these results suggested that NRK-C might improve T2DM and had the potential to be used as an adjunctive therapy.
基金supported by grants from Guangxi Nanning Qingxiu District Key Research and Development Program of Science and Technology Plan(no.2020050)Guangxi Medical and Health Appropriate Technology Development,Promotion and Application Project(no.S2021097)+1 种基金Guangxi Key Research and Development Program(no.AB22080064)Guangxi Natural Science Foundation(no.2017GXNSFAA198126).
文摘Objective To evaluate the expression pattern of non-SMC condensin II complex subunit D3(NCAPD3)in hepatocellular carcinoma(HCC)tissues,assess its association with clinical characteristics,and explore the effects of NCAPD3 on HCC cells and the potential underlying mechanisms.Methods NCAPD3 expression in HCC tumors and adjacent noncancerous tissues was quantified via quantitative PCR.Patients were divided into high-and low-expression groups on the basis of NCAPD3 levels,and associations with clinical parameters were assessed.The effects of NCAPD3 knockdown and the phosphatidylinositol-3-kinase(PI3K)agonist Y-P 740 on cell functions were examined via cell proliferation,Transwell migration,and invasion assays.Differentially expressed genes following NCAPD3 knockdown in SMMC-7721 cells were identified via mRNA sequencing.Western blotting was performed to measure NCAPD3,AKT serine/threonine kinase 1(AKT1),and phosphorylated AKT1 levels.Results NCAPD3 mRNA expression was notably upregulated in HCC tissues as compared with that in adjacent noncancer tissues.A positive correlation was observed between NCAPD3 expression and both lymphatic and distant metastases in patients with HCC.NCAPD3 knockdown reduced the proliferation and metastasis of SMMC-7721 and Huh-7 cells.mRNA sequencing revealed 140 downregulated genes and 125 upregulated genes.Further validation experiments confirmed that NCAPD3 modulated the PI3K-AKT signalling pathway and that the PI3K agonist Y-P 740 counteracted the effects of NCAPD3 knockdown.Conclusions Elevated NCAPD3 expression was strongly correlated with HCC metastasis.NCAPD3 inhibition impedes HCC cell growth and metastatic potential by suppressing the PI3K–AKT signalling pathway.
基金Supported by Suzhou Municipal Science and Technology Bureau,No.SYS2020081.
文摘BACKGROUND MEX3A is a member of the human homologous gene MEX-3 family.It has been shown to promote cell proliferation and migration in various cancers,indicating its potential clinical significance.However,the role of MEX3A in hepatocellular carcinoma(HCC)remains largely unexplored,with limited reports available in the literature.AIM To investigate expression and clinical significance of MEX3A in HCC and explore its potential role in tumor progression.METHODS We analyzed MEX3A mRNA expression in HCC and adjacent tissues using data from The Cancer Genome Atlas(TCGA).The correlation between MEX3A expression and overall survival(OS)was evaluated.Immunohistochemistry was performed on HCC surgical specimens to validate MEX3A expression and its association with clinical parameters,including hepatitis B virus(HBV)positivity,tumor differentiation and tumor size.Additionally,MEX3A knockdown HCC cell lines were constructed to explore the biological functions of MEX3A.Cell prolif-eration was assessed using cell counting kit-8 and clone formation assays,while cell cycle progression was analyzed by flow cytometry.The effects of MEX3A on the Wnt/β-catenin signaling pathway were examined by western blotting and immunofluorescence.Cell migration was evaluated using scratch and Transwell assays.Finally,the role of the transcription factor RORA in mediating MEX3A effects was explored by silencing RORA and analyzing its impact on cell proliferation and protein expression.RESULTS TCGA data analysis revealed that MEX3A mRNA expression was significantly higher in HCC tissues compared to adjacent tissues.Higher MEX3A expression was associated with poorer OS.These findings were validated in HCC surgical specimens.Immunohistochemistry confirmed elevated MEX3A expression in HCC tissues and showed positive correlations with Ki-67 and vimentin levels.MEX3A expression was closely related to HBV positivity,tumor differentiation and tumor size.Mechanistic studies demonstrated that MEX3A knockdown inhibited cell proliferation and cell cycle progression,as shown by reduced expression ofβ-catenin,c-Myc and cyclin D1.Additionally,MEX3A knockdown inhibited the nuclear entry ofβ-catenin,thereby suppressing the activation of downstream oncogenic pathways.MEX3A depletion significantly reduced the migratory ability of HCC cells,likely through downregulation of the epithelial-mesenchymal transition pathway.Transcription factor analysis identified RORA as a potential mediator of MEX3A effects.Silencing RORA antagonized the effects of MEX3A on cell proliferation and the expression ofβ-catenin,c-Myc and cyclin D1.CONCLUSION MEX3A promotes cell proliferation in HCC by regulating the RORA/β-catenin pathway.Our findings suggest that MEX3A could serve as a prognostic marker and therapeutic target for HCC.
基金supported by the National Natural Science Foundation of China(No.81872567).
文摘T-2 toxin,an omnipresent environmental contaminant,poses a serious risk to the health of humans and animals due to its pronounced cardiotoxicity.This study aimed to elucidate the molecular mechanism of cardiac tissue damage by T-2 toxin.Twenty-four male Sprague-Dawley rats were orally administered T-2 toxin through gavage for 12 weeks at the dose of 0,10,and 100 nanograms per gram body weight per day(ng/(g·day)),respectively.Morphological,pathological,and ultrastructural alterations in cardiac tissue were meticulously examined.Non-targeted metabolomics analysis was employed to analyze alterations in cardiac metabolites.The expression of the Sirt3/FoxO3α/MnSOD signaling pathway and the level of oxidative stress markers were detected.The results showed that exposure to T-2 toxin elicited myocardial tissue disorders,interstitial hemorrhage,capillary dilation,and fibrotic damage.Mitochondria were markedly impaired,including swelling,fusion,matrix degradation,and membrane damage.Metabonomics analysis unveiled that T-2 toxin could cause alterations in cardiacmetabolic profiles as well as in the Sirt3/FoxO3α/MnSOD signaling pathway.T-2 toxin could inhibit the expressions of the signaling pathway and elevate the level of oxidative stress.In conclusion,the T-2 toxin probably induces cardiac fibrotic impairment by affecting amino acid and choline metabolism as well as up-regulating oxidative stress mediated by the Sirt3/FoxO3α/MnSOD signaling pathway.This study is expected to provide targets for preventing and treating T-2 toxin-induced cardiac fibrotic injury.
基金the Central Government Guidance Local Science and Technology Development Fund,No.2023JH6/100100021Liaoning Province Education Department Foundation of China,No.JYTMS20231393+2 种基金Scientific Research Project of Liaoning Province Education Department,No.SYYX2019015Science and Technology Fund of Shenyang Medical College,No.20171004,Shenyang Medical College Master’s Science and Technology Innovation Fund,No.Y20220509Liaoning Provincial Department of Education Scientific Research Project,No.LJ212410164032.
文摘BACKGROUND Liver cancer has a high incidence and mortality worldwide,especially in China.Herein,we investigated the therapeutic effect and mechanism of tetrahydrocurcumin against hepatocellular carcinoma(HCC),with a focus on the of phosphoinositide 3-kinases(PI3K)/AKT signaling pathway.AIM To investigate the effects and mechanism of tetrahydrocurcumin in HCC cell lines HepG2 and Huh7.METHODS Using Metascape,we analyzed the potential targets of tetrahydrocurcumin in HCC.Molecular docking validation was performed using SYBYL2.0.Cell Counting Kit-8,wound healing,and transwell assays were performed to evaluate the effects of tetrahydrocurcumin on HepG2 and Huh7 cell migration,invasion,and apoptosis.The expression of PI3K/AKT signaling pathway-related proteins was detected by western blotting.RESULTS Network pharmacology and molecular docking showed that tetrahydrocurcumin has high binding affinity for phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha.In vitro experiments demonstrated that tetrahydrocurcumin suppressed the migration and invasion of liver cancer cells,promoted their apoptosis,and downregulated the expression of p-PI3K,p-AKT,and B cell leukemia/lymphoma 2,while upregulating caspase-3,p53,and B cell leukemia/lymphoma 2 associated X.CONCLUSION In summary,tetrahydrocurcumin suppresses PI3K/AKT signaling,promotes apoptosis,and prevents the migration and invasion of liver cancer cells.
基金supported by National Natural Science Foundation of China(U22 A20515)National High-Level Talents Special Support Program of China(2021-WR-01)Agricultural Science and Technology Innovation Program(ASTIPIAS07).
文摘Background Colitis caused by bacterial infection is a major global health challenge.Unfortunately,current treatment options are limited.We previously disclosed that L.reuteri SXDT-32 was enriched in the feces of an ancient diarrhearesistant pig breed(Mashen pig)in China over 2500 years old.As diarrhea is often closely associated with intestinal inflammation,L.reuteri SXDT-32 was identified as a potential beneficial bacterium to prevent intestinal inflammation.However,the precise mechanisms involved remained unclear.Results Our tests showed that L.reuteri SXDT-32 alleviated colonic damage induced by pathogenic E.coli SKLAN202302 in weaned pigs by enhancing barrier integrity and inhibiting inflammation.The transcriptomics revealed that L.reuteri SXDT-32 protected against inflammatory injury by inhibiting the PI3K-AKT signaling pathway.Metabolite analysis indicated that the content of shikimic acid(SA)was substantially elevated in the colonic mucosa of L.reuteri SXDT-32-fed piglets(P<0.05).In addition,Liquid Chromatography-Mass Spectrometer(LC-MS)analysis showed significant increases in SA content in both the colonic chyme of L.reuteri SXDT-32-fed piglets and the supernatant of in vitro grown cultures of L.reuteri SXDT-32(P<0.05).Polymerase chain reaction(PCR)analysis identified gene aroE from L.reuteri SXDT-32,which is a key gene directly linked to SA synthesis,and elevated shikimate dehydrogenase(SD,encoded by aroE)was also detected in both L.reuteri SXDT-32 and the colonic mucosa of piglets fed L.reuteri SXDT-32(P<0.01).In vitro Caco-2 cell experiments demonstrated that SA,L.reuteri SXDT-32,and the supernatant from in vitro grown cultures of L.reuteri SXDT-32 exhibited comparable inhibitory effects on the PI3K-Akt pathway to those of the PI3K inhibitor LY294002.Conclusions L.reuteri SXDT-32 alleviated intestinal inflammation in piglets by producing SA that inhibits the PI3K-Akt pathway.This study provides an innovative approach for the treatment and prevention of colitis caused by bacterial infection.
文摘BACKGROUND:Hepatic ischemia-reperfusion(I/R)injury is a major challenge in liver surgery and transplantation.Bromodomain protein 4(BRD4)has emerged as a promising target due to its role in oxidative stress and inflammation.JQ-1,a specific BRD4 inhibitor,has shown protective effects on organs suffering I/R injury.This study aims to investigate the expression of BRD4 in liver tissues after I/R injury and to explore its role in this process using JQ-1 both in vivo and in vitro.METHODS:Our study established a mouse model of hepatic I/R injury and investigated the protective effect of JQ-1.We compared the histological features,BRD4 expression,and liver enzyme levels between JQ-1-treated and untreated groups.Additionally,the antioxidant properties of JQ-1 were analyzed in RAW 264.7 cells by evaluating cytokine expression,NLRP3 inflammasome activity,and reactive oxygen species production.RESULTS:BRD4 was abundantly expressed in liver tissues after hepatic I/R injury,while JQ-1 treatment had antioxidant and hepatoprotective effects.JQ-1 also suppressed pro-inflammatory cytokine release in vitro.Furthermore,we clarified the mechanism by which JQ-1 enhances liver injury recovery through Kupffer cells by blocking the NOD-like receptor thermal protein domain-associated protein 3(NLRP3)/caspase-1 pathway.CONCLUSION:JQ-1 has potential as a pre-clinical emergency therapy for hepatic I/R injury.Its ability to inhibit BRD4 and modulate the inflammatory response in Kupffer cells offers a promising avenue for future clinical intervention.
基金Natural Science Foundation of Anhui Province,No.2208085MH216Major Natural Science and Technology Project of Bengbu Medical College,No.2020byfy004Scientific Research Program of Anhui Provincial Health Commission,No.AHWJ2023BAc10028.
文摘BACKGROUND Macrophages are central to the orchestration of immune responses,inflammatory processes,and the pathogenesis of diabetic complications.The dynamic polarization of macrophages into M1 and M2 phenotypes critically modulates inflammation and contributes to the progression of diabetic nephropathy.Sodiumglucose cotransporter 2 inhibitors such as dapagliflozin,which are acclaimed for their efficacy in diabetes management,may influence macrophage polarization,thereby ameliorating diabetic nephropathy.This investigation delves into these mechanistic pathways,aiming to elucidate novel therapeutic strategies for diabetes.AIM To investigate the inhibitory effect of dapagliflozin on macrophage M1 polarization and apoptosis and to explore its mechanism of action.METHODS We established a murine model of type 2 diabetes mellitus and harvested peritoneal macrophages following treatment with dapagliflozin.Concurrently,the human monocyte cell line cells were used for in vitro studies.Macrophage viability was assessed in a cell counting kit 8 assay,whereas apoptosis was evaluated by Annexin V/propidium iodide staining.Protein expression was examined through western blotting,and the expression levels of macrophage M1 surface immunosorbent assay,and quantitative real-time polymerase chain reaction analyses.RESULTS Dapagliflozin attenuated M1 macrophage polarization and mitigated apoptosis in the abdominal macrophages of diabetic mice,evidenced by the downregulation of proapoptotic genes(Caspase 3),inflammatory cytokines[interleukin(IL)-6,tumor necrosis factor-α,and IL-1β],and M1 surface markers(inducible nitric oxide synthase,and cluster of differentiation 86),as well as the upregulation of the antiapoptotic gene BCL2.Moreover,dapagliflozin suppressed the expression of proteins associated with the phosphoinositide 3-kinase(PI3K)/protein kinase B(AKT)signaling pathway(PI3K,AKT,phosphorylated protein kinase B).These observations were corroborated in vitro,where we found that the modulatory effects of dapagliflozin were abrogated by 740Y-P,an activator of the PI3K/AKT signaling pathway.CONCLUSION Dapagliflozin attenuates the polarization of macrophages toward the M1 phenotype,thereby mitigating inflammation and promoting macrophage apoptosis.These effects are likely mediated through the inhibition of the PI3K/AKT signaling pathway.
基金the financial support provided by the Project of Jiangxi Provincial Department of Education,China(GJJ200433)。
文摘Inflammation underlies many chronic diseases,and inflammatory bowel disease(IBD)is a condition characterized by long-term inflammation of the gut.Egg whites have been shown to contain many beneficial active substances.Therefore,we obtained 2 peptides from salted egg white:Val-Val-His-Phe(VF-4)and Asp-Thr-Gln-Ala-Met-Pro-Phe-Arg(DR-8).The sodium dextran sulfate(DSS)-induced mice colitis model was used to evaluate its regulatory effect on colitis in vivo.The results showed that VF-4 and DR-8 improved the clinical symptoms of DSS-induced colitis,attenuated colon tissue damage,inhibited the activation of nuclear factor kappa-B(NF-κB)/mitogen-activated protein kinase(MAPK)/phosphoinositide 3-kinase-Akt(PI3K-AKT)signaling pathways,and inhibited the expression of inflammatory cytokines.16S rRNA gene sequencing showed that VF-4 and DR-8 administration increased the relative abundance of intestinal beneficial bacteria including Lactobacillus,Blautia,and down-regulated the relative abundance of inflammation-related bacteria including Acinetobacter,Lachnospiraceae_NK4A136_group,Klebsiella.Moreover,the degree of correlation between pro-inflammatory cytokines and microbiota was as follows:interleukin-6(IL-6)>tumor necrosis factor-α(TNF-α)>interleukin-1β(IL-1β)>interferon-γ(IFN-γ).In conclusion,this study suggests that salted egg white peptides VF-4 and DR-8 have a significant antiinflammatory effect in vivo.It also provides a strategy for the treatment of IBD and a new way for the highvalue utilization of salted egg white.
基金Natural Science Foundation of Guangxi(Grant No.2021JJD140147)。
文摘PI3K/AKT/mTOR signaling pathway is a key pathway of myocardial ischemia-reperfusion injury(MIRI).The mechanism of action is mainly oxidative stress,inflammatory response,calcium overload,ferroptosis,autophagy,and apoptosis.MIRI belongs to the category of chest obstruction in traditional Chinese medicine,and its etiology and pathogenesis are mainly“Yang Wei Yin Xian.”Traditional Chinese medicine has the effect of multi-target and multi-component effect,and has played a significant role in the treatment of MIRI in recent years.At present,the monomers of traditional Chinese medicine mainly include saponins,flavonoids,alkaloids,terpenoids,and phenols,and the compounds mainly include Zhigancao Decoction,Zhenyuan Capsule,Jiawei Shenqibai Powder,Qili Qiangxin Capsule,Tongmai Yangxin Pill,Zhilong Huoxue Tongyu Capsule,Guizhi Tongluo Tablets,etc.This paper reviews the research on the improvement of MIRI by regulating PI3K/AKT/mTOR signaling pathway in recent years,and expounds the mechanism and advantages of traditional Chinese medicine in the treatment of MIRI.
基金financially supported by Beijing Natural Science Foundation[7252202,25JL008,China]from Beijing Natural Science Foundation CommitteeThis work was also supported by CAMS Innovation Fund for Medical Sciences[2021-I2M-1-029,2023-12M-2-006,China]and[2021-I2M-1-028,China]from Chinese Academy of Medical Science&Peking Union Medical College(CAMS&PUMC)+1 种基金National Natural Science Foundation of China[81703536 and 22278443,China]from Natural Science Foundation CommitteeInternational Cooperation Project of Qinghai Province[2020-HZ-803,China]from Science and Technology Department of Qinghai Province.
文摘Objectives:Colorectal cancer(CRC)is one of the most common cancers all over the world.The progression of CRC is associated with inflammation and disruptions in intestinal flora.3-O-Acetyl-11-keto-β-boswellic acid(AKBA)has been noted for its potent anti-inflammatory properties.However,the effect of AKBA on colon cancer caused by inflammation and its mechanism are not unclear.The study is to explore the effect of AKBA on CRC and its mechanism.Materials and Methods:Cell proliferation,(5-ethynyl-2′-deoxyuridine,EdU)-DNA synthesis assay and colony formation were used to assess the effect of AKBA on the proliferation of CRC cells.Flow cytometry was employed to analyze the cell cycle and apoptosis rate of cells treated with AKBA.RNA sequencing was done to explore the underlying mechanisms of AKBA.Western blot was used to assess the expression of key proteins in the nuclear factor kappa-B(NF-κB)signaling pathway after the treatment of AKBA.Real-time quantitative PCR(RT-qPCR),enzyme-linked immunosorbent assay(ELISA),and Meso Scale Discovery(MSD)assays were employed to check the anti-inflammation effects of AKBA on Lipopolysaccharide(LPS)-induced RAW264.7 cells and LPS-induced mouse model.Additionally,the Azoxymethane/Dextran sulfate sodium(AOM/DSS)-induced colitis-associated CRC model was used to evaluate the anti-CRC effect of AKBA.Gut microbiota profiling of fecal samples from CRC mice,both with and without AKBA treatment,was conducted through metagenomic sequencing analysis.Results:Our results showed that AKBA reduced the proliferation of HCT116 and SW620 cells,increased apoptosis of cells,and arrested the cell cycle at the G2/M phase.Results from RNA-seq showed that AKBA inhibited CRC by inhibiting the NF-κB signaling pathway and reducing cellular inflammation.Furthermore,AKBA reduced the levels of inflammatory cytokines,including tumor necrosis factor-α(TNF-α),Interferon-γ(IFN-γ),Interleukin-IL-12p70(IL-12p70),Interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α)in both the spleen and serum of LPS-induced acute inflammation mice.Additionally,AKBA inhibited the development of AOM/DSS-induced colitis-associated colon cancer in mice and positively influenced gut microbiota.Conclusion:This study highlights the inhibitory effect of AKBA on colitis-associated CRC and reveals a novel aspect of its role in the remodeling of gut microbiota.These findings suggest that AKBA may be used as a potential therapeutic agent for CRC.
基金supported by the National Natural Science Foundation of China(no.82004490)Hunan Natural Science Foundation Youth Project:2021JJ40402the Innovation Platform Open Fund project of the Hunan Education Department(no.20K091).
文摘Objective To investigate the effect of electroacupuncture(EA)on microRNA(miRNA)expression spectrum and PI3K/Akt/mTOR signaling pathway in uterine tissue of rats with primary dysmenorrhea(PDM),and to explore the potential mechanism of EA in the treatment of PDM.Methods Thirty female SD rats,weighted(200±20)g were randomly divided into control group,model group and EA group,10 rats in each group.By using subcutaneous injection of estradiol diphenhydrate combined with intraperitoneal injection of oxytocin,PDM models were established.Rats in the EA group received EA at“Sanyinjiao”(SP6)and“Guanyuan”(CV4)at dense waves and a frequency of 50 Hz,once a day,20 min each time,for 10 consecutive days.After the 10-day intervention,samples were collected and transmission electron microscopy was used to observe the ultrastructural changes of the cells in uterine tissue in each group.With RNA-seq method,the changes of miRNA expression spectrum in rat uterine tissue were detected.Bioinformatics analysis such as GO functional annotation and KEGG pathway was performed according to differentially expressed miRNAs.Differentially expressed miRNAs were verified by qRT-PCR.Endometrial stromal cells were selected as the target cells and transfected;and they were divided into control group,NC mimics group,mimic miR-144–3p group,NC inhibitor group and inhibitor miR-144–3p group.The apoptosis was determined by using flow cytometrydetect apoptosis,the miRNA and protein expression of PI3K/Akt/mTOR signaling pathway were detected by qRT-PCR and Western blot in each group separately.Results 1.Transmission electron microscope.(1)Control group:no obvious morphological changes in the uterine tissue.(2)Model group:fibroblasts in uterine tissue were irregular,the edema was presented in cellular cytoplasm,the nuclei were irregular and mitochondria swollen seriously;the rough endoplasmic reticulum was expanded moderately.(3)EA group:fibroblasts were spindle-shaped and pyknotic,the cytoplasm increased in electron density,the nuclei were slightly irregular and pyknotic,mitochondria were oval in shape,with little swelling and vacuolation;the rough endoplasmic reticulum was expanded slightly and retained,with a small amount of degranulation.2.Compared with the control group,there were 26 differentially expressed miRNAs in the uterine tissue of rats with PDM.After EA intervention,the expression of miR-144–3p was significantly up-regulated.GO functional analysis of differentially expressed miRNAs in PDM rats after EA showed that the biological functions involved calcium transmembrane transporter activity,mitogen-activated protein kinase binding,epithelial cell migration,tissue migration,etc.3.KEGG pathway analysis showed that PI3K/Akt signaling pathway,MAPK signaling pathway and calcium signaling pathway were enriched.Mimic miR-144-3p increased the apotosis of endometrial stromal cells,and decreased the mRNA and protein expression of PI3K,Akt,and mTOR(P<0.01).Conclusion EA can optimize the cell morphology in the uterine tissue of rats with PDM and affect the miRNA expression spectrum,which may be associated with the effect of EA for up-regulating miR-144–3p expression in endometrial stromal cells,suppressing PI3K/Akt/mTOR signaling pathway and causing apoptosis.
基金supported by the National Natural Science Foundation of China(82404995,82404890)Research Funds of Center for Xin’an Medicine and Modernization of Traditional Chinese Medicine of IHM(2023CXMMTCM013)+3 种基金Scientific Research Program of Anhui Provincial Department of Education(2024AH051036,2024AH040137,2024AH051044)Excellent Funding for Academic and Scientific Research Activities for Academic and Technological Leaders in Anhui Province(2022D317)Key Research and Development Plan of Anhui Province(202104j07020004)Anhui University of Chinese Medicine Talent Support Program(DT2300000173).
文摘Objective:Anshen Dingzhi prescription(ADP)is an effective remedy for treating post-traumatic stress disorder(PTSD);however,the mechanism underlying its beneficial effects is unclear.This study explores the roles of the neuroinflammation regulated by the FKBP prolyl isomerase 5(FKBP5)-IκB kinase alpha(IKKα)-nuclear factor kappa-B(NF-κB)-NOD-like receptor thermal protein domain-associated protein 3(NLRP3)signaling pathway in PTSD.Methods:The primary components of ADP,including ginsenosides Rg1 and Rb1,were quantified using ultra-performance liquid chromatography.Twelve C57BL/6 mice were allocated to control(D0)and experimental groups on days one,seven,and 14 of single prolonged stress(SPS).Eighteen C57BL/6 mice were allocated to control,SPS,and MCC950,an NLRP3 inhibitor(5 mg/kg)groups.Finally,24 C57BL/6 mice were allocated to control,SPS,paroxetine hydrochloride(PRX),or ADP(18.4 and 36.8 mg/kg)groups.Mice were administered MCC950,PRX,or ADP for 14 days.The open field test and elevated plus maze were used to evaluate anxiety-like behaviors,whereas fear memory extinction was evaluated using the fear memory test.Western blotting was employed to evaluate the expression levels of the FKBP5-IKKα-NF-κB-NLRP3 signaling pathway,tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β.The expression of FKBP5 and NLRP3 was further confirmed by immunofluorescence staining.Results:The amounts of ginsenosides Rg1 and Rb1 in ADP were(96.85±1.14)and(9.04±0.22)μg/g,respectively.Compared with the D0 group,the levels of the inflammatory cytokine proteins,TNF-α,IL-6,and IL-1β were elevated 1.33-to 1.51-fold and those of FKBP5-IKKα-NF-κB-NLRP3 signaling pathway were increased 1.16-to 1.41-fold in the hippocampus of the D14 group(P<0.05);the fluorescence intensity of FKBP5 and NLRP3 was also markedly increased(1.33-1.79-fold)in the hippocampus of the D14 group(P<0.5).Notably,injection of MCC950(5 mg/kg)reduced the levels of FKBP5-IKKα-NF-κB-NLRP3(0.80-0.88-fold)and inflammatory cytokines(0.74-0.83-fold),thereby improving the PTSD-like behaviors induced by SPS(P<0.05).In addition,ADP(36.8 g/kg)significantly improved PTSD-like behaviors and reduced levels of hippocampal inflammatory cytokines(0.70-0.79-fold)and FKBP5-IKKα-NF-κB-NLRP3(0.50-0.79-fold)(P<0.05)in SPS mice.Conclusion:The results suggest a potential therapeutic benefit of ADP in PTSD due to the inhibition of the FKBP5-IKKα-NF-κBNLRP3 signaling pathway.
基金Supported by China Academy of Chinese Medical Sciences Outstanding Young Talents(Innovative)Cultivation Project,No.ZZ17-YQ-007Special Project of the National Natural Science Foundation of China,No.82341233.
文摘BACKGROUND The interleukin-17(IL-17)mediated aberrant immune-inflammatory response plays a paramount role in ulcerative colitis(UC).γδT17 cells are one of the critical sources of IL-17,but the role they play in UC remains under debate.AIM To clarify the role ofγδT17 cells in patients with mild-to-moderate UC.METHODS A single-centre observational pragmatic study was conducted on patients with UC who attended the outpatient and inpatient departments of Xiyuan Hospital of the China Academy of Traditional Chinese Medicine from September 2020 to December 2022.The research population consisted of two groups of adult patients.The first group consisted of healthy volunteers with no significant abnormalities on colonoscopy,and the other group consisted of patients with mild-to-moderate ulcerative colitis.Serum samples from healthy volunteers and patients with UC were collected for the detection of relevant inflammatory factors.Moreover,five colon mucosa samples were randomly selected from each group for testing and analyses.RESULTS An increased number ofγδT17 cells and hyperactivation of the NLR family pyrin domain containing 3/IL-1βsignaling pathway were observed in colonic mucosal tissues from patients with UC.CONCLUSION Hyperactivation of the NLR family pyrin domain containing 3/IL-1βsignaling pathway promotes the activation ofγδT17 cells in colonic mucosal tissues of patients with UC.
