Objective:To investigate the potential mechanism of Wendan decoction in obesity by screening target genes with promoter region methylation changes and constructing a multiple signaling pathways network based on promot...Objective:To investigate the potential mechanism of Wendan decoction in obesity by screening target genes with promoter region methylation changes and constructing a multiple signaling pathways network based on promoter methylation.Methods:The methylation degree of Itgad,Col8a1,Adra2b,Jund,Rab2a,Wnt8b,Fzd9,B4galt7,Pik3cd,Creb1,Stard8,and Mmp1 in the abdominal adipose tissue of obese rats was determined using the Agena MassARRAY system.Western blot was performed to assess protein expression levels.Target genes were identified based on the methylation degree in the promoter region and protein expression.Enrichment analysis of signaling pathways was conducted to identify relevant target genes and obtain a multiple signaling pathway network associated with obesity.Core and terminal effector molecules in the pathway networks were selected as research targets for reverse transcription-polymerase chain reaction(RT-PCR)analysis.Results:Four genes(Adra2b,Creb1,Itgad,and Pik3cd)showed a degree of promoter methylation consistent with their respective protein expression levels.Among them,Adra2b,Creb1,and Pik3cd expression increased,while that of Itgad decreased.Enrichment analysis revealed that Creb1 and Pik3cd were involved in 6 signaling pathways related to obesity:tumor necrosis factor(TNF)signaling pathway,growth hormone synthesis/secretion and action,adenosine 5'-monophosphate-activated protein kinase(AMPK)signaling pathway,relaxin signaling pathway,cyclic nucleotide(cAMP)signaling pathway,and phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway.Subsequently,a multiple signaling pathways network was constructed based on promoter methylation.Key molecules including protein kinase B(AKT),mechanistic target of rapamycin complex 1(mTORC1),and unc-51 like autophagy activating kinase 1(ULK1),as well as terminal effector molecules interleukin-1β(IL-1β),interleukin-6(IL-6),and chemokine(C-X-C motif)ligand 2(CXCL2)were selected as research targets.Wendan decoction decreased the expressions of AKT,mTORC1,IL-1β,IL-6,and CXCL2 while up-regulating ULK1 expression.Conclusion:The mechanism of Wendan decoction in preventing obesity involves the regulation of multiple signaling pathways through the control of Creb1 and Pik3cd gene promoter methylation.However,the associated multi-path gene regulation mechanism in preventing obesity is complex.Thus,further exploration is needed to elucidate the role of methylation changes in this mechanism.展开更多
Obesity,caused by excessive energy,leads to body weight gain and various diseases,including cognitive impairment.Current studies suggest that diet restriction such as optimal fasting and regular exercise are crucial f...Obesity,caused by excessive energy,leads to body weight gain and various diseases,including cognitive impairment.Current studies suggest that diet restriction such as optimal fasting and regular exercise are crucial for improving cognitive capacity.However,further exploration is needed to understand the specific mechanisms of high fat diet(HFD)-induced cognitive decline in obesity.In the present study,4-month-old mice were subjected to HFD feeding for 18 weeks,followed by aerobic exercise and high-intensity intermittent exercise,regular diet feeding,and intermittent fasting for 8 weeks,and then used to evaluate cognitive capacity,inflammation,compromised insulin signaling pathway,and apoptosis in hippocampal tissue,as well as AMPK/SIRT1 and TLR4 signal pathways.Obese mice revealed impaired cognitive capacity as compared with mice fed with regular diets.In contrast,aerobic exercise,high-intensity intermittent exercise,regular diet,and intermittent fasting could inhibit apoptosis caused by inflammation-mediated compromised insulin signaling pathway in hippocampal tissues through activating the AMPK/SIRT1 signal pathway and suppressing the TLR4 signal pathway,thereby rescuing the cognitive impairment of obese mice.Therefore,diet restriction and exercise interventions may play a positive role in reverting obesity-induced cognitive impairment.展开更多
Objective To observe influence of moxibustion at ＂Shenshu＂ （肾俞BL 23） and ＂Zusanli＂ （足三里 ST 36） on proteins expression of Ras, ERKI/2, and Raf in synovial tissues in rats with rheumatoid arthritis （RA） t...Objective To observe influence of moxibustion at ＂Shenshu＂ （肾俞BL 23） and ＂Zusanli＂ （足三里 ST 36） on proteins expression of Ras, ERKI/2, and Raf in synovial tissues in rats with rheumatoid arthritis （RA） to elucidate molecular mechanisms of moxibustion on treating RA. Methods Thirty Wistar rats were randomly divided into a normal control group （Group Cont）, a model group （Group Mod） and a moxibustion group （Group Mox） with 10 rats in each group. The model was established by using both environmental factors such as wind, cold and damp and a biological factor （Freund＇s complete adjuvant）. Suspended moxibustion was applied at bilateral BL 23 and ST 36 for 20 min, applied once daily for 15 days, starting from the 3rd day after modeling. Volumes of metaleg digiti pedis on the right side were measured before, after and 15 days after modeling. Protein expressions of Ras, ERK 1/2, p-ERKI/2 and Raf in synovium tissues were detected by western blot. Results After modeling, volumes of metaleg digiti pedis on the right side were significantly enlarged, compared with those of rats in Group Cont （P〈0.01）. After treatment by moxibustion for 15 days, the metatarsale girth of metaleg digiti pedis on the right side in Group Mox was significantly reduced, compared with that in Group Mod （P〈0.01）. Protein expressions of Ras, Raf, ERK1/2 and p-ERK1/2 in Group Mod were significantly increased, compared with those in Group Cont （P〈0.01） and Group Mox （P〈0.01）. Protein expressions of Ras, Raf, ERK1/2 and p-ERK1/2 in Group Mox were significantly lowered, compared with those in Group Mod （all P〈0.01）. Conclusions Moxibustion can suppress excessive protein expressions of Ras, Raf and ERK1/2 in synovium tissues of rats with experimental RA to inhibit abnormal activation of Ras-MAPK signal Pathway, which may be one of the molecular mechanisms of moxibustion in alleviating synovitis.展开更多
Objective To investigate effect of pinacidil, an ATP sensitive potassium channel （KATP） opener, on the neuronal apoptosis and its signaling transduction mechanism following focal cerebral ischemia-reperfusion in rat...Objective To investigate effect of pinacidil, an ATP sensitive potassium channel （KATP） opener, on the neuronal apoptosis and its signaling transduction mechanism following focal cerebral ischemia-reperfusion in rats. Methods One hundred male Wistar rats were randomly divided into four groups： A, sham-operated group; B, ischemia-reperfusion group; C, KATe opener treatment group; and D, KATe opener and blocker treatment group. The middle cerebral artery occlusion （MCAO） model was established by using the intraluminal suture occlusion method, neuronal apoptosis was determined by TUNEL staining, and expressions of caspase-8, caspase-9 and caspase-3 mRNA were detected by in situ hybridization. Results （1） The numbers of apoptotic neurons at 12 h, 24 h, 48 h, and 72 h were significantly less in group C than in groups B and D （P 〈 0.01 or P 〈 0.05）; and there was no difference between groups B and D at all time points （P 〉 0.05）. （2） The expressions of caspase-3 mRNA and caspase-8 mRNA at all times and the expressions of caspase-9 mRNA at 12 h, 24 h, 48 h, 72 h were significantly lower in group C than in groups B and D （P 〈 0.01 or P 〈 0.05）; and there were no differences between groups B and D at all time points （P 〉 0.05）. Conclusions KATP opener can significantly decrease the neuronal apoptosis and the expressions of caspase-3, caspase-8 and caspase-9 mRNAs following cerebral ischemiareperfusion. The neuronal apoptosis may be decreased by the inhibition of both mitochondrial and death-receptor signal pathways.展开更多
Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a nov...Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a novel miRNA implicated in porcine IMF adipogenesis was found, and its effect and regulatory mechanism were further explored with respect to intramuscular preadipocyte proliferation and differentiation.Results: By porcine adipose tissue miRNA sequencing analysis, we found that miR-146a-5p is a potential regulator of porcine IMF adipogenesis. Further studies showed that miR-146a-5p mimics inhibited porcine intramuscular preadipocyte proliferation and differentiation, while the miR-146a-5p inhibitor promoted cell proliferation and adipogenic differentiation. Mechanistically, miR-146a-5p suppressed cell proliferation by directly targeting SMAD family member 4(SMAD4) to attenuate TGF-β signaling. Moreover, miR-146a-5p inhibited the differentiation of intramuscular preadipocytes by targeting TNF receptor-associated factor 6(TRAF6) to weaken the AKT/mTORC1 signaling downstream of the TRAF6 pathway.Conclusions: MiR-146a-5p targets SMAD4 and TRAF6 to inhibit porcine intramuscular adipogenesis by attenuating TGF-β and AKT/mTORC1 signaling, respectively. These findings provide a novel miRNA biomarker for regulating intramuscular adipogenesis to promote pork quality.展开更多
OBJECTIVE： To observe the influence of Sijunzi decoction and Yupingfeng powder on the expression of the relevant DNAs of janus kinase （JAK）-signal transducer and activator of transcription （STAT） sig- nal pathway...OBJECTIVE： To observe the influence of Sijunzi decoction and Yupingfeng powder on the expression of the relevant DNAs of janus kinase （JAK）-signal transducer and activator of transcription （STAT） sig- nal pathway of the brain in spleen-deficiency mod- el rats. METHODS： Eighty male Wistar rats of sanitary degree were divided randomly into four groups： nor- mal group, model group, treatment group 1, treatment group 2. Besides the rats in the normal group, all the rats in other 3 groups were prepared as spleen deficiency model. The treatment group 1 were treated with Sijunzi decoction and the treat- ment group 2 were treated with Yupingfeng powder. After treatment for 6 weeks, perfusion was given and the brain was taken for detection of the ex- pression of the relevant DNAs of JAK-STAT signal pathwayofthe brain in SD rats bygene chip method. RESULTS： Spleen deficiency could lead to increase expression of JAKI, STATI and Interleukin 4 （IL-4） in the brain, but the decrease expression of Suppres- sor of cytokine signaling I （SOCSI）, prolactin recep- tor （PRLR） and binding protein 3 （GATA 3）. Sijunzi decoction could increase expression of STAT3, Pro- lactin （PRL） and GATA3, but decrease expression of JAKI, STAT, STAT4, Interleukin 10 receptor, alpha （ILIORA）, Coagulation factor II （F2）, PRLR, MAD homolog 3 （SMAD3） and IL-4. Yupingfeng powder could decrease expression of JAKI, STATI, STAT4, SOCS4_ predicted, Epidermal growth factor receptor （EGFR）, PRLR, High mobility group AT-hook I （HMGAI 0）, IL-4. CONCLUSION： Sijunzi decoction and Yupingfeng powder can improve immune function of the rat through influencing the genetic expression of JAK-STAT signal pathway.展开更多
Accumulating evidence indicates that endostatin inhibits fibrosis. However, the mechanism is yet to be clarified. The aim of this study is to evaluate the effect of endostatin on platelet-derived growth factor-BB （PD...Accumulating evidence indicates that endostatin inhibits fibrosis. However, the mechanism is yet to be clarified. The aim of this study is to evaluate the effect of endostatin on platelet-derived growth factor-BB （PDGF-BB）- or transforming growth factor β1 （TGF-β1）-induced fibrosis in cultured human skin fibroblasts, and to further examine the molecular mechanisms involved. Human dermal flbroblasts were cultured in Dulbecco＇s modified Eagle＇s medium （DMEM） and serum-starved for 48 h before treatment. Cells were grouped as follows： ＂PDGF-BB＂, ＂PDGF-BB＋ endostatin＂, ＂TGF-β1＂, ＂TGF-β1＋endostatin＂, ＂endostatin＂, and ＂blank control＂. The fibroblasts were stimulated with either TGF-β1 or PDGF-BB for 72 h in order to set up the fibrosis model in vitro. The cells were co-cultured with either TGF-β1 or PDGF-BB and endostatin and were used to check the inhibiting effect of endostatin. A blank control group and an endostatin group were used as negative control groups. The biomarkers of fibrosis, including the expression of collagen I, hydrroxyproline, and α-smooth muscle actin （a-SMA）, were evaluated using an enzyme-linked immune- sorbent assay （ELISA） and Western blot. The expression of phosphorylated PDGF receptor β （p-PDGFRβ）, PDGFRβ, phosphorylated extracellular signal-regulated kinase （p-ERK）, and ERK was detected using Western blot and im- munofiuorescent staining was used to explore the mechanisms. Both PDGF-BB and TGF-β1 significantly up-regulated the expression of collagen I, hydroxyproline, and a-SMA. Endostatin significantly attenuated both the PDGF-BB- and TGF-β1-induced over-expression of collagen I, hydroxyproline, and a-SMA. PDGF-BB and TGF-β1 both promoted the expression of PDGFR, ERK, and p-ERK. Endostatin inhibited the expression of PDGFR and p-ERK but did not affect the expression of total ERK. Endostatin inhibited hypertrophic scar by modulating the PDGFRI3/ERK pathway. En- dostatin could be a promising multi-target drug in future fibrosis therapy.展开更多
Objective:To investigate the role of oxidative stress in human renal tubular epithelial cells(HK-2)induced by high glucose and the underlying signal pathway in vitro.Methods:MYPT1,pro-caspase-3,PGC-1α,and Drpl protei...Objective:To investigate the role of oxidative stress in human renal tubular epithelial cells(HK-2)induced by high glucose and the underlying signal pathway in vitro.Methods:MYPT1,pro-caspase-3,PGC-1α,and Drpl protein expressions were measured by Western blot.MnSOD2,Drp1 and PGC-1αmRNA expressions were detected by real time PCR.Results:Results showed that high glucose significantly up-regulated the protein expressions of MYPT1,pro-caspase-3 and the mRNA expression of MnSOD2 in HK-2 cells;while Rho kinase inhibitor fasudil and ROCK1 siRNA inhibited protein expressions of pro-caspase-3 and the mRNA expression of MnSOD2 in HK-2 cells induced by high glucose.Importantly,fasudil and ROCK1 siRNA markedly inhibited the expressions of mitochondrial motor proteins Drp1 and mitochondrial gene PGC-la in HK-2 cell=s induced by high glucose.Conclusions:Our findings suggest that Rho kinase signal pathway is involved in mitochondrial oxidative damage and apoptosis in high glucose-induced renal tubular epithelial cells by regulating mitochondrial motor proteins Drp1 and mitochondrial gene PGC-1α.Targeting Rho kinase signal pathway might be a potential strategy for the treatment of diabetic nephropathy.展开更多
The panax notoginseng saponin(PNS) had been clinically used for the treatment of cardiovascular diseases and stroke in China.It had been demonstrated that PNS could protect cardiomyocytes from injury induced by ischem...The panax notoginseng saponin(PNS) had been clinically used for the treatment of cardiovascular diseases and stroke in China.It had been demonstrated that PNS could protect cardiomyocytes from injury induced by ischemi- a,but the underlying molecular mechanisms of this protective effect were still unclear.This study was aimed to investigate the protective effect and molecular mechanisms of PNS on apoptosis in H9c2 cells in vitro and rat myocardial ischemia injury model in vivo.Annexin-V/PI assay shew that PNS could protect H9c2 cells from apoptosis induced by serum, glucose and oxygen deprivation(SGOD) in a dose-dependent manner.However,the anti-apoptotic effect of PNS was reversed by LY294002,a specific PI3K inhibitor.This antiapoptotic effect of PNS was confirmed by JC-1,a specific probe of mitochondrial membrane potential staining.PNS could significantly increase phos-Akt in H9c2 cells by Western blot assays and its effect could be inhibited by LY294002.Furthermore,PNS could improve ischemic-induced left ventricular function as reflected by EF,LVDd and LVDs.PNS could also inhibited cellular apoptosis in myocardial tissues in ischemic rats by TUNEL assay.PNS administration also increased the expression of phos-Akt in rat ischemic myocardial tissues.These results suggested that PNS could protect myocardial cells from apoptosis induced by ischemia in vitro model and in vivo model through activating-PI3K/Akt signal pathway which may be meaningful for further understanding the molecular mechanisms of cardiac protection of PNS.And the results might be useful in treatment of myocardial ischemia in future.展开更多
AIM:To investigate the feasibility and mechanism of immune tolerance in allergic conjunctivitis.METHODS:The allergic conjunctivitis immune tolerance mice model was established by ragweed pollen(RW)and the related cyto...AIM:To investigate the feasibility and mechanism of immune tolerance in allergic conjunctivitis.METHODS:The allergic conjunctivitis immune tolerance mice model was established by ragweed pollen(RW)and the related cytokines were detected.The mice were divided into 9 groups and the maslinic acid(MA)or PBS were given for different group after modeling.The expression levels of chemokine ligand 5(CCL5)and P-65 in the conjunctival tissue were analyzed by immunohistochemistry,quantitative reverse transcription polymerase chain reaction(q RT-PCR)and Western blot.The percentage of interleukin-17(IL-17)and CD4+CD25+in the splenocyte supernatant was analyzed by flow cytometry.Fur thermore,the serum and splenocyte supernatant concentration of total-IgE,interleukin-10(IL-10),and IL-17 was analyzed by enzyme linked immune response(ELISA).RESULTS:After the model was established,symptoms of conjunctivitis were alleviated,the level of P-65,CCL5,IL-17,and total-IgE was raised,while the expression of IL-10,CD4+CD25+was decreased.This result fully demonstrated that a typical IL-17/regulatory-T-cells(Treg cells)imbalance and NF-κB activation.When the NF-κB signal pathway was suppressed,it showed that there was a further relief of conjunctivitis in mice.At the same time,the expression of total-IgE,IL-17,and CCL5 was decreased and the expression of anti-inflammatory factor(IL-10,CD4+CD25+)was increased.CONCLUSION:In the state of immune tolerance,symptoms of conjunctivitis in mice are alleviated,the Th-17 cells of allergic conjunctivitis mice are inhibited,and Treg cells activity is enhanced.展开更多
Hepatic fibrosis is a reversible pathological phenomenon in the early and middle stages,but but no satisfactory intervention drugs have been available so far.Recent studies have suggested that microcirculation disturb...Hepatic fibrosis is a reversible pathological phenomenon in the early and middle stages,but but no satisfactory intervention drugs have been available so far.Recent studies have suggested that microcirculation disturbance of liver is one of the important pathogenesis of chronic liver disease,the improvement of microcirculation is beneficial to the recovery of liver function and the delay of liver fibrosis.Hepatic stellate cells are the core cells of hepatic fibrosis,and also the most critical cells that affect the microcirculation of the liver.While TLR4/MyD88/NF-κB and TLR4/MyD88/MAPKs which are based on the action of hepatic stellate cells are two pathways that have very important influence on the inflammatory response of liver,the proliferation and apoptosis of hepatic stellate cells,and the secretion of fibrogenic cytokines.It was found that Plumbapin,the active ingredient of Guangxi specialty ethnic medicine,has the definite effect of promoting blood circulation and removing blood stasis and anti-hepatic fibrosis,but its mechanism is not clear.In this study,the research progress of the above problems was reviewed,and further research ideas were derived as follows:the pharmacological effect of Plumbapin on anti-hepatic fibrosis,promoting blood circulation and removing stasis was based on the influence of TLR4/MyD88/NF-κB and MAPKs signal pathway.展开更多
[Objectives]To explore the effect and possible mechanism of bergenin in relieving allergic rhinitis(AR)in mice.[Methods]50 C57/BL6 mice were randomly divided into blank group(n=10),model group(n=10)and high(100 mg/kg)...[Objectives]To explore the effect and possible mechanism of bergenin in relieving allergic rhinitis(AR)in mice.[Methods]50 C57/BL6 mice were randomly divided into blank group(n=10),model group(n=10)and high(100 mg/kg),medium(50 mg/kg)and low(25 mg/kg)dose bergenin groups with 10 mice in each group.Except for the blank group,the other mice were sensitized by basic ways combined with attack to replicate the AR model.From the 15th d of modeling(from the second d after the end of the basic modeling),the drug group was given bergenin orally for 15 d,and the blank group and model group were given the same volume of normal saline once a day.24 h after the last establishment of the model,the content of interleukin 4(IL-4),IL-6,TNF-αand IL-1βin nasal lavage fluid and serum of mice in each group was detected by ELISA.The expression of TLR-4,NF-κB and p-NF-κB in nasal mucosa of mice was detected by Western blot.[Results]Compared with the blank group,the content of inflammatory factors IL-4,IL-6,TNF-αand IL-1βin nasal lavage fluid and serum of model group was significantly increased,and the protein expression of TLR-4 and p-NF-κB was significantly increased.After the intervention of bergenin,the content of IL-4,IL-6,TNF-αand IL-1βin nasal lavage fluid and serum and TLR-4 and p-NF-κB protein in tissue was significantly inhibited in bergenin group.[Conclusions]Bergenin can effectively reduce allergic inflammation in AR model mice,and its mechanism may be related to inhibition of inflammation and down-regulation of TLR-4/NF-κB signal pathway.展开更多
Background To investigate the effects and mechanisms of cellular repressor of ElA stimulated genes (CREG) on endothelial cell(EC) migration.Methods vascular endothelial cells(VE),CREG overexpression VEs, CREG suppress...Background To investigate the effects and mechanisms of cellular repressor of ElA stimulated genes (CREG) on endothelial cell(EC) migration.Methods vascular endothelial cells(VE),CREG overexpression VEs, CREG suppression VEs and VEs transfected with CREG gene modified adenovirus(Ad-CREG) were cultured with dulbecco’s modified eagle’s medium contained 10%fetal calf serum. Western blot was used to detect the protein level of CREG and integrin-linked kinase(ILK) in the four kind ECs.Tran-swell migration model was applied to compare the migration cell number of the four kind ECs.Two kinds of ILK mutant plasmids;PCXN2-flag-ILK wt-IRES-GFP(wild-type ILK)and PCXN2-flag-ILK p-parvin-IRES-GFP(P-parvin-binding mutant) were used to transfect VS and VE respectively,then the two kind transfection ECs were named as VS-wtILK and VE-P -parvin which were selected by G418(600ng/ml)for 2 weeks;Transwell migration model was applied to compare migration capability before and after ILK plasmids transfecting VE and VS.Results Western blot analysis showed that CREG overexpression promoted ILK expression in ECs,on the contrary,ILK expression was down-regulated in CREG silent ECs(P【0.05).Further more,ILK expression was up-regulated obviously in VE transfected with Ad-CREG(P【 0.05);Transwell migration model showed that EC’s migration capability was positively correlated with the expression level of CREG in EC,that is,CREG overexpression induced VE migration and CREG silent suppressed VE migration, moreover,Ad-CREG transfecting VE showed better migration capability accompanied with CREG expression increase by transwell migration model(P【0.05).In order to know the relationship between ILK expression and cell migration,we obtained stable transfection cell strains of VS-wtILK and VE-Pparvin, transwell migration model demonstrated that VS-wtILK remarkably corrected the poor migration capability of VS(P【 0.01),butβ-parvin combining site mutation in ILK genes inhibited VE migration markedly(P【0.01).Conclusions ILKp -parvin signal pathway mediated vascular endothelial cell migration induced by CREG.展开更多
The melanoma differentiation-associated gene-7（mda-7）,IL-24,has the specific functions that induce cancer cell apoptosis without doing harm to normal cells. We systematically review the apoptotic signal pathways and...The melanoma differentiation-associated gene-7（mda-7）,IL-24,has the specific functions that induce cancer cell apoptosis without doing harm to normal cells. We systematically review the apoptotic signal pathways and their regulatory mechanisms induced by Ad.IL-24 and IL-24 in diverse cancer cells. IL-24 can participate in varied signal transduction pathways,including JAK,p38 MAPK,Wnt/β-catenin,JNK,ER stress and mitochondria-associated signal pathways. And we review five proteins interacting with IL-24,including Bip/GRP78,S1 R,PKR,Beclin1 and soluble clusterin,which are relative to the tumor-specific effect of IL-24. It is speculated that ER stress,G-protein pathways and MAPK signal pathways may be the primary upstream effectors which activate the sequential downstream mediators resulting in apoptosis induced by IL-24 in tumor cells. Experimental results also show that IL-24 sensitizes cancer cells and indirectly promotes apoptosis rather than functions as a direct apoptosis inducer itself.展开更多
Psoriasis is a complex skin disease and the pathogenesis of psoriasis is not clear.The purpose of this study is to identify the key driving genes and signal pathways involved in psoriasis and to predict the potential ...Psoriasis is a complex skin disease and the pathogenesis of psoriasis is not clear.The purpose of this study is to identify the key driving genes and signal pathways involved in psoriasis and to predict the potential miRNA,for further understanding the pathogenesis of psoriasis.Methods:Three gene expression profiling chips,including GSE67853,GSE78097,and GSE136757 with a total of 120 samples were collected and analyzed with R software.The protein-protein interaction network of differentially expressed genes was constructed with STRING database and Cytoscape.CIBERSORT was used to evaluate the infiltration of immune cells in psoriasis tissues,and the correlation between diagnostic markers and infiltrating immune cells was analyzed.Further,the key biomarkers were identified and the targeting miRNA of crucial genes was predicted.Results:A total of 201 differentially expressed genes(163 upregulated genes and 38 downregulated genes)were determined.CXCL1,CXCL2,and CXCL8,the critical biomarkers of psoriasis,were identified by different calculation methods.The potential critical signal pathway NOD-like receptor signaling pathway of psoriasis was explored by gene expression profiling chip gene enrichment analysis and differentially expressed gene enrichment analysis.Immune cell infiltration analysis found that CXCL1,CXCL2,CXCL8 was positively correlated with macrophages M1 and T cells CD4 memory activated and negatively correlated with macrophages M2 and mast cells resting.At the same time,through miRNA prediction,we found that hsa-miR-216a-3p and hsa-miR-6750-5p can be used as potential psoriasis targets.Conclusions:This research proposes a new comprehensive strategy to identify psoriasis’s potential biomarkers through cross-validation and significant scores of different calculation methods.In this research,we identified CXCL1,CXCL2,and CXCL8 as potential key biomarkers of psoriasis,and the NOD-like receptor signaling pathway is the critical signal pathway of psoriasis.Hsa-miR-216a-3p and hsa-miR-6750-5p can be used as potential psoriasis targets.展开更多
Objective: to explore the effect of micro-current stimulation on activation or inhibition of relevant signal pathways (MAPK, Wnt/β-catenin, NF-κB) in LDH model rats. Methods: a total of 180 healthy adult SD rats, ha...Objective: to explore the effect of micro-current stimulation on activation or inhibition of relevant signal pathways (MAPK, Wnt/β-catenin, NF-κB) in LDH model rats. Methods: a total of 180 healthy adult SD rats, half male and half female, weighing 250-280g, were selected for modeling. After successful modeling, NPCs cells of intervertebral disc were divided into stimulation group and non-stimulation group according to the intervention of low-frequency electronic pulse current stimulation. The contents of MAPK, Wnt/β-catenin, NF-κB signal pathway and LDH-related characteristic indicators in the two groups of rats were compared. Results: the relative expressions of p-p38MAPK, p-ERK1, p-ERK2 and p-JNK in the stimulation group were significantly lower than those in the non-stimulation group (p 0.05). The mechanical stimulation pain threshold of the stimulation group was significantly higher than that of the non-stimulation group, and the content of IL-1β and TNF-α was significantly lower than that of the non-stimulation group (P 0.05). There was no significant difference in IL-6 between the two groups. Conclusion: microcurrent stimulation can reduce the relative expression of p-p38MAPK, p-ERK1, p-ERK2, p-JNK protein in rat dorsal root ganglion, suggesting that it can inhibit ERK signaling pathway in rats with lumbar disc herniation.展开更多
OBJECTIVE To investigate the protective effect of salvianolic acid A(Sal A)on isoproterenol-induced myocardial infarction in mice and its possible mechanisms.METHODS The mice were subcutaneously injected with isopropr...OBJECTIVE To investigate the protective effect of salvianolic acid A(Sal A)on isoproterenol-induced myocardial infarction in mice and its possible mechanisms.METHODS The mice were subcutaneously injected with isopropranol(ISO 8 mg·kg-1)to induce myocardial infarction and evaluated the myocardial protective effect of Sal A from mortality rate,electrocardiogram(ECG),heart function,myocardial infarction index,serum myocardial enzymes and explored its possible mechanisms from inflammatory,antioxidant and cells apoptosis.RESULTS Sal A can dose-dependently enhanced the heart function of myocardial infarction mice,reduced the heart index,inhibited the myocardial enzyme leakage,showed obvious myocardial protection effects.ELISA results showed that Sal A can reduce the expression of myocardial inflammatory cytokines such as IL-6,TNF-α.Western blotting confirmed that Sal A can increase the expression of anti-apoptotic proteins Bcl-2,reduce the expression of apoptosis protein Bax,and raise the phosphorylation level of PI3K and Akt.CONCLUSION Sal A have displayed significant protective effect against isoproterenol-induced myocardial infarction and its mechanism may be related to increasing of PI3K/Akt signal pathway and inhibition of cell apoptosis and inflammatory reaction.展开更多
Heart failure(HF)is a kind of continuous development syndrome of cardiac insufficiency caused by various heart diseases.Not only does the prevalence continue to rise,but the mortality rate and readmission rate remain ...Heart failure(HF)is a kind of continuous development syndrome of cardiac insufficiency caused by various heart diseases.Not only does the prevalence continue to rise,but the mortality rate and readmission rate remain high.Heart failure is also the end-stage of cardiovascular disease and the main cause of death of patients,which seriously affects the health and quality of life of people all over the world.Ventricular remodeling plays a key role in the occurrence and development of heart failure.Therefore,by improving ventricular remodeling,it is of important research value to explore the intervention of traditional Chinese medicine in the development of heart failure.Studies have shown that the mediation of multiple signaling pathways can lead to progressive aggravation of ventricular remodeling,and experimental studies often confirm the therapeutic effects of traditional Chinese medicine.Traditional Chinese medicine usually achieves the therapeutic effect of heart failure through multiple targets and multiple approaches.In recent years,there have been more and more researches on the role and mechanism of Chinese medicine intervention in heart failure.However,it is concluded that Chinese medicine intervention has less influence on heart failure signal pathways.This article summarizes the understanding of Chinese medicine on heart failure and the five signal pathways related to Chinese medicine intervention in heart failure.The 5 signaling pathways in the world,namely transforming growth factor-β1(TGF-β1)/signal transduction protein(Smads)signaling pathway,Toll-like receptor(TLR)/nuclear transcription factor-κB(NF-κB)inflammation signaling pathway,Renin-angiotensinaldosterone(RAAS)system,phosphoinositide 3-kinase(PI3K)-serine/threonine protein kinase(AKT)signaling pathway and mitogen-activated protein kinase(MAPK)dependent signaling pathway.展开更多
Objective: To observe the effect of acupuncture on the expression of ERM/PI3K/Akt signaling pathway in the brain tissue around the infarct focus of ischemic stroke model rats. Methods: According to the block randomiza...Objective: To observe the effect of acupuncture on the expression of ERM/PI3K/Akt signaling pathway in the brain tissue around the infarct focus of ischemic stroke model rats. Methods: According to the block randomization method, 80 male SD rats were divided into normal group (n = 10), sham operation group (n = 24), model group (n = 23) and acupuncture group (n = 23). The normal group was bred routinely without any intervention. In the sham operation group, only the skin was cut to find and the nerves and blood vessels were stripped and sutured. The model group and the acupuncture group were sutured to prepare the MCAO model. After the model was successfully established, acupuncture of the acupuncture group rats was given once a day, and the needles were kept for 30 minutes each time. The rats in the sham operation group and the model group only imitated capture and return, and were treated with acupuncture for 2 consecutive courses, each course was 5 days, and the two courses were rested for 2 days. After that, Western blot was used to detect ERM, changes in the concentration of PI3K and Akt proteins. Results: Compared with the normal group, the expression levels of ERM, PI3K, and Akt proteins in the sham operation group were reduced. Compared with the sham operation group, the expression levels of ERM, PI3K, and Akt proteins in the model group were significantly down-regulated. Compared with the model group, the protein expression levels of ERM, PI3K and Akt were significantly up-regulated in the acupuncture group, and the difference was statistically significant (P < 0.05). Conclusion: Acupuncture can significantly up-regulate the expression of ERM, PI3K and Akt proteins, and its effect mechanism may be related to the transduction of ERM/PI3K/Akt signaling pathway in cells.展开更多
BACKGROUND Simulated microgravity environment can lead to gastrointestinal motility disturbance.The pathogenesis of gastrointestinal motility disorders is closely related to the stem cell factor(SCF)/c-kit signaling p...BACKGROUND Simulated microgravity environment can lead to gastrointestinal motility disturbance.The pathogenesis of gastrointestinal motility disorders is closely related to the stem cell factor(SCF)/c-kit signaling pathway associated with intestinal flora and Cajal stromal cells.Moreover,intestinal flora can also affect the regulation of SCF/c-kit signaling pathway,thus affecting the expression of Cajal stromal cells.Cajal cells are the pacemakers of gastrointestinal motility.AIM To investigate the effects of Bifidobacterium lactis(B.lactis)BLa80 on the intestinal flora of rats in simulated microgravity and on the gastrointestinal motility-related SCF/c-kit pathway.METHODS The internationally recognized tail suspension animal model was used to simulate the microgravity environment,and 30 rats were randomly divided into control group,tail suspension group and drug administration tail suspension group with 10 rats in each group for a total of 28 days.The tail group was given B.lactis BLa80 by intragastric administration,and the other two groups were given water intragastric administration,the concentration of intragastric administration was 0.1 g/mL,and each rat was 1 mL/day.Hematoxylin&eosin staining was used to observe the histopathological changes in each segment of the intestine of each group,and the expression levels of SCF,c-kit,extracellular signal-regulated kinase(ERK)and p-ERK in the gastric antrum of each group were detected by Western blotting and PCR.The fecal flora and mucosal flora of rats in each group were detected by 16S rRNA.RESULTS Simulated microgravity resulted in severe exfoliation of villi of duodenum,jejunum and ileum in rats,marked damage,increased space between villi,loose arrangement,shortened columnar epithelium of colon,less folds,narrower mucosal thickness,reduced goblet cell number and crypts,and significant improvement after probiotic intervention.Simulated microgravity reduced the expressions of SCF and c-kit,and increased the expressions of ERK and P-ERK in the gastric antrum of rats.However,after probiotic intervention,the expressions of SCF and ckit were increased,while the expressions of ERK and P-ERK were decreased,with statistical significance(P<0.05).In addition,simulated microgravity can reduce the operational taxonomic unit(OTU)of the overall intestinal flora of rats,B.lactis BLa80 can increase the OTU of rats,simulated microgravity can reduce the overall richness and diversity of stool flora of rats,increase the abundance of firmicutes in stool flora of rats,and reduce the abundance of Bacteroides in stool flora of rats,most of which are mainly beneficial bacteria.Simulated microgravity can increase the overall richness and diversity of mucosal flora,increase the abundance of Bacteroides and Desulphurides in the rat mucosal flora,and decrease the abundance of firmicutes,most of which are proteobacteria.After probiotics intervention,the overall Bacteroidetes trend in simulated microgravity rats was increased.CONCLUSION B.lactis BLa80 can ameliorate intestinal mucosal injury,regulate intestinal flora,inhibit ERK expression,and activate the SCF/c-kit signaling pathway,which may have a facilitating effect on gastrointestinal motility in simulated microgravity rats.展开更多
基金supported by the National Natural Science Foundation of China(81960851)Jiangxi Natural Science Foundation(20202BABL206132)Key Research Office of Traditional Chinese Medicine Syndrome Foundation of Jiangxi Administration of Traditional Chinese Medicine(8-4),and Science and Technology Innovation Team Development Program of Jiangxi University of Chinese Medicine(CXTD22016).
文摘Objective:To investigate the potential mechanism of Wendan decoction in obesity by screening target genes with promoter region methylation changes and constructing a multiple signaling pathways network based on promoter methylation.Methods:The methylation degree of Itgad,Col8a1,Adra2b,Jund,Rab2a,Wnt8b,Fzd9,B4galt7,Pik3cd,Creb1,Stard8,and Mmp1 in the abdominal adipose tissue of obese rats was determined using the Agena MassARRAY system.Western blot was performed to assess protein expression levels.Target genes were identified based on the methylation degree in the promoter region and protein expression.Enrichment analysis of signaling pathways was conducted to identify relevant target genes and obtain a multiple signaling pathway network associated with obesity.Core and terminal effector molecules in the pathway networks were selected as research targets for reverse transcription-polymerase chain reaction(RT-PCR)analysis.Results:Four genes(Adra2b,Creb1,Itgad,and Pik3cd)showed a degree of promoter methylation consistent with their respective protein expression levels.Among them,Adra2b,Creb1,and Pik3cd expression increased,while that of Itgad decreased.Enrichment analysis revealed that Creb1 and Pik3cd were involved in 6 signaling pathways related to obesity:tumor necrosis factor(TNF)signaling pathway,growth hormone synthesis/secretion and action,adenosine 5'-monophosphate-activated protein kinase(AMPK)signaling pathway,relaxin signaling pathway,cyclic nucleotide(cAMP)signaling pathway,and phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt)signaling pathway.Subsequently,a multiple signaling pathways network was constructed based on promoter methylation.Key molecules including protein kinase B(AKT),mechanistic target of rapamycin complex 1(mTORC1),and unc-51 like autophagy activating kinase 1(ULK1),as well as terminal effector molecules interleukin-1β(IL-1β),interleukin-6(IL-6),and chemokine(C-X-C motif)ligand 2(CXCL2)were selected as research targets.Wendan decoction decreased the expressions of AKT,mTORC1,IL-1β,IL-6,and CXCL2 while up-regulating ULK1 expression.Conclusion:The mechanism of Wendan decoction in preventing obesity involves the regulation of multiple signaling pathways through the control of Creb1 and Pik3cd gene promoter methylation.However,the associated multi-path gene regulation mechanism in preventing obesity is complex.Thus,further exploration is needed to elucidate the role of methylation changes in this mechanism.
基金supported by the National Natural Science Foundation of China(32471186,31771318)the 14^(th) Five-Year-Plan Advantageous and Characteristic Disciplines(Groups)of Colleges and Universities in Hubei Province for Exercise and Brain Science from Hubei Provincial Department of Education,and the Leading Talent Program Foundation from Wuhan Sports University to Ning Chen+3 种基金the National Natural Science Foundation of China(81701391)the Natural Science Foundation of Hubei Province(2023AFB700)Key Project of Scientific Research of Education Department of Hubei Province(D20234101)Young and Middle aged Scientific Research Team Project of Wuhan Sports University(21KT08)to Jingjing Fan.
文摘Obesity,caused by excessive energy,leads to body weight gain and various diseases,including cognitive impairment.Current studies suggest that diet restriction such as optimal fasting and regular exercise are crucial for improving cognitive capacity.However,further exploration is needed to understand the specific mechanisms of high fat diet(HFD)-induced cognitive decline in obesity.In the present study,4-month-old mice were subjected to HFD feeding for 18 weeks,followed by aerobic exercise and high-intensity intermittent exercise,regular diet feeding,and intermittent fasting for 8 weeks,and then used to evaluate cognitive capacity,inflammation,compromised insulin signaling pathway,and apoptosis in hippocampal tissue,as well as AMPK/SIRT1 and TLR4 signal pathways.Obese mice revealed impaired cognitive capacity as compared with mice fed with regular diets.In contrast,aerobic exercise,high-intensity intermittent exercise,regular diet,and intermittent fasting could inhibit apoptosis caused by inflammation-mediated compromised insulin signaling pathway in hippocampal tissues through activating the AMPK/SIRT1 signal pathway and suppressing the TLR4 signal pathway,thereby rescuing the cognitive impairment of obese mice.Therefore,diet restriction and exercise interventions may play a positive role in reverting obesity-induced cognitive impairment.
基金Supported by Naional Key Basic Research Program(973, No. 2009 CB 522905)
文摘Objective To observe influence of moxibustion at ＂Shenshu＂ （肾俞BL 23） and ＂Zusanli＂ （足三里 ST 36） on proteins expression of Ras, ERKI/2, and Raf in synovial tissues in rats with rheumatoid arthritis （RA） to elucidate molecular mechanisms of moxibustion on treating RA. Methods Thirty Wistar rats were randomly divided into a normal control group （Group Cont）, a model group （Group Mod） and a moxibustion group （Group Mox） with 10 rats in each group. The model was established by using both environmental factors such as wind, cold and damp and a biological factor （Freund＇s complete adjuvant）. Suspended moxibustion was applied at bilateral BL 23 and ST 36 for 20 min, applied once daily for 15 days, starting from the 3rd day after modeling. Volumes of metaleg digiti pedis on the right side were measured before, after and 15 days after modeling. Protein expressions of Ras, ERK 1/2, p-ERKI/2 and Raf in synovium tissues were detected by western blot. Results After modeling, volumes of metaleg digiti pedis on the right side were significantly enlarged, compared with those of rats in Group Cont （P〈0.01）. After treatment by moxibustion for 15 days, the metatarsale girth of metaleg digiti pedis on the right side in Group Mox was significantly reduced, compared with that in Group Mod （P〈0.01）. Protein expressions of Ras, Raf, ERK1/2 and p-ERK1/2 in Group Mod were significantly increased, compared with those in Group Cont （P〈0.01） and Group Mox （P〈0.01）. Protein expressions of Ras, Raf, ERK1/2 and p-ERK1/2 in Group Mox were significantly lowered, compared with those in Group Mod （all P〈0.01）. Conclusions Moxibustion can suppress excessive protein expressions of Ras, Raf and ERK1/2 in synovium tissues of rats with experimental RA to inhibit abnormal activation of Ras-MAPK signal Pathway, which may be one of the molecular mechanisms of moxibustion in alleviating synovitis.
文摘Objective To investigate effect of pinacidil, an ATP sensitive potassium channel （KATP） opener, on the neuronal apoptosis and its signaling transduction mechanism following focal cerebral ischemia-reperfusion in rats. Methods One hundred male Wistar rats were randomly divided into four groups： A, sham-operated group; B, ischemia-reperfusion group; C, KATe opener treatment group; and D, KATe opener and blocker treatment group. The middle cerebral artery occlusion （MCAO） model was established by using the intraluminal suture occlusion method, neuronal apoptosis was determined by TUNEL staining, and expressions of caspase-8, caspase-9 and caspase-3 mRNA were detected by in situ hybridization. Results （1） The numbers of apoptotic neurons at 12 h, 24 h, 48 h, and 72 h were significantly less in group C than in groups B and D （P 〈 0.01 or P 〈 0.05）; and there was no difference between groups B and D at all time points （P 〉 0.05）. （2） The expressions of caspase-3 mRNA and caspase-8 mRNA at all times and the expressions of caspase-9 mRNA at 12 h, 24 h, 48 h, 72 h were significantly lower in group C than in groups B and D （P 〈 0.01 or P 〈 0.05）; and there were no differences between groups B and D at all time points （P 〉 0.05）. Conclusions KATP opener can significantly decrease the neuronal apoptosis and the expressions of caspase-3, caspase-8 and caspase-9 mRNAs following cerebral ischemiareperfusion. The neuronal apoptosis may be decreased by the inhibition of both mitochondrial and death-receptor signal pathways.
基金supported by grants from the National Natural Science Foundation (31872979, 31572366)the National Key Research and Development Program of China (2017YFD0502002)the National Basic Research Programs of China (2015CB943102)。
文摘Background: Intramuscular fat(IMF) content is a vital parameter for assessing pork quality. Increasing evidence has shown that microRNAs(miRNAs) play an important role in regulating porcine IMF deposition. Here, a novel miRNA implicated in porcine IMF adipogenesis was found, and its effect and regulatory mechanism were further explored with respect to intramuscular preadipocyte proliferation and differentiation.Results: By porcine adipose tissue miRNA sequencing analysis, we found that miR-146a-5p is a potential regulator of porcine IMF adipogenesis. Further studies showed that miR-146a-5p mimics inhibited porcine intramuscular preadipocyte proliferation and differentiation, while the miR-146a-5p inhibitor promoted cell proliferation and adipogenic differentiation. Mechanistically, miR-146a-5p suppressed cell proliferation by directly targeting SMAD family member 4(SMAD4) to attenuate TGF-β signaling. Moreover, miR-146a-5p inhibited the differentiation of intramuscular preadipocytes by targeting TNF receptor-associated factor 6(TRAF6) to weaken the AKT/mTORC1 signaling downstream of the TRAF6 pathway.Conclusions: MiR-146a-5p targets SMAD4 and TRAF6 to inhibit porcine intramuscular adipogenesis by attenuating TGF-β and AKT/mTORC1 signaling, respectively. These findings provide a novel miRNA biomarker for regulating intramuscular adipogenesis to promote pork quality.
基金Supported by State Natural Science Fund (No. 30772685)
文摘OBJECTIVE： To observe the influence of Sijunzi decoction and Yupingfeng powder on the expression of the relevant DNAs of janus kinase （JAK）-signal transducer and activator of transcription （STAT） sig- nal pathway of the brain in spleen-deficiency mod- el rats. METHODS： Eighty male Wistar rats of sanitary degree were divided randomly into four groups： nor- mal group, model group, treatment group 1, treatment group 2. Besides the rats in the normal group, all the rats in other 3 groups were prepared as spleen deficiency model. The treatment group 1 were treated with Sijunzi decoction and the treat- ment group 2 were treated with Yupingfeng powder. After treatment for 6 weeks, perfusion was given and the brain was taken for detection of the ex- pression of the relevant DNAs of JAK-STAT signal pathwayofthe brain in SD rats bygene chip method. RESULTS： Spleen deficiency could lead to increase expression of JAKI, STATI and Interleukin 4 （IL-4） in the brain, but the decrease expression of Suppres- sor of cytokine signaling I （SOCSI）, prolactin recep- tor （PRLR） and binding protein 3 （GATA 3）. Sijunzi decoction could increase expression of STAT3, Pro- lactin （PRL） and GATA3, but decrease expression of JAKI, STAT, STAT4, Interleukin 10 receptor, alpha （ILIORA）, Coagulation factor II （F2）, PRLR, MAD homolog 3 （SMAD3） and IL-4. Yupingfeng powder could decrease expression of JAKI, STATI, STAT4, SOCS4_ predicted, Epidermal growth factor receptor （EGFR）, PRLR, High mobility group AT-hook I （HMGAI 0）, IL-4. CONCLUSION： Sijunzi decoction and Yupingfeng powder can improve immune function of the rat through influencing the genetic expression of JAK-STAT signal pathway.
基金Project supported by the Zhejiang Provincial Natural Science Foundation of China(No.LY15H150004)the Teaching Department of the Zhejiang Province(No.Y201330073),China
文摘Accumulating evidence indicates that endostatin inhibits fibrosis. However, the mechanism is yet to be clarified. The aim of this study is to evaluate the effect of endostatin on platelet-derived growth factor-BB （PDGF-BB）- or transforming growth factor β1 （TGF-β1）-induced fibrosis in cultured human skin fibroblasts, and to further examine the molecular mechanisms involved. Human dermal flbroblasts were cultured in Dulbecco＇s modified Eagle＇s medium （DMEM） and serum-starved for 48 h before treatment. Cells were grouped as follows： ＂PDGF-BB＂, ＂PDGF-BB＋ endostatin＂, ＂TGF-β1＂, ＂TGF-β1＋endostatin＂, ＂endostatin＂, and ＂blank control＂. The fibroblasts were stimulated with either TGF-β1 or PDGF-BB for 72 h in order to set up the fibrosis model in vitro. The cells were co-cultured with either TGF-β1 or PDGF-BB and endostatin and were used to check the inhibiting effect of endostatin. A blank control group and an endostatin group were used as negative control groups. The biomarkers of fibrosis, including the expression of collagen I, hydrroxyproline, and α-smooth muscle actin （a-SMA）, were evaluated using an enzyme-linked immune- sorbent assay （ELISA） and Western blot. The expression of phosphorylated PDGF receptor β （p-PDGFRβ）, PDGFRβ, phosphorylated extracellular signal-regulated kinase （p-ERK）, and ERK was detected using Western blot and im- munofiuorescent staining was used to explore the mechanisms. Both PDGF-BB and TGF-β1 significantly up-regulated the expression of collagen I, hydroxyproline, and a-SMA. Endostatin significantly attenuated both the PDGF-BB- and TGF-β1-induced over-expression of collagen I, hydroxyproline, and a-SMA. PDGF-BB and TGF-β1 both promoted the expression of PDGFR, ERK, and p-ERK. Endostatin inhibited the expression of PDGFR and p-ERK but did not affect the expression of total ERK. Endostatin inhibited hypertrophic scar by modulating the PDGFRI3/ERK pathway. En- dostatin could be a promising multi-target drug in future fibrosis therapy.
基金supported by National Natural Science Foundation of China(No.81560124)Hainan Key Research and Development Projects(ZDYF2018131,ZDYF2017113,ZDYF2017114)+1 种基金Hainan Science and Technology Planned Project of Youth Outstanding Ability of Innovation(201704)Hainan Health Family Planning Industry Project(13A210277)
文摘Objective:To investigate the role of oxidative stress in human renal tubular epithelial cells(HK-2)induced by high glucose and the underlying signal pathway in vitro.Methods:MYPT1,pro-caspase-3,PGC-1α,and Drpl protein expressions were measured by Western blot.MnSOD2,Drp1 and PGC-1αmRNA expressions were detected by real time PCR.Results:Results showed that high glucose significantly up-regulated the protein expressions of MYPT1,pro-caspase-3 and the mRNA expression of MnSOD2 in HK-2 cells;while Rho kinase inhibitor fasudil and ROCK1 siRNA inhibited protein expressions of pro-caspase-3 and the mRNA expression of MnSOD2 in HK-2 cells induced by high glucose.Importantly,fasudil and ROCK1 siRNA markedly inhibited the expressions of mitochondrial motor proteins Drp1 and mitochondrial gene PGC-la in HK-2 cell=s induced by high glucose.Conclusions:Our findings suggest that Rho kinase signal pathway is involved in mitochondrial oxidative damage and apoptosis in high glucose-induced renal tubular epithelial cells by regulating mitochondrial motor proteins Drp1 and mitochondrial gene PGC-1α.Targeting Rho kinase signal pathway might be a potential strategy for the treatment of diabetic nephropathy.
文摘The panax notoginseng saponin(PNS) had been clinically used for the treatment of cardiovascular diseases and stroke in China.It had been demonstrated that PNS could protect cardiomyocytes from injury induced by ischemi- a,but the underlying molecular mechanisms of this protective effect were still unclear.This study was aimed to investigate the protective effect and molecular mechanisms of PNS on apoptosis in H9c2 cells in vitro and rat myocardial ischemia injury model in vivo.Annexin-V/PI assay shew that PNS could protect H9c2 cells from apoptosis induced by serum, glucose and oxygen deprivation(SGOD) in a dose-dependent manner.However,the anti-apoptotic effect of PNS was reversed by LY294002,a specific PI3K inhibitor.This antiapoptotic effect of PNS was confirmed by JC-1,a specific probe of mitochondrial membrane potential staining.PNS could significantly increase phos-Akt in H9c2 cells by Western blot assays and its effect could be inhibited by LY294002.Furthermore,PNS could improve ischemic-induced left ventricular function as reflected by EF,LVDd and LVDs.PNS could also inhibited cellular apoptosis in myocardial tissues in ischemic rats by TUNEL assay.PNS administration also increased the expression of phos-Akt in rat ischemic myocardial tissues.These results suggested that PNS could protect myocardial cells from apoptosis induced by ischemia in vitro model and in vivo model through activating-PI3K/Akt signal pathway which may be meaningful for further understanding the molecular mechanisms of cardiac protection of PNS.And the results might be useful in treatment of myocardial ischemia in future.
基金Supported by the Provincial Innovation Team for Cataract and Ocular Fundus Disease in the Second People’s Hospital of Yunnan Province(No.2017HC010)the Key Laboratory of Yunnan Province for the Prevention and Treatment of Ophthalmology(No.2017DG008)Expert Workstation of Yao Ke(No.2017IC064)。
文摘AIM:To investigate the feasibility and mechanism of immune tolerance in allergic conjunctivitis.METHODS:The allergic conjunctivitis immune tolerance mice model was established by ragweed pollen(RW)and the related cytokines were detected.The mice were divided into 9 groups and the maslinic acid(MA)or PBS were given for different group after modeling.The expression levels of chemokine ligand 5(CCL5)and P-65 in the conjunctival tissue were analyzed by immunohistochemistry,quantitative reverse transcription polymerase chain reaction(q RT-PCR)and Western blot.The percentage of interleukin-17(IL-17)and CD4+CD25+in the splenocyte supernatant was analyzed by flow cytometry.Fur thermore,the serum and splenocyte supernatant concentration of total-IgE,interleukin-10(IL-10),and IL-17 was analyzed by enzyme linked immune response(ELISA).RESULTS:After the model was established,symptoms of conjunctivitis were alleviated,the level of P-65,CCL5,IL-17,and total-IgE was raised,while the expression of IL-10,CD4+CD25+was decreased.This result fully demonstrated that a typical IL-17/regulatory-T-cells(Treg cells)imbalance and NF-κB activation.When the NF-κB signal pathway was suppressed,it showed that there was a further relief of conjunctivitis in mice.At the same time,the expression of total-IgE,IL-17,and CCL5 was decreased and the expression of anti-inflammatory factor(IL-10,CD4+CD25+)was increased.CONCLUSION:In the state of immune tolerance,symptoms of conjunctivitis in mice are alleviated,the Th-17 cells of allergic conjunctivitis mice are inhibited,and Treg cells activity is enhanced.
基金the Research Project of National Natural Science Foundation of China(81960761,81960751,81680705)Basic Ability Improvement Project for Young and Middle-aged Teachers in Colleges and Universities of Guangxi(2018KY1149)Natural Science Foundation of Guangxi Province(2020JJA140257).
文摘Hepatic fibrosis is a reversible pathological phenomenon in the early and middle stages,but but no satisfactory intervention drugs have been available so far.Recent studies have suggested that microcirculation disturbance of liver is one of the important pathogenesis of chronic liver disease,the improvement of microcirculation is beneficial to the recovery of liver function and the delay of liver fibrosis.Hepatic stellate cells are the core cells of hepatic fibrosis,and also the most critical cells that affect the microcirculation of the liver.While TLR4/MyD88/NF-κB and TLR4/MyD88/MAPKs which are based on the action of hepatic stellate cells are two pathways that have very important influence on the inflammatory response of liver,the proliferation and apoptosis of hepatic stellate cells,and the secretion of fibrogenic cytokines.It was found that Plumbapin,the active ingredient of Guangxi specialty ethnic medicine,has the definite effect of promoting blood circulation and removing blood stasis and anti-hepatic fibrosis,but its mechanism is not clear.In this study,the research progress of the above problems was reviewed,and further research ideas were derived as follows:the pharmacological effect of Plumbapin on anti-hepatic fibrosis,promoting blood circulation and removing stasis was based on the influence of TLR4/MyD88/NF-κB and MAPKs signal pathway.
文摘[Objectives]To explore the effect and possible mechanism of bergenin in relieving allergic rhinitis(AR)in mice.[Methods]50 C57/BL6 mice were randomly divided into blank group(n=10),model group(n=10)and high(100 mg/kg),medium(50 mg/kg)and low(25 mg/kg)dose bergenin groups with 10 mice in each group.Except for the blank group,the other mice were sensitized by basic ways combined with attack to replicate the AR model.From the 15th d of modeling(from the second d after the end of the basic modeling),the drug group was given bergenin orally for 15 d,and the blank group and model group were given the same volume of normal saline once a day.24 h after the last establishment of the model,the content of interleukin 4(IL-4),IL-6,TNF-αand IL-1βin nasal lavage fluid and serum of mice in each group was detected by ELISA.The expression of TLR-4,NF-κB and p-NF-κB in nasal mucosa of mice was detected by Western blot.[Results]Compared with the blank group,the content of inflammatory factors IL-4,IL-6,TNF-αand IL-1βin nasal lavage fluid and serum of model group was significantly increased,and the protein expression of TLR-4 and p-NF-κB was significantly increased.After the intervention of bergenin,the content of IL-4,IL-6,TNF-αand IL-1βin nasal lavage fluid and serum and TLR-4 and p-NF-κB protein in tissue was significantly inhibited in bergenin group.[Conclusions]Bergenin can effectively reduce allergic inflammation in AR model mice,and its mechanism may be related to inhibition of inflammation and down-regulation of TLR-4/NF-κB signal pathway.
文摘Background To investigate the effects and mechanisms of cellular repressor of ElA stimulated genes (CREG) on endothelial cell(EC) migration.Methods vascular endothelial cells(VE),CREG overexpression VEs, CREG suppression VEs and VEs transfected with CREG gene modified adenovirus(Ad-CREG) were cultured with dulbecco’s modified eagle’s medium contained 10%fetal calf serum. Western blot was used to detect the protein level of CREG and integrin-linked kinase(ILK) in the four kind ECs.Tran-swell migration model was applied to compare the migration cell number of the four kind ECs.Two kinds of ILK mutant plasmids;PCXN2-flag-ILK wt-IRES-GFP(wild-type ILK)and PCXN2-flag-ILK p-parvin-IRES-GFP(P-parvin-binding mutant) were used to transfect VS and VE respectively,then the two kind transfection ECs were named as VS-wtILK and VE-P -parvin which were selected by G418(600ng/ml)for 2 weeks;Transwell migration model was applied to compare migration capability before and after ILK plasmids transfecting VE and VS.Results Western blot analysis showed that CREG overexpression promoted ILK expression in ECs,on the contrary,ILK expression was down-regulated in CREG silent ECs(P【0.05).Further more,ILK expression was up-regulated obviously in VE transfected with Ad-CREG(P【 0.05);Transwell migration model showed that EC’s migration capability was positively correlated with the expression level of CREG in EC,that is,CREG overexpression induced VE migration and CREG silent suppressed VE migration, moreover,Ad-CREG transfecting VE showed better migration capability accompanied with CREG expression increase by transwell migration model(P【0.05).In order to know the relationship between ILK expression and cell migration,we obtained stable transfection cell strains of VS-wtILK and VE-Pparvin, transwell migration model demonstrated that VS-wtILK remarkably corrected the poor migration capability of VS(P【 0.01),butβ-parvin combining site mutation in ILK genes inhibited VE migration markedly(P【0.01).Conclusions ILKp -parvin signal pathway mediated vascular endothelial cell migration induced by CREG.
基金Supported by the National Program on Key Basic Research Project(973 Program)(2012CB518900)the National Natural Science Foundation of China(31160240,31260621)+2 种基金the 12th Five Years Key Programs for Science and Technology Development of China(2012ZX10002006)the Hangzhou Normal University Supporting Project(PE13002004042)the Natural Science Foundation of China of Jiangxi(20114BAB204016)
文摘The melanoma differentiation-associated gene-7（mda-7）,IL-24,has the specific functions that induce cancer cell apoptosis without doing harm to normal cells. We systematically review the apoptotic signal pathways and their regulatory mechanisms induced by Ad.IL-24 and IL-24 in diverse cancer cells. IL-24 can participate in varied signal transduction pathways,including JAK,p38 MAPK,Wnt/β-catenin,JNK,ER stress and mitochondria-associated signal pathways. And we review five proteins interacting with IL-24,including Bip/GRP78,S1 R,PKR,Beclin1 and soluble clusterin,which are relative to the tumor-specific effect of IL-24. It is speculated that ER stress,G-protein pathways and MAPK signal pathways may be the primary upstream effectors which activate the sequential downstream mediators resulting in apoptosis induced by IL-24 in tumor cells. Experimental results also show that IL-24 sensitizes cancer cells and indirectly promotes apoptosis rather than functions as a direct apoptosis inducer itself.
文摘Psoriasis is a complex skin disease and the pathogenesis of psoriasis is not clear.The purpose of this study is to identify the key driving genes and signal pathways involved in psoriasis and to predict the potential miRNA,for further understanding the pathogenesis of psoriasis.Methods:Three gene expression profiling chips,including GSE67853,GSE78097,and GSE136757 with a total of 120 samples were collected and analyzed with R software.The protein-protein interaction network of differentially expressed genes was constructed with STRING database and Cytoscape.CIBERSORT was used to evaluate the infiltration of immune cells in psoriasis tissues,and the correlation between diagnostic markers and infiltrating immune cells was analyzed.Further,the key biomarkers were identified and the targeting miRNA of crucial genes was predicted.Results:A total of 201 differentially expressed genes(163 upregulated genes and 38 downregulated genes)were determined.CXCL1,CXCL2,and CXCL8,the critical biomarkers of psoriasis,were identified by different calculation methods.The potential critical signal pathway NOD-like receptor signaling pathway of psoriasis was explored by gene expression profiling chip gene enrichment analysis and differentially expressed gene enrichment analysis.Immune cell infiltration analysis found that CXCL1,CXCL2,CXCL8 was positively correlated with macrophages M1 and T cells CD4 memory activated and negatively correlated with macrophages M2 and mast cells resting.At the same time,through miRNA prediction,we found that hsa-miR-216a-3p and hsa-miR-6750-5p can be used as potential psoriasis targets.Conclusions:This research proposes a new comprehensive strategy to identify psoriasis’s potential biomarkers through cross-validation and significant scores of different calculation methods.In this research,we identified CXCL1,CXCL2,and CXCL8 as potential key biomarkers of psoriasis,and the NOD-like receptor signaling pathway is the critical signal pathway of psoriasis.Hsa-miR-216a-3p and hsa-miR-6750-5p can be used as potential psoriasis targets.
文摘Objective: to explore the effect of micro-current stimulation on activation or inhibition of relevant signal pathways (MAPK, Wnt/β-catenin, NF-κB) in LDH model rats. Methods: a total of 180 healthy adult SD rats, half male and half female, weighing 250-280g, were selected for modeling. After successful modeling, NPCs cells of intervertebral disc were divided into stimulation group and non-stimulation group according to the intervention of low-frequency electronic pulse current stimulation. The contents of MAPK, Wnt/β-catenin, NF-κB signal pathway and LDH-related characteristic indicators in the two groups of rats were compared. Results: the relative expressions of p-p38MAPK, p-ERK1, p-ERK2 and p-JNK in the stimulation group were significantly lower than those in the non-stimulation group (p 0.05). The mechanical stimulation pain threshold of the stimulation group was significantly higher than that of the non-stimulation group, and the content of IL-1β and TNF-α was significantly lower than that of the non-stimulation group (P 0.05). There was no significant difference in IL-6 between the two groups. Conclusion: microcurrent stimulation can reduce the relative expression of p-p38MAPK, p-ERK1, p-ERK2, p-JNK protein in rat dorsal root ganglion, suggesting that it can inhibit ERK signaling pathway in rats with lumbar disc herniation.
基金The project supported by National Natural Science Foundation of China(81573645,81603101,81473383)
文摘OBJECTIVE To investigate the protective effect of salvianolic acid A(Sal A)on isoproterenol-induced myocardial infarction in mice and its possible mechanisms.METHODS The mice were subcutaneously injected with isopropranol(ISO 8 mg·kg-1)to induce myocardial infarction and evaluated the myocardial protective effect of Sal A from mortality rate,electrocardiogram(ECG),heart function,myocardial infarction index,serum myocardial enzymes and explored its possible mechanisms from inflammatory,antioxidant and cells apoptosis.RESULTS Sal A can dose-dependently enhanced the heart function of myocardial infarction mice,reduced the heart index,inhibited the myocardial enzyme leakage,showed obvious myocardial protection effects.ELISA results showed that Sal A can reduce the expression of myocardial inflammatory cytokines such as IL-6,TNF-α.Western blotting confirmed that Sal A can increase the expression of anti-apoptotic proteins Bcl-2,reduce the expression of apoptosis protein Bax,and raise the phosphorylation level of PI3K and Akt.CONCLUSION Sal A have displayed significant protective effect against isoproterenol-induced myocardial infarction and its mechanism may be related to increasing of PI3K/Akt signal pathway and inhibition of cell apoptosis and inflammatory reaction.
基金National Natural Science Foundation of China(No.81673891,No.81560760)Guangxi Medical and Health Appropriate Technology Research and Development Project(No.S201532)Guangxi Zhuang Yao Pharmaceutical Preparation Improvement Project of Traditional Chinese Medicine(No.GZZJ16-03)。
文摘Heart failure(HF)is a kind of continuous development syndrome of cardiac insufficiency caused by various heart diseases.Not only does the prevalence continue to rise,but the mortality rate and readmission rate remain high.Heart failure is also the end-stage of cardiovascular disease and the main cause of death of patients,which seriously affects the health and quality of life of people all over the world.Ventricular remodeling plays a key role in the occurrence and development of heart failure.Therefore,by improving ventricular remodeling,it is of important research value to explore the intervention of traditional Chinese medicine in the development of heart failure.Studies have shown that the mediation of multiple signaling pathways can lead to progressive aggravation of ventricular remodeling,and experimental studies often confirm the therapeutic effects of traditional Chinese medicine.Traditional Chinese medicine usually achieves the therapeutic effect of heart failure through multiple targets and multiple approaches.In recent years,there have been more and more researches on the role and mechanism of Chinese medicine intervention in heart failure.However,it is concluded that Chinese medicine intervention has less influence on heart failure signal pathways.This article summarizes the understanding of Chinese medicine on heart failure and the five signal pathways related to Chinese medicine intervention in heart failure.The 5 signaling pathways in the world,namely transforming growth factor-β1(TGF-β1)/signal transduction protein(Smads)signaling pathway,Toll-like receptor(TLR)/nuclear transcription factor-κB(NF-κB)inflammation signaling pathway,Renin-angiotensinaldosterone(RAAS)system,phosphoinositide 3-kinase(PI3K)-serine/threonine protein kinase(AKT)signaling pathway and mitogen-activated protein kinase(MAPK)dependent signaling pathway.
文摘Objective: To observe the effect of acupuncture on the expression of ERM/PI3K/Akt signaling pathway in the brain tissue around the infarct focus of ischemic stroke model rats. Methods: According to the block randomization method, 80 male SD rats were divided into normal group (n = 10), sham operation group (n = 24), model group (n = 23) and acupuncture group (n = 23). The normal group was bred routinely without any intervention. In the sham operation group, only the skin was cut to find and the nerves and blood vessels were stripped and sutured. The model group and the acupuncture group were sutured to prepare the MCAO model. After the model was successfully established, acupuncture of the acupuncture group rats was given once a day, and the needles were kept for 30 minutes each time. The rats in the sham operation group and the model group only imitated capture and return, and were treated with acupuncture for 2 consecutive courses, each course was 5 days, and the two courses were rested for 2 days. After that, Western blot was used to detect ERM, changes in the concentration of PI3K and Akt proteins. Results: Compared with the normal group, the expression levels of ERM, PI3K, and Akt proteins in the sham operation group were reduced. Compared with the sham operation group, the expression levels of ERM, PI3K, and Akt proteins in the model group were significantly down-regulated. Compared with the model group, the protein expression levels of ERM, PI3K and Akt were significantly up-regulated in the acupuncture group, and the difference was statistically significant (P < 0.05). Conclusion: Acupuncture can significantly up-regulate the expression of ERM, PI3K and Akt proteins, and its effect mechanism may be related to the transduction of ERM/PI3K/Akt signaling pathway in cells.
文摘BACKGROUND Simulated microgravity environment can lead to gastrointestinal motility disturbance.The pathogenesis of gastrointestinal motility disorders is closely related to the stem cell factor(SCF)/c-kit signaling pathway associated with intestinal flora and Cajal stromal cells.Moreover,intestinal flora can also affect the regulation of SCF/c-kit signaling pathway,thus affecting the expression of Cajal stromal cells.Cajal cells are the pacemakers of gastrointestinal motility.AIM To investigate the effects of Bifidobacterium lactis(B.lactis)BLa80 on the intestinal flora of rats in simulated microgravity and on the gastrointestinal motility-related SCF/c-kit pathway.METHODS The internationally recognized tail suspension animal model was used to simulate the microgravity environment,and 30 rats were randomly divided into control group,tail suspension group and drug administration tail suspension group with 10 rats in each group for a total of 28 days.The tail group was given B.lactis BLa80 by intragastric administration,and the other two groups were given water intragastric administration,the concentration of intragastric administration was 0.1 g/mL,and each rat was 1 mL/day.Hematoxylin&eosin staining was used to observe the histopathological changes in each segment of the intestine of each group,and the expression levels of SCF,c-kit,extracellular signal-regulated kinase(ERK)and p-ERK in the gastric antrum of each group were detected by Western blotting and PCR.The fecal flora and mucosal flora of rats in each group were detected by 16S rRNA.RESULTS Simulated microgravity resulted in severe exfoliation of villi of duodenum,jejunum and ileum in rats,marked damage,increased space between villi,loose arrangement,shortened columnar epithelium of colon,less folds,narrower mucosal thickness,reduced goblet cell number and crypts,and significant improvement after probiotic intervention.Simulated microgravity reduced the expressions of SCF and c-kit,and increased the expressions of ERK and P-ERK in the gastric antrum of rats.However,after probiotic intervention,the expressions of SCF and ckit were increased,while the expressions of ERK and P-ERK were decreased,with statistical significance(P<0.05).In addition,simulated microgravity can reduce the operational taxonomic unit(OTU)of the overall intestinal flora of rats,B.lactis BLa80 can increase the OTU of rats,simulated microgravity can reduce the overall richness and diversity of stool flora of rats,increase the abundance of firmicutes in stool flora of rats,and reduce the abundance of Bacteroides in stool flora of rats,most of which are mainly beneficial bacteria.Simulated microgravity can increase the overall richness and diversity of mucosal flora,increase the abundance of Bacteroides and Desulphurides in the rat mucosal flora,and decrease the abundance of firmicutes,most of which are proteobacteria.After probiotics intervention,the overall Bacteroidetes trend in simulated microgravity rats was increased.CONCLUSION B.lactis BLa80 can ameliorate intestinal mucosal injury,regulate intestinal flora,inhibit ERK expression,and activate the SCF/c-kit signaling pathway,which may have a facilitating effect on gastrointestinal motility in simulated microgravity rats.
