Objectives:Ovarian cancer,a leading cause of gynecological malignancy-related mortality,is charac-terized by limited therapeutic options and a poor prognosis.Although pyrimethamine has emerged as a promising candidate...Objectives:Ovarian cancer,a leading cause of gynecological malignancy-related mortality,is charac-terized by limited therapeutic options and a poor prognosis.Although pyrimethamine has emerged as a promising candidate demonstrating efficacy in treating various tumors,the precise mechanisms of its antitumor effects remain obscure.This study was specifically designed to investigate the mode of action underlying the antitumor effects of pyrimethamine in preclinical settings.Methods:The effects of pyrimethamine on cellular proliferation were meticulously assessed using both the cell counting kit 8(CCK-8)assay and the colony formation assay,with the effects further confirmed in a murine model.A confocal microscope was utilized to monitor the dynamic alterations in mitochondria within ovarian cancer cells.Additionally,adenosine triphosphate(ATP)and reactive oxygen species(ROS)assays were conducted to measure mitochondrial damage induced by pyrimethamine in ovarian cancer cell lines.The mitochondrial membrane potential was assessed using fluorescent dyes as an indicator of mitochondrial functional status.Furthermore,transcriptome analysis and immunohistochemical techniques were employed to detect the impact of pyrimethamine on ovarian cancer cells.Results:Our results demonstrated that pyrimethamine induced ovarian cancer cell death through mitochondrial dysfunction and lethal mitophagy.Transcriptome profiling analysis and Western blot demonstrated that activation of the p38/JNK/ERK signaling pathway was implicated in the process of pyrimethamine-induced mitophagy in ovarian cancer cells.Importantly,combination treatment with pyrimethamine and paclitaxel in vitro and in vivo showed a synergistic antitumor effect.Conclusions:Altogether,these findings indicate that the antitumor effects of pyrimethamine result from the induction of lethal mitophagy via regulation of the p38/JNK/ERK pathway in ovarian cancer.Considering the low toxicity and high tolerance associated with pyrimethamine,it is suggested that pyrimethamine be evaluated in the treatment of ovarian cancer,either as a monotherapy or in combination with paclitaxel.展开更多
AIM： To identify the protein expression differences related to the CagA-induced ERK pathway activation in AGS cells. METHODS： Human AGS cells transfected with cagA and blank vector were treated with specific mitogen...AIM： To identify the protein expression differences related to the CagA-induced ERK pathway activation in AGS cells. METHODS： Human AGS cells transfected with cagA and blank vector were treated with specific mitogenactivated protein kinase kinase （MEK） inhibitor. Total cell proteins were combined by strong anion exchange （SAX2） and weak cation exchange （CM10） ProteinChip arrays and analyzed using surface-enhanced laser desorption/ ionization time-of-flight mass spectrometry （SELDI-TOF- MS） proteomics technology. Protein expression profiles were compared with those of inhibitor-untreated cagA transfectants. SwissProt/TrEMBL database searching for differentially expressed proteins was carried out using the TagIdent tool with the pI and mass information. RESULTS： When a total of 16 proteins that showed expression differences in inhibitor-untreated cagA transfectants were compared with vector transfectants, three proteins with m/z 4229, 8162 and 9084 were found to have no expression differences after treatment with MEK inhibitor, while the other 13 maintained the same expression differences after inhibitor treatment. Seven pieces of meaningful matching information for the three proteins were obtained from database searching. CONCLUSION： Biomarkers with m/z 4229, 8162 and 9084 are ERKI/2 phosphorylation dependent, andtherefore are the downstream molecules of ERK1/2 in the ERK/MAPK signaling pathway. The three biomarkers may be important cancer-associated proteins according to SwissProt/TrEMBL database information.展开更多
Dysregulated crosstalk between different signaling pathways contributes to tumor development,including resistance to cancer therapy.In the present study,we found that the mitogen-activated extracellular signal-regulat...Dysregulated crosstalk between different signaling pathways contributes to tumor development,including resistance to cancer therapy.In the present study,we found that the mitogen-activated extracellular signal-regulated kinase(MEK)inhibitor trametinib failed to suppress the proliferation of PANC-1 and MGC803 cells by activating the Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway,while the JAK2 inhibitor fedratinib failed to inhibit the growth of the PANC-1 cells upon stimulation of extracellular signal-regulated kinase(ERK)signaling.In particular,the most prominent enhancement of the anti-proliferative effect resulted from the concurrent blockage of the JAK2/STAT3 and ERK signaling pathways.Furthermore,the combination of the two inhibitors resulted in a reduced tumor burden in mice.Our evidence suggests novel crosstalk between JAK2/STAT3 and ERK signaling in gastric cancer(GC)and pancreatic ductal adenocarcinoma(PDAC)cells and provides a therapeutic strategy to overcome potential resistance in gastrointestinal cancer.展开更多
Background:Glioblastoma,a notably malignant tumor within the central nervous system,is distinguished by its aggressive behavior.Silvestrol,a robust inhibitor of the RNA helicase eukaryotic initiation factor 4A(eIF4A),...Background:Glioblastoma,a notably malignant tumor within the central nervous system,is distinguished by its aggressive behavior.Silvestrol,a robust inhibitor of the RNA helicase eukaryotic initiation factor 4A(eIF4A),has shown significant potential as an anticancer compound.Yet,the impact of silvestrol on glioblastoma,especially its molecular mechanisms,has not been fully elucidated.Methods:This investigation employed a variety of in vitro assays,such as cell counting kit-8(CCK-8),clonogenic,5-ethynyl-2′-deoxyuridine(EDU),wound healing,and flow cytometry,to evaluate cell cycle progression,apoptosis,cell viability,and migration.Western blot analysis was also performed to study the apoptosis and extracellular regulated kinase(ERK)pathways.After the ERK pathway was inhibited,differentially expressed genes(DEGs)in U87 cells were identified,followed by an analysis of target genes using the gene expression profiling interactive analysis(GEPIA)database.Results:Silvestrol significantly suppressed the proliferation,migration,and colony formation of glioma cells.It caused cell cycle arrest and enhanced apoptosis in these cells.Additionally,silvestrol stimulated the ERK pathway,with these effects being reversible by an ERK phosphorylation inhibitor.Transcriptome combined with GEPIA,GSCA,UALCAN,TIMER database screened 4 potential drug targets of silvestrol:chromosome 1 open reading frame 226(C1ORF226),mannosidase beta A(MANBA),IQ motif and Sec7 domain 2(IQSEC2),neuregulin 1(NRG-1).Among them,C1ORF226 was lower risk gene while MANBA,IQSEC2,and NRG-1 were high-risk genes.Furthermore,silvestrol notably reduced MANBA mRNA levels,which could be reversed by inhibiting ERK phosphorylation.Furthermore,silvestrol markedly decreased NRG-1 protein levels,with an additional reduction observed when the ERK pathway was blocked.Conclusion:Silvestrol’s anti-glioma effects are primarily due to the suppression of MANBA expression via the ERK pathway and possibly by hindering the translation of NRG-1 protein,thus reducing its expression.The downregulation of MANBA and NRG-1 proteins may be crucial in hindering glioma development and progression.These results highlight the intricate relationship between the ERK pathway and gene expression regulation in silvestrol’s therapeutic effectiveness against glioma.展开更多
Triple-negative breast cancer(TNBC)has a poor prognosis because of its aggressive behavior,absence of specific therapies,and high recurrence.1 The exact molecular mechanisms that drive the progression of TNBC are not ...Triple-negative breast cancer(TNBC)has a poor prognosis because of its aggressive behavior,absence of specific therapies,and high recurrence.1 The exact molecular mechanisms that drive the progression of TNBC are not yet fully understood,thus highlighting the urgent need for discovering novel potential treatment targets.展开更多
Background:Glioma is the most common tumor of the central nervous system with a poor prognosis.This study aims to explore the role of calcium/calmodulin-dependent protein kinase IIβ(CAMK2B)in regulating the malignant...Background:Glioma is the most common tumor of the central nervous system with a poor prognosis.This study aims to explore the role of calcium/calmodulin-dependent protein kinase IIβ(CAMK2B)in regulating the malignant progression of glioma cells,as well as the molecular mechanisms underlying these malignant behaviors.Methods:The correlation between CAMK2B expression in gliomas and patient prognosis was analyzed using immunohistochemistry,quantitative reverse transcription polymerase chain reaction(qRT-PCR),and western blot.Furthermore,the study explored the role of CAMK2B in glioma cell proliferation,invasion,and migration using cell counting kit-8(CCK-8),5-Ethynyl-2′-deoxyuridine(EdU),wound healing,transwell,and in vivo tumor xenograft assays.Result:Patients with high CAMK2B expression exhibited significantly better prognostic outcomes compared to those with low expression levels.Furthermore,CAMK2B expression was significantly lower in glioma tissues and cells compared to both normal brain tissue and human astrocyte cell lines.Notably,overexpression of CAMK2B in glioma cells led to an approximate 40%reduction in proliferative capacity and a 60–70%decrease in invasive and migratory abilities,compared to control glioma cells.These differences were statistically significant at p<0.05.Conversely,knockdown of CAMK2B using siRNA-CAMK2B significantly enhanced the proliferative,invasive,and migratory capabilities of glioma cells in both in vitro and in vivo settings,enhancing these abilities by 1.5 to 3 times.Notably,these effects were reversed through the application of the Rat Sarcoma viral oncogene homolog(Ras)pathway inhibitor,Salirasib.Western blot analysis revealed that knockdown of CAMK2B led to activation of the Ras/Rapidly Accelerated Fibrosarcoma(Raf)/Mitogen-activated protein kinase kinase(MEK)/Extracellular signal-regulated kinase(ERK)signaling pathway in glioma cell lines,whereas overexpression of CAMK2B resulted in the suppression of this pathway.Conclusion:CAMK2B inhibits glioma proliferation,invasion,andmigration through the Ras/Raf/MEK/ERK signaling pathway.展开更多
The published article titled“Procaine inhibits the proliferation and migration of colon cancer cells through inactivation of the ERK/MAPK/FAK pathways by regulation of RhoA”has been retracted from Oncology Research,...The published article titled“Procaine inhibits the proliferation and migration of colon cancer cells through inactivation of the ERK/MAPK/FAK pathways by regulation of RhoA”has been retracted from Oncology Research,Vol.26,No.2,2018,pp.209–217.展开更多
Objective:To study the regulatory effect of cryoablation on MAPK/ERK signaling pathway in mice with lung adenocarcinoma.Methods:Lewis mouse lung adenocarcinoma cell line was used to establish subcutaneous transplanted...Objective:To study the regulatory effect of cryoablation on MAPK/ERK signaling pathway in mice with lung adenocarcinoma.Methods:Lewis mouse lung adenocarcinoma cell line was used to establish subcutaneous transplanted tumor model in C57BL/6 mice.Ten mice were randomly divided into two groups:sham operation group and cryoablation group,with 5 mice in each group.The cryoablation group was treated with double circulation-rewarming ablation,and the sham operation group was treated with incision and suture at the transplanted tumor.The tumor tissues were taken 14 days after operation.Detect the effect of cryoablation on MAPK/ERK pathway related proteins by Western blot,such as KRAS,RAF1,MEK1,ERK1/2,P-RAF1,P-MEK1,P-ERK1/2.The expression of KRAS gene was further verified by qRt-PCR.Results:Compared with the sham operation group,the phosphorylated proteins P-RAF1,P-MEK1 and P-ERK1/2 in tumor tissue after cryoablation were decreased(P<0.05),and the key molecule KRAS in MAPK/ERK pathway was decreased in protein and gene expression(P<0.05).Conclusion:Cryoablation can negatively regulate MAPK/ERK signaling pathway by down-regulating KRas expression.展开更多
[Objectives]To explore the neuroprotective effects and mechanism of Longan Aril(LA)effective parts on PC12 cells injured by H2O2.[Methods]The neuroprotective effects of LA were evaluated by the cell viability,SOD and ...[Objectives]To explore the neuroprotective effects and mechanism of Longan Aril(LA)effective parts on PC12 cells injured by H2O2.[Methods]The neuroprotective effects of LA were evaluated by the cell viability,SOD and MDA content,apoptosis assay and relative protein expression of Aβand p-Tau.The neuroprotective mechanism of LA was studied by using metabolomics and network pharmacology,and the expressions of RAS/MEK/ERK signaling pathway-related proteins were detected by western blotting.[Results]LA could improve the cell survival rate and SOD content,and reduce apoptosis and expression of Aβand p-tau.Inhibition of RAS/MEK/ERK signaling pathway is a possible mechanism of LA neuroprotective effects.[Conclusions]LA has a neuroprotective effects in vitro and be likely to inhibit the process of AD by inhibition of RAS/MEK/ERK signalling pathway.展开更多
The differentiation of periodontal ligament(PDL)progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone.Vitamin C(VC),a water-soluble nutrient that cannot be biosynthesized by hum...The differentiation of periodontal ligament(PDL)progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone.Vitamin C(VC),a water-soluble nutrient that cannot be biosynthesized by humans,is vital for mesenchymal stem cells differentiation and plays an important role in bone remodeling.Therefore,the objective of this study was to determine the function and mechanism of VC in PDL progenitor cells osteogenic differentiation at the molecular level.We demonstrated that VC could induce the osteogenic differentiation and maturation of PDL progenitor cell without other osteogenic agents.During the process,VC preferentially activated ERK1/2 but did not affect JNK or p38.Co-treatment with ERK inhibitor effectively decreased the Vitamin C-induced expression of Runx2.ERK inhibitor also abrogated Vitamin C-induced the minimized nodules formation.PELP1,a nuclear receptor co-regulator,was up-regulated under VC treatment.PELP1 knockdown inhibited ERK phosphorylation.The overexpression of PELP1 had a positive relationship with Runx2 expression.Taken together,we could make a conclude that VC induces the osteogenic differentiation of PDL progenitor cells via PELP1-ERK axis.Our fi nding implies that VC may have a potential in the regeneration medicine and application to periodontitis treatment.展开更多
Objective： To investigate the relationship between extracellular signal-regulated kinase （ERK） pathway, multidrug resistance gene （mdr-1）, ribonucleotide recluctase M1 （RRM1） gene and their roles in gemcitabine...Objective： To investigate the relationship between extracellular signal-regulated kinase （ERK） pathway, multidrug resistance gene （mdr-1）, ribonucleotide recluctase M1 （RRM1） gene and their roles in gemcitabine （GEM） chemoresistance in pancreatic cancer cell line SW1990. Methods： The GEM-resistance cell model was constructed by a stepwise method. Immunohistochemistry was used to measure the expression of ERK protein （ERK1/2） in the established cell strains in a semiquantitative way. The mRNA expression of mdr-1 and RRM1 were detected by RT-PCR. MTT assay was performed to determine the IC50 value. Results： The established GEM-resistant cell strains were able to gain stable growth and passage ability in the medium contained different concentration levels of GEM （0, 30, 60, 100, 150 and 200 nmol/L）. The expression of ERK protein, mdr-1 and RRM1 gene were elevated accompanied by the increase of GEM concentration. There was a highly positive correlation between mdr-1, RRM1 expression and GEM-resistanca level （r = 0.960, P = 0.002 and r = 0.966, P = 0.002）. The expression of ERK protein also correlated with the mdr-1 and RRM1 level （r = -0.943, P = 0.005 and r = -0.883, P = 0.02）. At the GEM-resistance level of 200 nmol/L, the grey scale value of ERK1/2 was 164.22 ±13.17, mdr-1/β-actin and RRM1/β-actin were 1.41 ±0.04 and 1.45 ± 0.18, respectively; after treated with ERK pathway inhibitor U0126, these values synchronously decreased to 186.85 ± 13.14, 0.2 3± 0.02 and 0.21 ± 0.03, respectively; inversely, the ERK1/2 grey scale value was 106.55 ± 16.45, mdr-l/β-actin and RRMl/β-actin were 1.50± 0.07 and 1.52 ± 0.12, respectively, which presented a tendency of synchronously increase after treated with ERK pathway activator EGF. The IC50 values in GEM-resistant cells of 0 nmol/L and 200 nmol/L levels were 4.104 and 10.20, respectively. After treated with U0126, these values decreased to 3.26 and 4.50, respectively; while treated with EGF, the IC50 values increased to 8.89 and 17.17, respectively. Conclusion： The ERK pathway may induce the GEM-chemoresistance in pancreatic cell line SW1990 through the participation in the regulation of the mdr-1 and RRM1 gene expression.展开更多
BACKGROUND Insomnia is a disease where individuals cannot maintain a steady and stable sleep state or fail to fall asleep.Western medicine mainly uses sedatives and hypnotic drugs to treat insomnia,and long-term use i...BACKGROUND Insomnia is a disease where individuals cannot maintain a steady and stable sleep state or fail to fall asleep.Western medicine mainly uses sedatives and hypnotic drugs to treat insomnia,and long-term use is prone to drug resistance and other adverse reactions.Acupuncture has a good curative effect and unique advantages in the treatment of insomnia.AIM To explore the molecular mechanism of acupuncture at Back-Shu point for the treatment of insomnia.METHODS We first prepared a rat model of insomnia,and then carried out acupuncture for 7 consecutive days.After treatment,the sleep time and general behavior of the rats were determined.The Morris water maze test was used to assess the learning ability and spatial memory ability of the rats.The expression levels of inflammatory cytokines in serum and the hippocampus were detected by ELISA.qRTPCR was used to detect the mRNA expression changes in the ERK/NF-κB signaling pathway.Western blot and immunohistochemistry were carried out to evaluate the protein expression levels of RAF-1,MEK-2,ERK1/2 and NF-κB.RESULTS Acupuncture can prolong sleep duration,and improve mental state,activity,diet volume,learning ability and spatial memory.In addition,acupuncture increased the release of 1L-1β,1L-6 and TNF-αin serum and the hippocampus and inhibited the mRNA and protein expression of the ERK/NF-κB signaling pathway.CONCLUSION These findings suggest that acupuncture at Back-Shu point can inhibit the ERK/NF-κB signaling pathway and treat insomnia by increasing the release of inflammatory cytokines in the hippocampus.展开更多
This study aimed to evaluate the therapeutic properties of the traditional Chinese medicine Xuesanqi(XSQ,from the rhizome of Polygonum amplexicaule D.Don)in treating ulcerative colitis.We hypothesized that its many ac...This study aimed to evaluate the therapeutic properties of the traditional Chinese medicine Xuesanqi(XSQ,from the rhizome of Polygonum amplexicaule D.Don)in treating ulcerative colitis.We hypothesized that its many active components can alleviate symptoms of colitis by regulating the gut microbiota,its metabolites,and various signaling pathways.To test our hypotheses,we designed a DSS-induced colitis model in C57BL/6 male mice.Apparent metrics were evaluated in each group of mice and performed histological analysis of relevant tissues.The gut microbial composition was analyzed by 16S rRNA sequencing of bacteria.Simultaneously,the SCFAs content was detected by gas chromatography,inflammatory factor secretion was evaluated by ELISA or western-blot,the expression of tight junction protein and key proteins of the MAPK signaling pathway were analyzed by western-blot.Our result showed that the treatment with XSQ alleviated significant various symptoms such as weight loss,blood in stool,and shortening of colon.In addition,XSQ treatment restored the dysregulated gut microbiota in colitis mice,increased short chain fatty acids(SCFAs)and normalized the MAPK/ERK/JNK signaling pathways,promoted expression of tight junction protein Occludin,Claudin-1,and E-cadherin proteins.Furthermore,we also observed a dose-dependent pattern in these treatment responses.These findings demonstrated the active components of XSQ is a promising new treatment platform for ulcerative colitis.展开更多
Objectives In this work,we explore the effect of atorvastatin on myocardial apoptosis and caspase-8 acti-vation after coronary microembolization(CME)in rats.Methods Fifty rats were randomly divided into five groups;th...Objectives In this work,we explore the effect of atorvastatin on myocardial apoptosis and caspase-8 acti-vation after coronary microembolization(CME)in rats.Methods Fifty rats were randomly divided into five groups;the coronary microembolization(CME)group,the sham-operated(sham)control group,the gastric lavage control group,the atorvastatin lavage group,and the caspasse-8 inhibitor(N-acetyl-Ile-Glu-Thr-Asp-CHO,abbreviated as CHO)group,with 10 rats for each group.A microembolization ball was injected through the left ventricle for constructing the CME model.Animals in the sham control group were given an injection of physiological saline instead of the microembolization ball.Seven days before the operation,the atorvastatin group underwent gastric lavage with 20 mg/kg of atorvastatin once a day.Gastric lavage control animals underwent gastric lavage with an equivalent dose of physiological saline instead of the atorvastatin.Animals in the CHO group were given an intraperitoneal injection of 10 mg/kg of CHO 30 min before the operation.Six hours after the operation,cardiac ultrasonic detection was conducted on each group to measure the cardiac function indexes.TUNEL(Terminal-deoxynucleoitidyl transferase mediated dUTP nick end labeling)assays were used to measure myocardial apoptosis,and western blots were used to quantify the expression levels of activated caspase-3 and-8.Results(1)The echocardiographic parameters showed that,compared to the sham control animals,the left ventricular ejection fraction(LVEF)of the CME group was significantly decreased(P【0.05).In addition,cardiac sonography revealed a decrease in the left ventricular shortening fraction(FS)and cardiac output(CO),but an increase in the left ventricular end-diastolic dimension(LVEDd).Compared to the CME group,the atorvastatin and CHO groups exhibited significantly improved cardiac function(P【0.05).(2)When compared with the sham control,the myocardical apoptotic rate of the CME group,as well as the levels of activated caspase-3 and-8,increased significantly(P【0.05).The myocardial apoptotic rate,as well as the levels of activated caspase-3 and caspase-8 in the atorvastatin and CHO groups,decreased significantly(P【0.05)in comparison to the CME group.Conclusions The atorvastatin pretreatment clearly suppressed post-CME myocardial apoptosis and improved cardiac function.The most likely mechanism for these effects is the blockade of the myocardial death receptor-mediated apoptosis pathway.展开更多
Objective: To explore the therapeutic effect and underlying mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy (MN). Methods:Mice with MN was established by injecting cationic bovine serum album...Objective: To explore the therapeutic effect and underlying mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy (MN). Methods:Mice with MN was established by injecting cationic bovine serum albumin (c-BSA) into tail vein for several times. model mice were randomly divided into MN group (equal amount of distilled water), Shenqi Zhilong Decoction low dose group (12 g crude drug/kg), Shenqi Zhilong Decoction high dose group (24 g crude drug/kg), and Tripterygium wilfordii polyglycoside tablet group (14 mg/ kg). Another 10 un-treatment mice were taken as control group (equal amount of distilled water). The drug was administered orally once a day for 4 weeks. After the last administration, 24 hours urine was collected to determine the urinary protein content;blood from inner canthus was collected to measure the changes of kidney function, liver function, blood lipid and levels of IL-6, IL-4 and TNF-α in serum in each group;HE staining was used to observe the pathological changes of kidney. Immunohistochemical staining was used to observe the expression of IgG in kidney. The protein expression of ERK1/2 and cPLA2 in renal tissues was determined by Western-blot method. The gene expression of Neph1, Nephrin and Podocin mRNA in kidney tissues were detected by RT-PCR. Results: Compared with model group, Shenqi Zhilong decoction at low-dose and high-dose could significantly reduce the value of urine protein in MN mice;Decreased TC and TG levels (P<0.05 or P<0.01);Increased the levels of ALB and TP in liver function (P<0.05 or P<0.01);has no significant effects on the levels of CRE, UREA and UA in renal function (P>0.05). Decreased the contents of IL-6, IL-4 and TNF-α in serum (P<0.05 or P<0.01);Significantly down-regulated the protein expression levels of p-ERK1/2 and p-cPLA2 in kidney tissues of MN mice (P<0.05 or P<0.01);Significantly increased the expression levels of NephP1, Nephrin and Podocin mRNA in renal tissues (P<0.01). Conclusion: Shenqi Zhulong Decoction has a good therapeutic effect on MN mice, and the mechanism of action is related to regulate the expression of related genes of Nephrin-Podocin-Neph1 receptor complex for protecting the glomerular filtration barrier, and inhibite the activation of ERK/cPLA2 pathway for relieving damage of GEC and reduceing secretion of pro-inflammatory cytokines.展开更多
Cardiac fibrosis is a cause of morbidity and mortality in people with heart disease.Anti-fibrosis treatment is a significant therapy for heart disease,but there is still no thorough understanding of fibrotic mechanism...Cardiac fibrosis is a cause of morbidity and mortality in people with heart disease.Anti-fibrosis treatment is a significant therapy for heart disease,but there is still no thorough understanding of fibrotic mechanisms.This study was carried out to ascertain the functions of cytokine receptor-like factor 1(CRLF1)in cardiac fibrosis and clarify its regulatory mechanisms.We found that CRLF1 was expressed predominantly in cardiac fibroblasts.Its expression was up-regulated not only in a mouse heart fibrotic model induced by myocardial infarction,but also in mouse and human cardiac fibroblasts provoked by transforming growth factor-β1(TGF-β1).Gain-and loss-of-function experiments of CRLF1 were carried out in neonatal mice cardiac fibroblasts(NMCFs)with or without TGF-β1 stimulation.CRLF1 overexpression increased cell viability,collagen production,cell proliferation capacity,and myofibroblast transformation of NMCFs with or without TGF-β1 stimulation,while silencing of CRLF1 had the opposite effects.An inhibitor of the extracellular signal-regulated kinase 1/2(ERK1/2)signaling pathway and different inhibitors of TGF-β1 signaling cascades,comprising mothers against decapentaplegic homolog(SMAD)-dependent and SMAD-independent pathways,were applied to investigate the mechanisms involved.CRLF1 exerted its functions by activating the ERK1/2 signaling pathway.Furthermore,the SMAD-dependent pathway,not the SMAD-independent pathway,was responsible for CRLF1 up-regulation in NMCFs treated with TGF-β1.In summary,activation of the TGF-β1/SMAD signaling pathway in cardiac fibrosis increased CRLF1 expression.CRLF1 then aggravated cardiac fibrosis by activating the ERK1/2 signaling pathway.CRLF1 could become a novel potential target for intervention and remedy of cardiac fibrosis.展开更多
Objective:To observe the effect of Yishen Sanjie Huayu compound prescription on ERK/NF-κB signaling pathway in IgA nephropathy(IgAN)rats,and explore its effect on preventing and treating IgA nephropathy intrarenal ar...Objective:To observe the effect of Yishen Sanjie Huayu compound prescription on ERK/NF-κB signaling pathway in IgA nephropathy(IgAN)rats,and explore its effect on preventing and treating IgA nephropathy intrarenal arteriole disease.Methods:Fifty-five male SD rats were randomly divided into blank group,model group,ShenfukangⅡcapsule group and Losartan potassium tablet group.Bovine serum albumin(BSA)was used for intragastric administration and carbon tetrachloride(CCl4).IgAN rat model was established by subcutaneous injection and lipopolysaccharide(LPS)tail vein injection.ShenfukangⅡcapsule group and Losartan potassium tablet group were given each drug suspension 2ml/head/d one week after modeling Gavage was started.The blank group and the model group were given an equal volume of normal saline.The 24h urine protein(UTP)of the rats was measured at 4,8,and 12 weeks after the administration,and the blood creatinine(SCr)was measured after 12 weeks.,urea nitrogen(BUN),aldosterone(ADS),angiotensinⅡ(AngⅡ),immunohistochemical Envi-sion System two-step method to detect vascular endothelial growth factor(VEGF)and human matrix in the whole rat kidney and small artery area The expression of metalloproteinase-9(MMP-9),proliferating cell nuclear antigen(PCNA),extracellular regulatory protein kinase(ERK)1/2,nuclear transcription factor-κB(NF-κB),and the small arteries of rat kidney tissue The intima,media,vessel wall/vascular outer diameter value.Results:Compared with the model group,the expressions of VEGF,MMP-9,PCNA,ERK1/2 and NF-κB in kidney tissues of the ShenfukangⅡcapsule group and the Losartan potassium tablet group decreased(P<0.05),24hUTP and SCr,BUN level decreased(P<0.05),kidney tissue damage was alleviated;intima and vessel wall/vascular outer diameter values were significantly reduced(P<0.01),there was no significant difference in ADS between the groups.The AngⅡof the Tanpotassium tablets group was lower than that of the model group(P<0.05).Conclusion:Yishen Sanjie Huayu compound can inhibit the ERK/NF-κB signaling pathway in rats with IgA nephropathy,reduce the levels of VEGF,MMP-9,PCNA,ERK1/2,NF-κB,and inhibit intrarenal arteriole vascular endothelial cells Proliferate and reduce kidney damage.展开更多
Coreopsis tinctoria Nutt.,an edible flowering plant,belongs to the Chrysanthemum family and is mainly grown at high altitudes in Northwestern China.It is rich in polyphenolic compounds,particularly marein and flavomar...Coreopsis tinctoria Nutt.,an edible flowering plant,belongs to the Chrysanthemum family and is mainly grown at high altitudes in Northwestern China.It is rich in polyphenolic compounds,particularly marein and flavomarein,and possesses multiple health-promoting properties,such as antioxidant,hypoglycemic and vasorelaxant effects.Previous bioactivity investigations majorly focused on C.tinctoria and its crude extract.The aim of the present study was to prepare marein-dominant C.tinctoria flavonoids(CF),further investigate the CF protective effects of liver fibrosis induced by carbon tetrachloride and elucidate the associated molecular mechanisms.Results have demonstrated that CF effectively attenuated hepatofibrogenesis by increasing the activity of glutathione(GSH)and superoxide dismutase(SOD);suppressing the hepatic stellate cell(HSC)activation,inhibiting transforming growth factorβ(TGF-β)activation and the production ofα-smooth muscle actin(α-SMA),alleviating the phosphorylation of extracellular signal-regulated kinases1/2(ERK1/2)and small mothers against decapentaplegic1/2(Smad1/2),thus maintaining the collagen metabolic homeostasis in the liver.Our study revealed that CF possesses an efficacious protective effect against chronic hepatic fibrosis due to their strong inhibitory effects of oxidative stress and chronic inflammation.展开更多
Hypertension represents a significant chronic non-infectious disease in China,where Qingda Granule(QDG)has traditionally been employed for its management.However,the mechanisms underlying QDG's kidney protective e...Hypertension represents a significant chronic non-infectious disease in China,where Qingda Granule(QDG)has traditionally been employed for its management.However,the mechanisms underlying QDG's kidney protective effects remain incompletely understood.This study investigates QDG's role in ameliorating hypertensive kidney injury(KI)and elucidates the associated mechanisms.Network analysis identified potential therapeutic targets related to mitochondrial function and the extracellular signal-regulated kinase(ERK)cascade.Ribonucleic acid(RNA)sequencing revealed differentially expressed genes(DEGs)in hypertensive mouse kidneys,which were enriched in mitochondrial-related functions and normalized by QDG treatment.QDG attenuated angiotensin II(Ang II)-induced blood pressure elevation and enhanced renal artery flow.Both cellular and animal experiments demonstrated that QDG inhibits the ERK/ribosomal S6 kinase 1(RSK1)signaling axis,thereby preventing Ang II-induced mitochondrial damage and renal cell apoptosis.ERK pathway inhibitors confirmed QDG's mechanism of action through the ERK/RSK1 pathway.These findings indicate that QDG ameliorates hypertensive KI by preserving mitochondrial function through modulation of the ERK/RSK1 network,presenting a novel therapeutic approach for managing hypertensive KI in clinical practice.展开更多
The decline in oocyte quality and developmental potential with female reproductive aging is well recognized,yet the underlying mechanisms remain insufficiently investigated.In this study,an integrative analysis of tra...The decline in oocyte quality and developmental potential with female reproductive aging is well recognized,yet the underlying mechanisms remain insufficiently investigated.In this study,an integrative analysis of transcriptomes and morphologies of individual oocytes from young and aged mice identifies morphologically defective aged oocytes with distinct transcriptomic features.Further analysis demonstrates that both apoptotic and ferroptotic pathways are activated in the defective aged oocytes,and simultaneously blocking both pathways reverses the defective morphology to the largest extent.The Plat gene,which encodes tissue-type plasminogen activator(t PA),is downregulated with oocyte aging,and Plat knockdown increases oocyte susceptibility to both apoptosis and ferroptosis.Mechanistically,t PA functions as an upstream signaling molecule for Erk1/2 activation by interacting with particular phosphorylation kinases such as Alk.Consequently,Plat loss downregulates Erk1/2 pathway activity in oocytes,leading to degeneration through PCD.Supplementing exogenous t PA in vitro oocyte maturation cultures reduces the defect rate of aged oocytes,thereby improving oocyte quality and developmental potential.Collectively,Plat plays a pivotal role in protecting aged mouse oocytes from PCD,and t PA supplementation may serve as a potential clinical strategy to enhance oocyte quality in females of advanced maternal age.展开更多
基金supported by the Natural Science Foundation of Sichuan Province,China,grant number:2021YJ0011.
文摘Objectives:Ovarian cancer,a leading cause of gynecological malignancy-related mortality,is charac-terized by limited therapeutic options and a poor prognosis.Although pyrimethamine has emerged as a promising candidate demonstrating efficacy in treating various tumors,the precise mechanisms of its antitumor effects remain obscure.This study was specifically designed to investigate the mode of action underlying the antitumor effects of pyrimethamine in preclinical settings.Methods:The effects of pyrimethamine on cellular proliferation were meticulously assessed using both the cell counting kit 8(CCK-8)assay and the colony formation assay,with the effects further confirmed in a murine model.A confocal microscope was utilized to monitor the dynamic alterations in mitochondria within ovarian cancer cells.Additionally,adenosine triphosphate(ATP)and reactive oxygen species(ROS)assays were conducted to measure mitochondrial damage induced by pyrimethamine in ovarian cancer cell lines.The mitochondrial membrane potential was assessed using fluorescent dyes as an indicator of mitochondrial functional status.Furthermore,transcriptome analysis and immunohistochemical techniques were employed to detect the impact of pyrimethamine on ovarian cancer cells.Results:Our results demonstrated that pyrimethamine induced ovarian cancer cell death through mitochondrial dysfunction and lethal mitophagy.Transcriptome profiling analysis and Western blot demonstrated that activation of the p38/JNK/ERK signaling pathway was implicated in the process of pyrimethamine-induced mitophagy in ovarian cancer cells.Importantly,combination treatment with pyrimethamine and paclitaxel in vitro and in vivo showed a synergistic antitumor effect.Conclusions:Altogether,these findings indicate that the antitumor effects of pyrimethamine result from the induction of lethal mitophagy via regulation of the p38/JNK/ERK pathway in ovarian cancer.Considering the low toxicity and high tolerance associated with pyrimethamine,it is suggested that pyrimethamine be evaluated in the treatment of ovarian cancer,either as a monotherapy or in combination with paclitaxel.
基金National High Technology 2006AA02Z341 & NSFC 30430730
文摘AIM： To identify the protein expression differences related to the CagA-induced ERK pathway activation in AGS cells. METHODS： Human AGS cells transfected with cagA and blank vector were treated with specific mitogenactivated protein kinase kinase （MEK） inhibitor. Total cell proteins were combined by strong anion exchange （SAX2） and weak cation exchange （CM10） ProteinChip arrays and analyzed using surface-enhanced laser desorption/ ionization time-of-flight mass spectrometry （SELDI-TOF- MS） proteomics technology. Protein expression profiles were compared with those of inhibitor-untreated cagA transfectants. SwissProt/TrEMBL database searching for differentially expressed proteins was carried out using the TagIdent tool with the pI and mass information. RESULTS： When a total of 16 proteins that showed expression differences in inhibitor-untreated cagA transfectants were compared with vector transfectants, three proteins with m/z 4229, 8162 and 9084 were found to have no expression differences after treatment with MEK inhibitor, while the other 13 maintained the same expression differences after inhibitor treatment. Seven pieces of meaningful matching information for the three proteins were obtained from database searching. CONCLUSION： Biomarkers with m/z 4229, 8162 and 9084 are ERKI/2 phosphorylation dependent, andtherefore are the downstream molecules of ERK1/2 in the ERK/MAPK signaling pathway. The three biomarkers may be important cancer-associated proteins according to SwissProt/TrEMBL database information.
基金the Zhejiang Provincial Natural Science Foundation of China(Nos.LQ18H280005 and LY21H030002)the National Natural Science Foundation of China(Nos.81770535,81600595,81503297,81603340,81773945,and 81803775)+1 种基金the Medical Health Science and Technology Project of Zhejiang Provincial Health Commission(Nos.2019RC228,2019RC229,and 2019RC113)the Open Foundation from Chinese and Western Integrative Medicine in the Most Important Subjects of Zhejiang(No.ZXYJH2018002),China。
文摘Dysregulated crosstalk between different signaling pathways contributes to tumor development,including resistance to cancer therapy.In the present study,we found that the mitogen-activated extracellular signal-regulated kinase(MEK)inhibitor trametinib failed to suppress the proliferation of PANC-1 and MGC803 cells by activating the Janus kinase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway,while the JAK2 inhibitor fedratinib failed to inhibit the growth of the PANC-1 cells upon stimulation of extracellular signal-regulated kinase(ERK)signaling.In particular,the most prominent enhancement of the anti-proliferative effect resulted from the concurrent blockage of the JAK2/STAT3 and ERK signaling pathways.Furthermore,the combination of the two inhibitors resulted in a reduced tumor burden in mice.Our evidence suggests novel crosstalk between JAK2/STAT3 and ERK signaling in gastric cancer(GC)and pancreatic ductal adenocarcinoma(PDAC)cells and provides a therapeutic strategy to overcome potential resistance in gastrointestinal cancer.
基金This research was supported by the Chongqing Science and Health Joint Medical Research Project(2020FYYX150).
文摘Background:Glioblastoma,a notably malignant tumor within the central nervous system,is distinguished by its aggressive behavior.Silvestrol,a robust inhibitor of the RNA helicase eukaryotic initiation factor 4A(eIF4A),has shown significant potential as an anticancer compound.Yet,the impact of silvestrol on glioblastoma,especially its molecular mechanisms,has not been fully elucidated.Methods:This investigation employed a variety of in vitro assays,such as cell counting kit-8(CCK-8),clonogenic,5-ethynyl-2′-deoxyuridine(EDU),wound healing,and flow cytometry,to evaluate cell cycle progression,apoptosis,cell viability,and migration.Western blot analysis was also performed to study the apoptosis and extracellular regulated kinase(ERK)pathways.After the ERK pathway was inhibited,differentially expressed genes(DEGs)in U87 cells were identified,followed by an analysis of target genes using the gene expression profiling interactive analysis(GEPIA)database.Results:Silvestrol significantly suppressed the proliferation,migration,and colony formation of glioma cells.It caused cell cycle arrest and enhanced apoptosis in these cells.Additionally,silvestrol stimulated the ERK pathway,with these effects being reversible by an ERK phosphorylation inhibitor.Transcriptome combined with GEPIA,GSCA,UALCAN,TIMER database screened 4 potential drug targets of silvestrol:chromosome 1 open reading frame 226(C1ORF226),mannosidase beta A(MANBA),IQ motif and Sec7 domain 2(IQSEC2),neuregulin 1(NRG-1).Among them,C1ORF226 was lower risk gene while MANBA,IQSEC2,and NRG-1 were high-risk genes.Furthermore,silvestrol notably reduced MANBA mRNA levels,which could be reversed by inhibiting ERK phosphorylation.Furthermore,silvestrol markedly decreased NRG-1 protein levels,with an additional reduction observed when the ERK pathway was blocked.Conclusion:Silvestrol’s anti-glioma effects are primarily due to the suppression of MANBA expression via the ERK pathway and possibly by hindering the translation of NRG-1 protein,thus reducing its expression.The downregulation of MANBA and NRG-1 proteins may be crucial in hindering glioma development and progression.These results highlight the intricate relationship between the ERK pathway and gene expression regulation in silvestrol’s therapeutic effectiveness against glioma.
基金supported by the National Natural Science Foundation of China(No.82072923,82102829)Zhejiang Provincial Medical Science and Technology Program(China)(No.2020KY903).
文摘Triple-negative breast cancer(TNBC)has a poor prognosis because of its aggressive behavior,absence of specific therapies,and high recurrence.1 The exact molecular mechanisms that drive the progression of TNBC are not yet fully understood,thus highlighting the urgent need for discovering novel potential treatment targets.
基金supported by the Natural Science Foundation of Hebei Province(H2021206037)the Government-funded Project on Training of Outstanding Clinical Medical Personnel of Hebei Province in the year 2021(303-16-20-06)the Medical Research Project of Hebei Provincial Health Commission(20230031).
文摘Background:Glioma is the most common tumor of the central nervous system with a poor prognosis.This study aims to explore the role of calcium/calmodulin-dependent protein kinase IIβ(CAMK2B)in regulating the malignant progression of glioma cells,as well as the molecular mechanisms underlying these malignant behaviors.Methods:The correlation between CAMK2B expression in gliomas and patient prognosis was analyzed using immunohistochemistry,quantitative reverse transcription polymerase chain reaction(qRT-PCR),and western blot.Furthermore,the study explored the role of CAMK2B in glioma cell proliferation,invasion,and migration using cell counting kit-8(CCK-8),5-Ethynyl-2′-deoxyuridine(EdU),wound healing,transwell,and in vivo tumor xenograft assays.Result:Patients with high CAMK2B expression exhibited significantly better prognostic outcomes compared to those with low expression levels.Furthermore,CAMK2B expression was significantly lower in glioma tissues and cells compared to both normal brain tissue and human astrocyte cell lines.Notably,overexpression of CAMK2B in glioma cells led to an approximate 40%reduction in proliferative capacity and a 60–70%decrease in invasive and migratory abilities,compared to control glioma cells.These differences were statistically significant at p<0.05.Conversely,knockdown of CAMK2B using siRNA-CAMK2B significantly enhanced the proliferative,invasive,and migratory capabilities of glioma cells in both in vitro and in vivo settings,enhancing these abilities by 1.5 to 3 times.Notably,these effects were reversed through the application of the Rat Sarcoma viral oncogene homolog(Ras)pathway inhibitor,Salirasib.Western blot analysis revealed that knockdown of CAMK2B led to activation of the Ras/Rapidly Accelerated Fibrosarcoma(Raf)/Mitogen-activated protein kinase kinase(MEK)/Extracellular signal-regulated kinase(ERK)signaling pathway in glioma cell lines,whereas overexpression of CAMK2B resulted in the suppression of this pathway.Conclusion:CAMK2B inhibits glioma proliferation,invasion,andmigration through the Ras/Raf/MEK/ERK signaling pathway.
文摘The published article titled“Procaine inhibits the proliferation and migration of colon cancer cells through inactivation of the ERK/MAPK/FAK pathways by regulation of RhoA”has been retracted from Oncology Research,Vol.26,No.2,2018,pp.209–217.
基金The Fundamental Research Funds for the Central Universities(No.2020-JYB-ZDGG-127)National Key R&D Program of China(No.2018YFC1705102)。
文摘Objective:To study the regulatory effect of cryoablation on MAPK/ERK signaling pathway in mice with lung adenocarcinoma.Methods:Lewis mouse lung adenocarcinoma cell line was used to establish subcutaneous transplanted tumor model in C57BL/6 mice.Ten mice were randomly divided into two groups:sham operation group and cryoablation group,with 5 mice in each group.The cryoablation group was treated with double circulation-rewarming ablation,and the sham operation group was treated with incision and suture at the transplanted tumor.The tumor tissues were taken 14 days after operation.Detect the effect of cryoablation on MAPK/ERK pathway related proteins by Western blot,such as KRAS,RAF1,MEK1,ERK1/2,P-RAF1,P-MEK1,P-ERK1/2.The expression of KRAS gene was further verified by qRt-PCR.Results:Compared with the sham operation group,the phosphorylated proteins P-RAF1,P-MEK1 and P-ERK1/2 in tumor tissue after cryoablation were decreased(P<0.05),and the key molecule KRAS in MAPK/ERK pathway was decreased in protein and gene expression(P<0.05).Conclusion:Cryoablation can negatively regulate MAPK/ERK signaling pathway by down-regulating KRas expression.
基金Liaoning Natural Science Foundation(20180530033)Open Fund of Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications,Liaoning University of Traditional Chinese Medicine。
文摘[Objectives]To explore the neuroprotective effects and mechanism of Longan Aril(LA)effective parts on PC12 cells injured by H2O2.[Methods]The neuroprotective effects of LA were evaluated by the cell viability,SOD and MDA content,apoptosis assay and relative protein expression of Aβand p-Tau.The neuroprotective mechanism of LA was studied by using metabolomics and network pharmacology,and the expressions of RAS/MEK/ERK signaling pathway-related proteins were detected by western blotting.[Results]LA could improve the cell survival rate and SOD content,and reduce apoptosis and expression of Aβand p-tau.Inhibition of RAS/MEK/ERK signaling pathway is a possible mechanism of LA neuroprotective effects.[Conclusions]LA has a neuroprotective effects in vitro and be likely to inhibit the process of AD by inhibition of RAS/MEK/ERK signalling pathway.
基金This work is supported by grants:the National Basic Research Program(973 Program)(No.2011CB707705)National Natural Science Foundation of China(Grant No.11202229).
文摘The differentiation of periodontal ligament(PDL)progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone.Vitamin C(VC),a water-soluble nutrient that cannot be biosynthesized by humans,is vital for mesenchymal stem cells differentiation and plays an important role in bone remodeling.Therefore,the objective of this study was to determine the function and mechanism of VC in PDL progenitor cells osteogenic differentiation at the molecular level.We demonstrated that VC could induce the osteogenic differentiation and maturation of PDL progenitor cell without other osteogenic agents.During the process,VC preferentially activated ERK1/2 but did not affect JNK or p38.Co-treatment with ERK inhibitor effectively decreased the Vitamin C-induced expression of Runx2.ERK inhibitor also abrogated Vitamin C-induced the minimized nodules formation.PELP1,a nuclear receptor co-regulator,was up-regulated under VC treatment.PELP1 knockdown inhibited ERK phosphorylation.The overexpression of PELP1 had a positive relationship with Runx2 expression.Taken together,we could make a conclude that VC induces the osteogenic differentiation of PDL progenitor cells via PELP1-ERK axis.Our fi nding implies that VC may have a potential in the regeneration medicine and application to periodontitis treatment.
文摘Objective： To investigate the relationship between extracellular signal-regulated kinase （ERK） pathway, multidrug resistance gene （mdr-1）, ribonucleotide recluctase M1 （RRM1） gene and their roles in gemcitabine （GEM） chemoresistance in pancreatic cancer cell line SW1990. Methods： The GEM-resistance cell model was constructed by a stepwise method. Immunohistochemistry was used to measure the expression of ERK protein （ERK1/2） in the established cell strains in a semiquantitative way. The mRNA expression of mdr-1 and RRM1 were detected by RT-PCR. MTT assay was performed to determine the IC50 value. Results： The established GEM-resistant cell strains were able to gain stable growth and passage ability in the medium contained different concentration levels of GEM （0, 30, 60, 100, 150 and 200 nmol/L）. The expression of ERK protein, mdr-1 and RRM1 gene were elevated accompanied by the increase of GEM concentration. There was a highly positive correlation between mdr-1, RRM1 expression and GEM-resistanca level （r = 0.960, P = 0.002 and r = 0.966, P = 0.002）. The expression of ERK protein also correlated with the mdr-1 and RRM1 level （r = -0.943, P = 0.005 and r = -0.883, P = 0.02）. At the GEM-resistance level of 200 nmol/L, the grey scale value of ERK1/2 was 164.22 ±13.17, mdr-1/β-actin and RRM1/β-actin were 1.41 ±0.04 and 1.45 ± 0.18, respectively; after treated with ERK pathway inhibitor U0126, these values synchronously decreased to 186.85 ± 13.14, 0.2 3± 0.02 and 0.21 ± 0.03, respectively; inversely, the ERK1/2 grey scale value was 106.55 ± 16.45, mdr-l/β-actin and RRMl/β-actin were 1.50± 0.07 and 1.52 ± 0.12, respectively, which presented a tendency of synchronously increase after treated with ERK pathway activator EGF. The IC50 values in GEM-resistant cells of 0 nmol/L and 200 nmol/L levels were 4.104 and 10.20, respectively. After treated with U0126, these values decreased to 3.26 and 4.50, respectively; while treated with EGF, the IC50 values increased to 8.89 and 17.17, respectively. Conclusion： The ERK pathway may induce the GEM-chemoresistance in pancreatic cell line SW1990 through the participation in the regulation of the mdr-1 and RRM1 gene expression.
文摘BACKGROUND Insomnia is a disease where individuals cannot maintain a steady and stable sleep state or fail to fall asleep.Western medicine mainly uses sedatives and hypnotic drugs to treat insomnia,and long-term use is prone to drug resistance and other adverse reactions.Acupuncture has a good curative effect and unique advantages in the treatment of insomnia.AIM To explore the molecular mechanism of acupuncture at Back-Shu point for the treatment of insomnia.METHODS We first prepared a rat model of insomnia,and then carried out acupuncture for 7 consecutive days.After treatment,the sleep time and general behavior of the rats were determined.The Morris water maze test was used to assess the learning ability and spatial memory ability of the rats.The expression levels of inflammatory cytokines in serum and the hippocampus were detected by ELISA.qRTPCR was used to detect the mRNA expression changes in the ERK/NF-κB signaling pathway.Western blot and immunohistochemistry were carried out to evaluate the protein expression levels of RAF-1,MEK-2,ERK1/2 and NF-κB.RESULTS Acupuncture can prolong sleep duration,and improve mental state,activity,diet volume,learning ability and spatial memory.In addition,acupuncture increased the release of 1L-1β,1L-6 and TNF-αin serum and the hippocampus and inhibited the mRNA and protein expression of the ERK/NF-κB signaling pathway.CONCLUSION These findings suggest that acupuncture at Back-Shu point can inhibit the ERK/NF-κB signaling pathway and treat insomnia by increasing the release of inflammatory cytokines in the hippocampus.
基金supported by the Natural Science Foundation of Hubei Province(2024AFD252)the Fundamental Research Funds for the Central Universities South-Central Minzu University(Grant Numbers:CZZ24017)the Fundamental Research Funds for Health Commission of Hubei Province(ZY2023M022).
文摘This study aimed to evaluate the therapeutic properties of the traditional Chinese medicine Xuesanqi(XSQ,from the rhizome of Polygonum amplexicaule D.Don)in treating ulcerative colitis.We hypothesized that its many active components can alleviate symptoms of colitis by regulating the gut microbiota,its metabolites,and various signaling pathways.To test our hypotheses,we designed a DSS-induced colitis model in C57BL/6 male mice.Apparent metrics were evaluated in each group of mice and performed histological analysis of relevant tissues.The gut microbial composition was analyzed by 16S rRNA sequencing of bacteria.Simultaneously,the SCFAs content was detected by gas chromatography,inflammatory factor secretion was evaluated by ELISA or western-blot,the expression of tight junction protein and key proteins of the MAPK signaling pathway were analyzed by western-blot.Our result showed that the treatment with XSQ alleviated significant various symptoms such as weight loss,blood in stool,and shortening of colon.In addition,XSQ treatment restored the dysregulated gut microbiota in colitis mice,increased short chain fatty acids(SCFAs)and normalized the MAPK/ERK/JNK signaling pathways,promoted expression of tight junction protein Occludin,Claudin-1,and E-cadherin proteins.Furthermore,we also observed a dose-dependent pattern in these treatment responses.These findings demonstrated the active components of XSQ is a promising new treatment platform for ulcerative colitis.
文摘Objectives In this work,we explore the effect of atorvastatin on myocardial apoptosis and caspase-8 acti-vation after coronary microembolization(CME)in rats.Methods Fifty rats were randomly divided into five groups;the coronary microembolization(CME)group,the sham-operated(sham)control group,the gastric lavage control group,the atorvastatin lavage group,and the caspasse-8 inhibitor(N-acetyl-Ile-Glu-Thr-Asp-CHO,abbreviated as CHO)group,with 10 rats for each group.A microembolization ball was injected through the left ventricle for constructing the CME model.Animals in the sham control group were given an injection of physiological saline instead of the microembolization ball.Seven days before the operation,the atorvastatin group underwent gastric lavage with 20 mg/kg of atorvastatin once a day.Gastric lavage control animals underwent gastric lavage with an equivalent dose of physiological saline instead of the atorvastatin.Animals in the CHO group were given an intraperitoneal injection of 10 mg/kg of CHO 30 min before the operation.Six hours after the operation,cardiac ultrasonic detection was conducted on each group to measure the cardiac function indexes.TUNEL(Terminal-deoxynucleoitidyl transferase mediated dUTP nick end labeling)assays were used to measure myocardial apoptosis,and western blots were used to quantify the expression levels of activated caspase-3 and-8.Results(1)The echocardiographic parameters showed that,compared to the sham control animals,the left ventricular ejection fraction(LVEF)of the CME group was significantly decreased(P【0.05).In addition,cardiac sonography revealed a decrease in the left ventricular shortening fraction(FS)and cardiac output(CO),but an increase in the left ventricular end-diastolic dimension(LVEDd).Compared to the CME group,the atorvastatin and CHO groups exhibited significantly improved cardiac function(P【0.05).(2)When compared with the sham control,the myocardical apoptotic rate of the CME group,as well as the levels of activated caspase-3 and-8,increased significantly(P【0.05).The myocardial apoptotic rate,as well as the levels of activated caspase-3 and caspase-8 in the atorvastatin and CHO groups,decreased significantly(P【0.05)in comparison to the CME group.Conclusions The atorvastatin pretreatment clearly suppressed post-CME myocardial apoptosis and improved cardiac function.The most likely mechanism for these effects is the blockade of the myocardial death receptor-mediated apoptosis pathway.
基金Fund Project:Heilongjiang Natural Science Foundation Project(No.LH2020H104)Heilongjiang Postdoctoral Fund(No.LBH-Z20033)。
文摘Objective: To explore the therapeutic effect and underlying mechanism of Shenqi Zhilong Decoction on mice with membranous nephropathy (MN). Methods:Mice with MN was established by injecting cationic bovine serum albumin (c-BSA) into tail vein for several times. model mice were randomly divided into MN group (equal amount of distilled water), Shenqi Zhilong Decoction low dose group (12 g crude drug/kg), Shenqi Zhilong Decoction high dose group (24 g crude drug/kg), and Tripterygium wilfordii polyglycoside tablet group (14 mg/ kg). Another 10 un-treatment mice were taken as control group (equal amount of distilled water). The drug was administered orally once a day for 4 weeks. After the last administration, 24 hours urine was collected to determine the urinary protein content;blood from inner canthus was collected to measure the changes of kidney function, liver function, blood lipid and levels of IL-6, IL-4 and TNF-α in serum in each group;HE staining was used to observe the pathological changes of kidney. Immunohistochemical staining was used to observe the expression of IgG in kidney. The protein expression of ERK1/2 and cPLA2 in renal tissues was determined by Western-blot method. The gene expression of Neph1, Nephrin and Podocin mRNA in kidney tissues were detected by RT-PCR. Results: Compared with model group, Shenqi Zhilong decoction at low-dose and high-dose could significantly reduce the value of urine protein in MN mice;Decreased TC and TG levels (P<0.05 or P<0.01);Increased the levels of ALB and TP in liver function (P<0.05 or P<0.01);has no significant effects on the levels of CRE, UREA and UA in renal function (P>0.05). Decreased the contents of IL-6, IL-4 and TNF-α in serum (P<0.05 or P<0.01);Significantly down-regulated the protein expression levels of p-ERK1/2 and p-cPLA2 in kidney tissues of MN mice (P<0.05 or P<0.01);Significantly increased the expression levels of NephP1, Nephrin and Podocin mRNA in renal tissues (P<0.01). Conclusion: Shenqi Zhulong Decoction has a good therapeutic effect on MN mice, and the mechanism of action is related to regulate the expression of related genes of Nephrin-Podocin-Neph1 receptor complex for protecting the glomerular filtration barrier, and inhibite the activation of ERK/cPLA2 pathway for relieving damage of GEC and reduceing secretion of pro-inflammatory cytokines.
基金supported by the National Key Research and Development Project of China(No.2018YFA0800404)the National Natural Science Foundation of China(Nos.82100255 and 81970736)the China Postdoctoral Science Foundation(Nos.2021M691459 and 2022T150299).
文摘Cardiac fibrosis is a cause of morbidity and mortality in people with heart disease.Anti-fibrosis treatment is a significant therapy for heart disease,but there is still no thorough understanding of fibrotic mechanisms.This study was carried out to ascertain the functions of cytokine receptor-like factor 1(CRLF1)in cardiac fibrosis and clarify its regulatory mechanisms.We found that CRLF1 was expressed predominantly in cardiac fibroblasts.Its expression was up-regulated not only in a mouse heart fibrotic model induced by myocardial infarction,but also in mouse and human cardiac fibroblasts provoked by transforming growth factor-β1(TGF-β1).Gain-and loss-of-function experiments of CRLF1 were carried out in neonatal mice cardiac fibroblasts(NMCFs)with or without TGF-β1 stimulation.CRLF1 overexpression increased cell viability,collagen production,cell proliferation capacity,and myofibroblast transformation of NMCFs with or without TGF-β1 stimulation,while silencing of CRLF1 had the opposite effects.An inhibitor of the extracellular signal-regulated kinase 1/2(ERK1/2)signaling pathway and different inhibitors of TGF-β1 signaling cascades,comprising mothers against decapentaplegic homolog(SMAD)-dependent and SMAD-independent pathways,were applied to investigate the mechanisms involved.CRLF1 exerted its functions by activating the ERK1/2 signaling pathway.Furthermore,the SMAD-dependent pathway,not the SMAD-independent pathway,was responsible for CRLF1 up-regulation in NMCFs treated with TGF-β1.In summary,activation of the TGF-β1/SMAD signaling pathway in cardiac fibrosis increased CRLF1 expression.CRLF1 then aggravated cardiac fibrosis by activating the ERK1/2 signaling pathway.CRLF1 could become a novel potential target for intervention and remedy of cardiac fibrosis.
基金National Science Fund subsidized project(No.81774123)Key R&D Projects in Shaanxi Province(No.2018ZDXM-SF-011)。
文摘Objective:To observe the effect of Yishen Sanjie Huayu compound prescription on ERK/NF-κB signaling pathway in IgA nephropathy(IgAN)rats,and explore its effect on preventing and treating IgA nephropathy intrarenal arteriole disease.Methods:Fifty-five male SD rats were randomly divided into blank group,model group,ShenfukangⅡcapsule group and Losartan potassium tablet group.Bovine serum albumin(BSA)was used for intragastric administration and carbon tetrachloride(CCl4).IgAN rat model was established by subcutaneous injection and lipopolysaccharide(LPS)tail vein injection.ShenfukangⅡcapsule group and Losartan potassium tablet group were given each drug suspension 2ml/head/d one week after modeling Gavage was started.The blank group and the model group were given an equal volume of normal saline.The 24h urine protein(UTP)of the rats was measured at 4,8,and 12 weeks after the administration,and the blood creatinine(SCr)was measured after 12 weeks.,urea nitrogen(BUN),aldosterone(ADS),angiotensinⅡ(AngⅡ),immunohistochemical Envi-sion System two-step method to detect vascular endothelial growth factor(VEGF)and human matrix in the whole rat kidney and small artery area The expression of metalloproteinase-9(MMP-9),proliferating cell nuclear antigen(PCNA),extracellular regulatory protein kinase(ERK)1/2,nuclear transcription factor-κB(NF-κB),and the small arteries of rat kidney tissue The intima,media,vessel wall/vascular outer diameter value.Results:Compared with the model group,the expressions of VEGF,MMP-9,PCNA,ERK1/2 and NF-κB in kidney tissues of the ShenfukangⅡcapsule group and the Losartan potassium tablet group decreased(P<0.05),24hUTP and SCr,BUN level decreased(P<0.05),kidney tissue damage was alleviated;intima and vessel wall/vascular outer diameter values were significantly reduced(P<0.01),there was no significant difference in ADS between the groups.The AngⅡof the Tanpotassium tablets group was lower than that of the model group(P<0.05).Conclusion:Yishen Sanjie Huayu compound can inhibit the ERK/NF-κB signaling pathway in rats with IgA nephropathy,reduce the levels of VEGF,MMP-9,PCNA,ERK1/2,NF-κB,and inhibit intrarenal arteriole vascular endothelial cells Proliferate and reduce kidney damage.
基金funded by the grant for Evaluation of Functional Property,Processing and Utilization of Xinjiang Characteristic Plant Resources(G2023046003L)the Xinjiang Uygur Autonomous Region Outstanding Youth Science Fund project(2024D01E11)+1 种基金National Nature Science Fund of China(82060788)Hebei Natural Science Foundation(C2022204167).
文摘Coreopsis tinctoria Nutt.,an edible flowering plant,belongs to the Chrysanthemum family and is mainly grown at high altitudes in Northwestern China.It is rich in polyphenolic compounds,particularly marein and flavomarein,and possesses multiple health-promoting properties,such as antioxidant,hypoglycemic and vasorelaxant effects.Previous bioactivity investigations majorly focused on C.tinctoria and its crude extract.The aim of the present study was to prepare marein-dominant C.tinctoria flavonoids(CF),further investigate the CF protective effects of liver fibrosis induced by carbon tetrachloride and elucidate the associated molecular mechanisms.Results have demonstrated that CF effectively attenuated hepatofibrogenesis by increasing the activity of glutathione(GSH)and superoxide dismutase(SOD);suppressing the hepatic stellate cell(HSC)activation,inhibiting transforming growth factorβ(TGF-β)activation and the production ofα-smooth muscle actin(α-SMA),alleviating the phosphorylation of extracellular signal-regulated kinases1/2(ERK1/2)and small mothers against decapentaplegic1/2(Smad1/2),thus maintaining the collagen metabolic homeostasis in the liver.Our study revealed that CF possesses an efficacious protective effect against chronic hepatic fibrosis due to their strong inhibitory effects of oxidative stress and chronic inflammation.
基金supported by the National Natural Science Foundation of China(Nos.U22A20372,82204662,82374359,82204662)the Fujian Province Natural Science Foundation(Nos.2022J01369,2021J1942)+1 种基金the Developmental Fund of Chen Keji Integrative Medicine(No.CKJ2022010)the Fujian Natural Science Foundation for Distinguished Young Scholars(No.2024J010032)。
文摘Hypertension represents a significant chronic non-infectious disease in China,where Qingda Granule(QDG)has traditionally been employed for its management.However,the mechanisms underlying QDG's kidney protective effects remain incompletely understood.This study investigates QDG's role in ameliorating hypertensive kidney injury(KI)and elucidates the associated mechanisms.Network analysis identified potential therapeutic targets related to mitochondrial function and the extracellular signal-regulated kinase(ERK)cascade.Ribonucleic acid(RNA)sequencing revealed differentially expressed genes(DEGs)in hypertensive mouse kidneys,which were enriched in mitochondrial-related functions and normalized by QDG treatment.QDG attenuated angiotensin II(Ang II)-induced blood pressure elevation and enhanced renal artery flow.Both cellular and animal experiments demonstrated that QDG inhibits the ERK/ribosomal S6 kinase 1(RSK1)signaling axis,thereby preventing Ang II-induced mitochondrial damage and renal cell apoptosis.ERK pathway inhibitors confirmed QDG's mechanism of action through the ERK/RSK1 pathway.These findings indicate that QDG ameliorates hypertensive KI by preserving mitochondrial function through modulation of the ERK/RSK1 network,presenting a novel therapeutic approach for managing hypertensive KI in clinical practice.
基金supported by the National Natural Science Foundation of China(92374117,32170742,and 32100681)the National Key Research and Development Program of China(2025YFC2708200,2022YFC2702500,2025YFC2708001,and 2024YFC2706900)+2 种基金the Natural Science Foundation of Jiangsu Province(BK20220201)the State Key Laboratory of Reproductive Medicine and Offspring Health(SKLRM-2024B9)the Postgraduate Research&Practice Innovation Program of Jiangsu Province(KYCX24_2010 and KYCX24_2006)。
文摘The decline in oocyte quality and developmental potential with female reproductive aging is well recognized,yet the underlying mechanisms remain insufficiently investigated.In this study,an integrative analysis of transcriptomes and morphologies of individual oocytes from young and aged mice identifies morphologically defective aged oocytes with distinct transcriptomic features.Further analysis demonstrates that both apoptotic and ferroptotic pathways are activated in the defective aged oocytes,and simultaneously blocking both pathways reverses the defective morphology to the largest extent.The Plat gene,which encodes tissue-type plasminogen activator(t PA),is downregulated with oocyte aging,and Plat knockdown increases oocyte susceptibility to both apoptosis and ferroptosis.Mechanistically,t PA functions as an upstream signaling molecule for Erk1/2 activation by interacting with particular phosphorylation kinases such as Alk.Consequently,Plat loss downregulates Erk1/2 pathway activity in oocytes,leading to degeneration through PCD.Supplementing exogenous t PA in vitro oocyte maturation cultures reduces the defect rate of aged oocytes,thereby improving oocyte quality and developmental potential.Collectively,Plat plays a pivotal role in protecting aged mouse oocytes from PCD,and t PA supplementation may serve as a potential clinical strategy to enhance oocyte quality in females of advanced maternal age.
