P38 MAPK信号通路在漆树酸改善苯肾上腺素诱导的小鼠心肌细胞肥大中的作用

目的:探讨P38丝裂原活化蛋白激酶(P38 mitogen-activated protein kinase, P38 MAPK)信号通路在漆树酸改善苯肾上腺素(phenylephrine, PE)诱导的小鼠心肌细胞肥大中的作用。方法:原代培养新生小鼠心肌细胞,PE诱导心肌细胞肥大。按随机数字表法将细胞分为空白对照组、溶剂对照组、PE组、PE+漆树酸组、PE+漆树酸+P38抑制剂组和PE+P38抑制剂组。收集干预48 h后的乳鼠心肌细胞进行以下检测:Western blot检测p-P38、第9位赖氨酸乙酰化的组蛋白H3(acetylated histone H3 at lysine 9, H3K9ac)和心房钠尿肽(atrial natriuretic peptide, ANP)的蛋白表达水平,免疫共沉淀(co-immunoprecipitation, Co-IP)法验证p-P38与H3K9ac蛋白之间的相互作用,RT-qPCR检测肌细胞增强因子2C(myocyte enhancer factor 2C, MEF2C)mRNA表达水平,免疫荧光染色观察小鼠心肌细胞表面积。结果:West...

Effects of P38 MAPK signaling pathway on anacardic acid attenuating mouse cardiomyocyte hypertrophy induced by phenylephrine

AIM:To investigate the roles of P38 mitogen-activated protein kinase(P38 MAPK)in the process of anacardic acid(AA)attenuating mouse cardiomyocyte hypertrophy induced by phenylephrine(PE).METHODS:Cardiomyocyte hypertrophy was induced by PE in primary neonatal mouse myocardial cells.According to random number table method,the experiments were designed in 6 groups as following:control group,PE+DMSO group,PE group,PE+AA group,PE+AA+P38 inhibitor group and PE+P38 inhibitor group.Mouse myocardial cells were collected after intervention for 48 h.The protein levels of p-P38,acetylated histone H3 at lysine 9(H3K9ac)and atrial natriuretic peptide(ANP)were determined by Western blot.The interaction between p-P38 and H3K9ac was verified by co-immunoprecipitation(Co-IP).The mRNA expression of myocyte enhancer factor 2C(MEF2C)were tested by RT-qPCR.Mouse myocardial cell surface area was observed by immunofluorescence staining.RESULTS:The results of Western blot showed that the protein levels of p-P38 and H3K9ac in PE group were significantly increased compared with control group(P<0.05),and the levels of MEF2C mRNA and ANP protein in PE group were apparently increased compared with control group(P<0.05).However,P38 inhibitor and histone acetylase inhibitor AA attenuated P38 phosphorylation and H3K9 acetylation induced by PE,and down-regulated the over-expression of MEF2C and ANP in the mouse myocardial cells(P<0.05).The results of immunofluorescence staining showed that PE apparently increased mouse myocardial cell surface area(P<0.05),while P38 inhibitor and AA attenuated cardiomyocyte hypertrophy induced by PE(P<0.05).CONCLUSION:The mechanism of AA attenuating cardiomyocyte hypertrophy induced by PE may be related to the regulation of histone acetylation modification imbalance mediated by P38 MAPK signaling pathway.