【摘要】 目的 探讨RhoA/ROCK2和NFκb信号传导通路在内脂素引起的心肌细胞肥大中的作用。方法 购买出生2～3 d的SD乳鼠，利用差速贴壁法提取原代心肌细胞并进行培养。培养48 h后换为无血清的培养基继续培养24 h,之后48 h改为无血清的培养基继续培养24 h后，根据实验设计分为不同的干预组。应用Real time-PCR、Western-Blot技术检测不同分组心肌细胞中BNPmRNA及蛋白表达情况，免疫细胞化学法观察NFκB核转位情况。ipp软件计算心肌细胞表面积，Western-Blot技术检测RhoA、ROCK2和IκB-α蛋白表达情况。结果 与正常对照组比较，10 ng/ml, 50 ng/ml和100 ng/ml内脂素预处理24 h的心肌细胞中，RhoA,ROCK2蛋白表达升高，IκB-α蛋白表达降低(P<0.05,P<0.01)。与正常对照组比较，100 ng/ml内脂素预处理24、48、72 h的心肌细胞中，RhoAROCK2蛋白表达升高，IκB-α表达降低(P<0.05,P<0.01)。SN50+内脂素、辛伐他汀+内脂素、Y27632+内脂素预处理48 h的心肌细胞表面积，与内脂素干预比较降低(P<0.05,P<0.01),与正常对照组比较升高(P<0.05,P<0.01)。SN50+内脂素、辛伐他汀+内脂素、Y27632+内脂素预处理48h的心肌细胞BNP mRNA和蛋白表达，与内脂素干预比较降低(P<0.05,P<0.01),与正常对照组比较升高(P<0.05,P<0.01)。与正常对照组比较，辛伐他汀、Y27632、SN50予干预组心肌细胞中ROCK2和IκB-α蛋白表达比较差异有统计学意义(P<0.01)。结论 内脂素可以通过RhoA/ROCK-NFκB信号传导通路引起心肌细胞肥大。更多还原

【Abstract】 Objective To investigate the role of RhoA/ROCK2 and NFκb signaling pathways in cardiomyocyte hypertrophy induced by visfatin.Methods The SD suckling rats(2～3 d of age) were used to extract primary cardiomyocytes to be cultured in vitro by differential adherence method.After 48-hour culture, the medium was changed to serum-free medium and the cells were cultured for 24 hours, then the cells were divided into different intervention groups according to the experimental design.Real time-PCR and Western-Blot techniques were used to detect the expression levels of BNP mRNA and protein in cardiomyocytes of different groups, and the nuclear translocation of NFκB was observed by immunocytochemistry.The ipp software was used to calculate the myocardial cell surface area, and Western-Blot technique was used to detect the expression levels of RhoA,ROCK2 and IκB-α protein.Results As compared with those in control group, the expression levels of RhoA,ROCK2 protein in 10 ng/ml, 50 ng/ml, and 100 ng/ml visfatin pretreatment groups(24 hours) were significantly increased, however, the expression levels of IκB-α protein were significantly decreased(P<0.05).As compared with those in control group, the expression levels of RhoA,ROCK2 protein in 100 ng/ml visfatin pretreatment groups(24 h, 48 h, 72 h) were significantly increased, however, the expression levels of IκB-α protein were significantly decreased(P<0.05 or P<0.01).Moreover the surface area of cardiomyocytes pretreated by SN50 + lipidin, simvastatin+visfatin, Y27632+visfatin for 48 h was decreased, as compared with that by simple visfatin intervention(P<0.05),however, which was significantly increased in control group(P<0.05).There were significant differences in the expression levels of ROCK2 and IκB-αprotein between control group and simvastatin, Y27632,SN50 intervenyion groups(P<0.01).Conclusion The visfatin can result in cardiomyocyte hypertrophy through the RhoA/ROCK-NFκB signaling pathway.更多还原