摘要
目的:探讨发动蛋白相关蛋白1(Drp1)在高糖诱导的心肌细胞肥大模型中的表达及其作用机制。方法:原代乳大鼠心肌细胞培养,用高糖诱导建立心肌细胞损伤模型,将细胞分为对照组、高渗透压组、DMSO组、高糖组和高糖+线粒体分裂抑制剂1(Mdivi-1)组。用麦胚凝集素荧光染色检测单个心肌细胞表面积;用线粒体膜电位(MMP)检测试剂盒(JC-1)检测心肌细胞MMP水平;Western blot检测心肌细胞肥大标志物心房钠尿肽(ANP)及Drp1和Drp1在Ser616/Ser637位点的磷酸化蛋白[p-Drp1(Ser616/Ser637)]的蛋白水平。结果:与对照组相比,高渗透压组各检测结果均无显著差异(P>0.05)。与对照组和高渗透压组相比,高糖组心肌细胞的表面积均显著增大(P<0.01),MMP显著降低(P<0.01),而Drp1总蛋白表达无显著变化(P>0.05);心肌细胞ANP表达显著增加(P<0.01),p-Drp1(Ser616)水平显著升高(P<0.05),而p-Drp1(Ser637)水平显著降低(P<0.05)。与高糖组相比,高糖+Mdivi-1组ANP和p-Drp1(Ser616)蛋白水平显著降低(P<0.05),而p-Drp1(Ser637)的水平显著升高(P<0.05)。结论:高糖诱导心肌细胞肥大的机制,与Drp1在Ser616位点磷酸化水平升高、Ser637位点磷酸化水平降低导致的线粒体分裂增加有关。
AIM:To investigate the effect of dynamin-related protein 1(Drp1)on high glucose-induced cardiomyocyte hypertrophy.METHODS:The primarily cultured rat cardiomyocytes were divided into normal glucose group,mannitol group,DMSO group,high glucose group and high glucose+mitochondrial division inhibitor-1(Mdivi-1)group.The area of single cardiomyocyte was calculated by wheat germ agglutinin staining.The mitochondrial membrane potential of the cardiomyocytes was detected by JC-1 staining,and the expression levels of atrial natriuretic peptide(ANP),the marker of cardiac hypertrophy,Drp1 and its phosphorylated proteins p-Drp1(Ser616)and p-Drp1(Ser637)were determined by Western blot.RESULTS:Compared with normal glucose group and mannitol group,the cell area was increased significantly and the mitochondrial membrane potential was decreased significantly in high glucose group.The protein levels of ANP and p-Drp1(Ser616)were significantly increased,the protein level of p-Drp1(Ser637)was decreased,and no significant difference of total Drp1 level among groups was observed.In high glucose+Mdivi-1 group,ANP expression and the protein level of p-Drp1(Ser616)were significantly decreased,and the protein level of p-Drp1(Ser637)was increased as compared with high glucose group.CONCLUSION:High glucose induces hypertrophy of cardiomyocytes,and the mechanism is associated with increased mitochondrial fission due to changes of Drp1 phosphorylation levels.
作者
吴清蕊
练飞鸿
饶芳
邝素娟
杨慧
吴飞龙
张梦珍
麦丽萍
林秋雄
单志新
杨敏
邓春玉
WU Qing-rui;LIAN Fei-hong;RAO Fang;KUANG Su-juan;YANG Hui;WU Fei-long;ZHANG Meng-zhen;MAI Li-ping;LIN Qiu-xiong;SHAN Zhi-xin;YANG Min;DENG Chun-yu(School of Biological Science and Engineering,South China University of Technology,Guangzhou 510006,China;Key Laboratory of Clinical Pharmacology,Guangdong Cardiovascular Institute,Guangdong General Hospital Medical Research Center,Guangdong Academy of Medical Sciences,Guangzhou 510080,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第1期47-52,共6页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.81470440)
关键词
糖尿病心肌病
发动蛋白相关蛋白1
线粒体分裂
心房钠尿肽
Diabetic cardiomyopathy
Dynamin-related protein 1
Mitochondrial fission
Atrial natriuretic peptide
