Notch1/Hes1通路活化通过抑制氧化应激改善高糖诱导的心肌细胞肥大
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摘要
目的研究Notch1/Hes1信号通路活化对高糖诱导的心肌细胞肥大的影响,及其对氧化应激的作用。方法原代培养大鼠心肌细胞,采用图像分析法检测心肌细胞表面积,BCA试剂盒分析细胞总蛋白含量,采用RTq PCR和Western blot技术检测Notch1、Hes1、超氧化物歧化酶(SOD1)及诱导型一氧化氮合酶(iNOS)蛋白表达水平,相关试剂盒检测细胞匀浆中丙二醛(MDA)及一氧化氮(NO)含量。结果与对照组比较,高糖能明显诱导心肌细胞表面积及总蛋白含量的增加(P<0.05),心肌细胞中Notch1、Hes1蛋白表达量也明显降低(P<0.05),同时氧化应激产物MDA、NO明显升高(P<0.05),并且SOD1显著降低、iNOS显著升高(P<0.05);激活Notch1/Hes1信号通路能够显著降低高糖引起的心肌细胞肥大及氧化应激损伤(P<0.05),而Notch1干扰慢病毒可阻断Notch1对心肌细胞肥大及氧化应激的上述改善作用(P<0.05)。结论激活Notch1可以降低高糖诱导的心肌细胞肥大,其作用机制可能与降低细胞氧化应激损伤有关。
Aim To study the effect of Notch1/Hes1 pathway activation on the cardiomyocyte hypertrophy induced by high glucose(HG),and observe its role in oxidative stress.Methods Primarily cultured rat cardiomyocytes were used in the study.The cell surface areas of the cardiomyocytes were measured by an image analysis system.The cell protein content was detected by BCA method.The expression levels of Notch1,Hes1,superoxide dismutase 1(SOD1) and induction nitric oxide synthase(iNOS) were determined by RT-qPCR and Western blot.Related kits were used to measure the content of malondialdehyde(MDA) and mtric oxide(NO) in cardiomyocytes.Results Compared with control group,the cell surface areas and total protein content of the cardiomyocytes were significantly increased(P<0.05),the expression levels of Notch1 and Hes1 were significantly decreased(P<0.05),the levels of MDA,NO and iNOS were significantly increased(P<0.05),and the expression of SOD1 was greatly decreased(P<0.05) in HG-induced cardiomyocytes.Notch1 activation suppressed HG-induced cardiomyocyte hypertrophy and the levels of oxidative stress(P<0.05).These effects of Notch1 activation were abolished by Notch1 gene interference(P<0.05).Conclusion Notch1 activation reduces HG-induced cardiomyocyte hypertrophy by decreasing oxidative stress in cardiomyocyte.
Aim To study the effect of Notch1/Hes1 pathway activation on the cardiomyocyte hypertrophy induced by high glucose(HG),and observe its role in oxidative stress.Methods Primarily cultured rat cardiomyocytes were used in the study.The cell surface areas of the cardiomyocytes were measured by an image analysis system.The cell protein content was detected by BCA method.The expression levels of Notch1,Hes1,superoxide dismutase 1(SOD1) and induction nitric oxide synthase(iNOS) were determined by RT-qPCR and Western blot.Related kits were used to measure the content of malondialdehyde(MDA) and mtric oxide(NO) in cardiomyocytes.Results Compared with control group,the cell surface areas and total protein content of the cardiomyocytes were significantly increased(P<0.05),the expression levels of Notch1 and Hes1 were significantly decreased(P<0.05),the levels of MDA,NO and iNOS were significantly increased(P<0.05),and the expression of SOD1 was greatly decreased(P<0.05) in HG-induced cardiomyocytes.Notch1 activation suppressed HG-induced cardiomyocyte hypertrophy and the levels of oxidative stress(P<0.05).These effects of Notch1 activation were abolished by Notch1 gene interference(P<0.05).Conclusion Notch1 activation reduces HG-induced cardiomyocyte hypertrophy by decreasing oxidative stress in cardiomyocyte.
引文
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[15]Urbanek K,Cabral-da-Silva MC,Ide-Iwata N,et al.Inhibition of Notch1-dependent cardiomyogenesis leads to a dilated myopathy in the neonatal heart[J].Circ Res,2010,107:429-441.
[16]陈伟,陈嘉勤,毛海峰,等.有氧运动和黑果枸杞多糖对慢性脑缺血小鼠的干预及Notch通路相关因子的组织差异表达[J].中国动脉硬化杂志,2017,25(8):783-790.
[2]Chen R,Peng X,Du W,et al.Curcumin attenuates cardiomyocyte hypertrophy induced by high glucose and insulin via the PPARγ/Akt/NO signaling pathway[J].Diabetes Res Clin Pract,2015,108(2):235-242.
[3]Peng X,Chen R,Wu Y,et al.PPARγ-PI3K/AKT-NOsignal pathway is involved in cardiomyocyte hypertrophy induced by highglucose and insulin[J].J Diabetes Complications,2015,29(6):755-760.
[4]Yang K,Xu X,Nie L,et al.Indoxyl sulfate induces oxidative stress and hypertrophy in cardiomyocytes by inhibiting the AMPK/UCP2 signaling pathway[J].Toxicol Lett,2015,234(2):110-119.
[5]Nemir M,Metrich M,Plaisance I,et al.Control of cardiac hypertrophy and fibrosis by the Notch Pathway[J].Circulation,2011,124(14):116-119.
[6]Wang C,Li X,Wang H,et al.Notch1-nuclear factorκBinvolves in oxidative stress-induced alcoholic steatohepatitis[J].Alcohol Alcohol,2014,49(1):10-16.
[7]Pei H,Yu Q,Xue Q,et al.Notch1 cardioprotection in myocardial ischemia/reperfusion involves reduction of oxidative/nitrative stress[J].Basic Res Cardiol,2013,108(5):373-342.
[8]魏潇,雷建明,郭静文,等.白藜芦醇抑制高糖诱导的H9C2心肌细胞系肥大[J].基础医学与临床,2017,37(3):320-324.
[9]王涛,余志斌,谢满江,等.新生大鼠心肌细胞培养技巧[J].第四军医大学学报,2013,24(2):2-3.
[10]Adeghate E,Singh J.Structural changes in the myocardium during diabetes-induced cardiomyopathy[J].Heart Fail Rev,2014,19(1):15-23.
[11]Tham YK,Bernardo BC,Ooi JY,et al.Pathophysiology of cardiac hypertrophy and heart failure:signaling pathways and novel therapeutic targets[J].Arch Toxicol,2015,89(9):1401-1438.
[12]孟哲颖,王玉,林艳端,等.成纤维细胞生长因子21对高糖诱导心肌细胞肥大的保护作用[J].上海交通大学学报(医学版)[J].2015,35(1):23-28.
[13]Elnakish MT,Ahmed AA,Mohler PJ,et al.Role of oxidative stress in thyroid hormone-induced cardiomyocyte hypertrophy and associated cardiac dysfunction:an undisclosed story[J].Oxid Med Cell Longev,2015,12(9):854-867.
[14]叶挺,程治源,凌秋洋,等.Notch1蛋白在人心脏瓣膜间质细胞凋亡与钙化关系中的作用[J].中国动脉硬化杂志,2017,25(2):122-128.
[15]Urbanek K,Cabral-da-Silva MC,Ide-Iwata N,et al.Inhibition of Notch1-dependent cardiomyogenesis leads to a dilated myopathy in the neonatal heart[J].Circ Res,2010,107:429-441.
[16]陈伟,陈嘉勤,毛海峰,等.有氧运动和黑果枸杞多糖对慢性脑缺血小鼠的干预及Notch通路相关因子的组织差异表达[J].中国动脉硬化杂志,2017,25(8):783-790.