天然产物研究与开发 ›› 2014, Vol. 26 ›› Issue (12): 2066-2070.

• 开发研究 • 上一篇    下一篇

通平养心方对高糖诱导H9c2心肌细胞损伤的保护作用机制研究

尹敬1,田炜1,魏建强2,喇孝瑾1,杨雨旸1,李继安1*   

  1. 1 河北联合大学 国家老年医学国际合作基地;2 唐山工人医院,唐山 063000
  • 出版日期:2014-12-31 发布日期:2015-01-09

Protective Mechanism of Tong Ping Heart Protection Formula on H9c2 Myocardial Cell Injury Induced by High Glucose

YIN Jing1 , TIAN Wei1, WEI Jian-qiang2, LA Xiao-jin1, YANG Yu-yang1, LI Ji-an1*   

  1. 1 Hebei United University,National Gerontology International Cooperation Base;2 Tangshan Worker Hospital,Tangshan 063000,China
  • Online:2014-12-31 Published:2015-01-09

摘要: 观察通平养心方对高糖诱导的H9c2细胞损伤的保护作用机制。高糖造成细胞损伤模型;MTT法检测细胞存活率;激光扫描共聚焦显微镜成像法,检测线粒体特异性荧光染料四甲基罗丹明乙酯(tetramethyrhodamine ester,TMRE),观察细胞线粒体膜电位(mitochondrial membrane potential,△Ψm)的变化,证明通平养心方是否通过抑制线粒体通透性转移孔(mitochondrial permeability transition pore,mPTP)的开放而发挥心肌线粒体的保护作用;Westernblot检测p-JNK、p-GSK-3β。结果发现,通平养心方和JNK抑制剂均能阻断高糖造成的H9c2心肌细胞损伤;0.1 μg/mL通平养心方预处理10 min,细胞存活率升高、TMRE荧光减弱程度降低,pJNK表达下降,p-GSK-3β表达升高。表明通平养心方能够通过线粒体保护途径对抗高糖诱导的H9c2细胞损伤,机制可能是通过JNK通路促进其下游因子GSK-3β磷酸化,从而抑制mPTP的开放实现的。

关键词: 糖尿病心肌病, 通平养心方, JNK通路, 心肌损伤

Abstract: To observe the protective mechanism of “Tong Ping heart protection” formula for H9c2 myocardial cell injured by high glucose.Cellular damage model was developed using high glucose;cell survival rate was detected by MTT method;the cell mitochondria membrane potential was detected by LSCM to identify whether “Tong Ping heart protection” formula protected myocardial mitochondria by inhibiting the mitochondrial permeability hole(mitochondrial permeability transition pore,mPTP)open;p-JNK and p-GSK-3β were tested by western blot.The results showed that “Tong Ping heart protection” formula and JNK inhibitors both can block H9c2 myocardial cell injury damaged by high glucose.The cell survival rate raised,TMRE fluorescence wanes reduced,p-JNK expression decreased and p-GSK-3β expression increased in the “Tong Ping heart protection” formula(0.1 μg/mL)pretreatment(10 min)group.These results indicated that “Tong Ping heart protection” formula can protect mitochondrial against H9c2 cell damage induced by high sugar.The protection mechanism was probably through promoting its downstream factor GSK-3β phosphorylation through JNK pathway,thus inhibiting mPTP open.

Key words: diabetic cardiomyopathy;“Tong Ping heart protection&rdquo, formula;JNK pathway;myocardial injury

中图分类号: 

R285.5