红景天苷对H9c2心肌细胞损伤的保护作用及机制研究  

The protective mechanism of Salidroside in H9c2 myocardial cells injury

作　　者：尹敬[1] 田炜[1] 魏建强[2] 崔书霞[3] 喇孝瑾[1] 徐菁蔓[1] 王亚[1] 李继安[1] 

机构地区：[1]河北联合大学,河北唐山063000 [2]河北省唐山工人医院,063000 [3]中国人民解放军第285医院,河北邯郸056001

出　　处：《时珍国医国药》2014年第9期2107-2110,共4页Lishizhen Medicine and Materia Medica Research

基　　金：中国科技部中国-匈牙利政府间科技合作项目(No.2010.178.5-8)

摘　　要：目的观察红景天苷对H2O2诱导的H9c2心肌细胞氧化应激损伤的线粒体保护作用,并探讨其具体的信号转导机制。方法利用大鼠胚胎心脏组织来源的H9c2细胞建立H2O2损伤模型,分别给予100,75,50,25,1μmol/L的红景天苷预处理,MTT法测定细胞存活率,确定红景天苷的最适保护浓度;应用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,证明红景天苷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;同时用Western-blot法检测磷酸化Akt,磷酸化GSK-3β蛋白表达,初步探讨其具体信号转导机制。结果与H2O2组比较,红景天苷各浓度组细胞存活率显著提高(P<0.05);TMRE荧光减弱程度均明显减低,以浓度50μmol/L时作用最明显(P<0.05);红景天苷(50μmol/L)可显著上调Akt及GSK-3β的磷酸化水平(P<0.05)。结论红景天苷能够通过线粒体保护途径对抗H2O2诱导的H9c2细胞氧化应激损伤,且该保护作用可能是通过PI3K/Akt通路促进其下游因子GSK-3β的磷酸化,从而抑制mPTP的开放实现的。Objective To investigate the protective mechanism of salidroside in mitochondria of myocardial H9c2 cells oxidative stress injury induced by hydrogen peroxide. Methods To establish the hydrogen peroxide injury model in the rat embryonic heart tissue derived H9c2 cells,giving concentration for 100,75,50,25,1μmol/L salidroside pretreatment,MTT method to detect cell survival to determine the optimun protecting concentration of salidroside. Confocal microscopy imaging method by laser scanning is used to judge the mitochondrial permeability transition pore（mPTP） opening degree. By Western blot method,the expression of phosphorylated Akt and GSK-3β group were detected. Results Compared with the control group,salidroside groups increased fluorescence decrease degree were decreased significantly,and the concentration of 50μM significantly（P 0. 05）; Join the salidroside group cell survival rates were significantly increased（P 0. 05）; and significantly increase Akt and GSK-3β phosphorylation levels（P 0. 05）. Conclusion Salidroside in H9c2 cells induced by H2O2 oxidative stress injury has a protective effect,and this effect may be through the PI3 K/Akt pathway to promote its downstream factor phosphorylation of GSK-3β,make its inactivation,thus inhibiting myocardial cell protective open of mPTP.

关 键 词：红景天苷 过氧化氢 线粒体通透性转移孔 H9C2细胞 

分 类 号：R285.5[医药卫生—中药学]