白藜芦醇对缺氧心肌细胞的保护作用及机制研究

发布时间：2018-08-26 06:51

【摘要】：目的:探讨白藜芦醇(resveratrol)是否可通过抑制内质网应激机制保护缺氧心肌细胞。方法:(1)H9c2心肌细胞随机分为对照组和缺氧组(1~8 h),Western blot法检测内质网应激(ERS)特征性分子伴侣葡萄糖调节蛋白78(GRP78)、葡萄糖调节蛋白94(GRP94)的表达以确定缺氧诱导ERS的时间。(2)培养H9c2心肌细胞建立缺氧模型,随机分为对照组、缺氧组、白藜芦醇+缺氧组、白藜芦醇组。Western blot检测缺氧对GRP 78,GRP 94,糖原合成酶激酶-3β(GSK-3β)的表达,观察白藜芦醇对缺氧所致GRP 78,GRP 94,GSK-3β磷酸化的影响;共聚焦显微镜观察白藜芦醇对缺氧所致线粒体通透性转移孔(mPTP)开放的影响;电镜观察白藜芦醇对缺氧所致心肌细胞超微结构的影响。结果:与对照组相比,缺氧3 h导致H9c2细胞GRP 78和GRP 94表达增加最多;白藜芦醇明显抑制缺氧所致GRP 78和GRP 94的表达及GSK-3β磷酸化水平。与对照组相比,缺氧组TMRE荧光强度明显减少,白藜芦醇明显抑制缺氧所致TMRE荧光强度的减少(即线粒体膜电位减少、mPTP开放)。与对照组相比,白藜芦醇明显减轻缺氧所致心肌细胞线粒体、内质网等超微结构的损伤。结论:白藜芦醇对缺氧所致H9c2心肌细胞损伤具有保护作用,该作用可能与抑制ERS使GSK-3β失活,进而阻止mPTP开放有关。
[Abstract]:Aim: to investigate whether resveratrol (resveratrol) can protect hypoxic cardiomyocytes by inhibiting endoplasmic reticulum stress. Methods: (1) H9c2 cardiomyocytes were randomly divided into two groups: control group and hypoxia group (1h). The expression of glucone-regulated protein 78 (GRP78) and glucose-regulated protein 94 (GRP94) in endoplasmic reticulum stress (ER) stress (ERS) were detected by Western blot to determine the time of hypoxia induced ERS. Hypoxia model was established by H9c2 cardiomyocytes. The rats were randomly divided into control group, hypoxia group, resveratrol hypoxia group and resveratrol group. Western blot was used to detect the expression of GRP 78 GRP94, glycogen synthase kinase 3 尾 (GSK-3 尾) and the effect of resveratrol on the phosphorylation of GSK-3 尾 in GRP 78 GRP94 induced by hypoxia. The effect of resveratrol on the opening of mitochondrial permeability transfer pore (mPTP) was observed by confocal microscope and the ultrastructure of cardiomyocytes induced by hypoxia was observed by electron microscope. Results: compared with the control group, the expression of GRP 78 and GRP 94 increased most in H9c2 cells after 3 h hypoxia, and resveratrol significantly inhibited the expression of GRP 78 and GRP 94 and the phosphorylation of GSK-3 尾. Compared with the control group, the fluorescence intensity of TMRE in hypoxia group was significantly decreased, and resveratrol significantly inhibited the decrease of TMRE fluorescence intensity induced by hypoxia (that is, the mitochondrial membrane potential decreased and TMRE opened). Compared with the control group, resveratrol significantly alleviated the damage of mitochondria, endoplasmic reticulum and other ultrastructure induced by hypoxia. Conclusion: resveratrol has a protective effect on H9c2 cardiomyocyte injury induced by hypoxia, which may be related to the inhibition of ERS inactivation of GSK-3 尾 and the prevention of mPTP opening.
【作者单位】：华北理工大学医学实验研究中心中匈合作中医药实验室唐山市新药基础研究重点实验室;
【基金】：国家自然科学基金(81570275) 河北省高等学校科学技术研究项目(ZD2014006,QN2014092) 华北理工大学杰出青年基金(JP201302) 大学生创新性实验计划项目(X2015046)
【分类号】：R54

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