垂体中叶素抑制内质网应激拮抗阿霉素诱导的H9c2心肌细胞损伤机制研究

发布时间：2018-07-12 07:56

本文选题：阿霉素 + 心肌损伤　；参考：《广东医学》2017年12期

【摘要】：目的利用阿霉素(doxorubicin,DOX)诱导的H9c2心肌细胞损伤模型研究垂体中叶素(intermedin,IMD)拮抗DOX心脏损伤的药理学作用和机制。方法建立H9c2细胞的DOX心脏损伤模型,设空白对照组、DOX(2μmol/L)模型组、IMD受体拮抗剂IMD17-47(1μmol/L)组和IMD8-47(10~(-8)、10~(-6)和10~(-5)mol/L)3个剂量处理组,采用CCK-8法检测心肌细胞活性,丙二醛(MDA)含量测定评价H9c2细胞的氧化应激水平,Western法检测caspase-3活化片段和内质网应激相关蛋白葡萄糖调节蛋白78(GPR78)的表达水平,并采用Real-time PCR法检测内源性IMD及其受体系统的mRNA表达水平。结果与对照细胞相比较,DOX处理显著降低细胞存活率、增加细胞内MDA含量、caspase-3活性片段和内质网应激水平。与单纯DOX组相比较,IMD17-47抑制DOX诱导的内源性IMD的作用后明显降低细胞的存活率(P0.05)、增加细胞内MDA含量(P0.01)、caspase-3活化片段和内质网应激水平(P0.01);外源性给予IMD干预呈剂量依赖性地促进DOX处理细胞的存活率、降低细胞的MDA含量、凋亡和内质网应激水平(P0.05)。此外,DOX处理明显增加心脏组织内源性IMD及其受体的mRNA表达。结论 DOX诱导心肌细胞内源性IMD及其受体的表达,内源性IMD和外源性给予的IMD均通过DOX刺激增强的IMD受体系统,有效降低DOX诱导的氧化应激和内质网应激水平,实现其促进心肌细胞存活、抑制细胞凋亡的保护作用。
[Abstract]:Objective to study the pharmacological effect and mechanism of intermedin (IMD) on the cardiomyocyte injury induced by doxorubic indox (DOX). Methods the DOX heart injury model of H9c2 cells was established. The myocardial activity was measured by CCK-8 method in the dox (2 渭 mol / L) model group, IMD17-47 (1 渭 mol / L) and IMD8-47 (10 ~ (-8) -10 ~ (-6) and 10 ~ (-5) mol / L) groups. The level of oxidative stress in H9c2 cells was evaluated by measuring the content of malondialdehyde (MDA). The expression of caspase-3 activation fragment and endoplasmic reticulum stress-related protein glucose regulatory protein 78 (GPR78) was detected by Western blot. Real-time PCR was used to detect the mRNA expression of endogenous IMD and its receptor system. Results compared with control cells, DOX treatment significantly decreased cell survival rate, increased MDA content, caspase-3 activity fragment and endoplasmic reticulum stress level. Compared with DOX group, IMD17-47 significantly decreased cell survival rate (P0.05), increased intracellular MDA content (P0.01) and endoplasmic reticulum stress (P0.01) after inhibiting DOX-induced endogenous IMD (P0.01), and exogenous IMD intervention was dose-dependent. To promote the survival rate of cells treated with DOX, MDA content, apoptosis and endoplasmic reticulum stress were decreased (P0.05). In addition, DOX treatment significantly increased the expression of endogenous IMD and its receptor mRNA in cardiac tissue. Conclusion DOX induces the expression of endogenous IMD and its receptors in cardiomyocytes. Both endogenous IMD and exogenous IMD can effectively reduce the levels of oxidative stress and endoplasmic reticulum stress induced by DOX through the enhanced IMD receptor system stimulated by DOX. It can promote the survival of cardiomyocytes and inhibit the apoptosis of cardiomyocytes.
【作者单位】：四川大学;四川大学华西医院公共实验技术中心;
【基金】：国家自然科学基金资助项目(编号:31071001,31271226)
【分类号】：R96

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