藏药复方冠心宁颗粒对缺氧复氧损伤大鼠心肌细胞的保护作用及机制研究  

Protective Effect and Mechanism of Guanxinning Granule on Rat Cardiomyocytes with Hypoxia and Reoxygenation Injury

作　　者：佳哇 东知多杰 多杰 马志良 李啟恩 JIA Wa;Dongzhi-Duojie;DUO Jie;MA Zhiliang;LI Qien(Qinghai Institute of Tibetan Medicine,Xining 810016,China;State Key Laboratory of Tibetan Medicine Research and Development,Xining 810016,China)

机构地区：[1]青海省藏医药研究院,青海西宁810016 [2]藏药新药开发国家重点实验室,青海西宁810016

出　　处：《高原科学研究》2022年第3期60-68,共9页Plateau Science Research

基　　金：青海省科学技术厅科技成果转化专项项目(2020-SF-130)。

摘　　要：目的:研究藏药复方冠心宁颗粒(Guan Xin Ning Granules,GXN)对缺氧复氧(Hypoxia and Reoxygenation,HR)损伤大鼠H9c2心肌细胞的保护作用及可能机制。方法:将大鼠心肌细胞在无血清的培养基中缺氧孵育4 h后再复氧孵育24 h,以建立心肌细胞缺氧复氧损伤模型,并随机分为对照组、模型组、卡托普利组(阳性对照组)和GXN组(0.5μg/mL、5μg/mL、20μg/mL)。采用CCK-8法测定细胞活力,试剂盒测定细胞内乳酸脱氢酶(LDH)含量、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、Na^(+)-K^(+)-ATPase和Ca^(2+)-ATPase的活性以及线粒体膜电位,流式细胞技术测定细胞凋亡率,实时荧光定量PCR法(qRT-PCR)测定局部粘着斑激酶(FAK)、磷脂酰肌醇激酶(PI3K)和蛋白激酶B(Akt)基因mRNA的表达情况。结果:与对照组相比,心肌细胞经过缺氧复氧处理后,细胞活力、细胞内LDH含量和SOD、GSH-Px、CAT、Na^(+)-K^(+)-ATPase、Ca^(2+)-ATPase的活性以及线粒体膜电位均显著下降(P<0.01),细胞凋亡率显著升高(P<0.01),FAK的mRNA表达显著下调(P<0.05);不同浓度的GXN干预后,细胞活力、细胞内LDH含量和SOD、GSH-Px、CAT、Na^(+)-K^(+)-ATPase、Ca^(2+)-ATPase活性以及线粒体膜电位较模型组均显著升高(P<0.05或0.01),细胞凋亡率显著下降(P<0.01),FAK、PI3K和Akt基因的mRNA表达显著上调(P<0.05或0.01)。结论:GXN对缺氧复氧损伤大鼠心肌细胞具有明显的保护作用,其机制可能与GXN抗氧化应激和抑制细胞凋亡有关。Objective:The purpose of this study is to investigate the protective effect and mechanism of Guanxin⁃ning Granules(GXN)on rat cardiomyocytes injured by hypoxiaandreoxygenation(HR).Methods:Rat cardiomyo⁃cytes in serum-free medium were exposed to hypoxia for 4 h and then reoxygenated for 24 h to establish the model of hypoxia reoxygenation injury of cardiomyocytes,and were randomly divided into control group,model group,captopril group(positive control group)and GXN group(0.5μg/mL,5μg/mL,20μg/mL).Cell viability was determined by CCK-8 assay.The content of lactate dehydrogenase(LDH),the activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),catalase(CAT),Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase,and mitochondrial membrane potential were determined by kits,respectively.The apoptosis rates of cardiomyocytes were determined by flow cytometry,and mRNA expressions of Focal adhesion kinase(FAK),phosphatidylinositol 3-kinase(PI3K)and protein kinase B(Akt)were determined by real-time quantitative PCR(qRT-PCR).Results:Compared with the control group,the cell viability,intracellular LDH content,activities of SOD,GSH-Px,CAT,Na^(+)-K^(+)-ATPase and Ca^(2+)-ATPase,and mitochondrial membrane potential of cardiomyocytes decreased significantly(P<0.01).Meanwhile,the apoptosis rates of cells were significantly increased(P<0.05),and mRNA expressions of FAK were significantly down-regulated(P<0.05)after hypoxiaandreoxygenation.After intervention of GXN with different concentrations,cell viability,intracellular LDH content,activities of SOD,GSH-Px,CAT,Na^(+)-K^(+)-ATPase,Ca^(2+)-ATPase activity and mitochondrial membrane potential were significantly higher than those in the model group(P<0.05).The apoptosis rates of cells were significantly decreased(P<0.05)and mRNA expressions of FAK,PI3K and Akt genes was significantly up-regulated(P<0.05 or 0.01)after GXN intervention compared with the model group.Conclusion:GXN has a significant protective effect on rat cardiomyocytes injured by hypoxia and reoxygenation,and the

关 键 词：藏药复方 冠心宁颗粒 大鼠心肌细胞 缺氧/复氧损伤 保护作用 

分 类 号：R965[医药卫生—药理学]