荭草花提取物对H9c2心肌细胞缺氧复氧损伤的保护作用  

Protective Effect of Polygonum orientale Flower Extract on Cultured H9c2 Cardiomyocytes During Hypoxia/Reoxygenation Injury

作　　者：刘亭[1] 吴琼[1] 刘香香 陆定艳 薛维娜[1] 兰燕宇[1] 李月婷[1] LIU Ting;WU Qiong;LIU Xiang-xiang;LU Ding-yan;XUE Wei-na;LAN Yan-yu;LI Yue-ting(Guizhou Medcial University,Guiyang 550025,China)

机构地区：[1]贵州医科大学,贵州贵阳550025

出　　处：《中药材》2018年第1期184-188,共5页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金(81560630;81760699);贵州省科学技术厅人才团队项目(黔科合平台人才[2016]5613/5677);贵州省教育厅项目(黔教合协同创新字[2013]04)

摘　　要：目的:探究荭草花提取物对H9c2心肌细胞缺氧复氧损伤的保护作用。方法:利用氯化钴(CoCl_2)建立缺氧复氧损伤H9c2细胞模型,实验分正常对照组、模型组及荭草花提取物不同浓度组。应用MTS检测细胞存活率;生化试剂盒检测细胞乳酸脱氢酶(LDH)、肌酸激酶(CK)释放量及细胞内丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)水平;Annexin V-FITC/PI双染法流式细胞仪检测细胞凋亡情况;Western blot测定cleaved Caspase-3、Bcl-2、Bax蛋白表达。结果:利用800μmol/L CoCl_2缺氧22 h,复氧2 h可建立缺氧复氧损伤心肌细胞模型。与模型组比较,荭草花提取物能显著升高心肌细胞存活率,降低LDH、CK释放量及细胞内MDA含量,提高细胞内SOD、CAT活性,并呈浓度依赖性抑制CoCl_2诱导的缺氧复氧损伤。荭草花提取物能下调促凋亡蛋白cleaved Caspase-3、Bax的表达,上调凋亡抑制蛋白Bcl-2的表达,减少心肌细胞凋亡。结论:荭草花提取物对心肌细胞缺氧复氧损伤具有一定的保护作用,其机制可能与增强细胞清除自由基能力及抑制细胞凋亡有关。Obiective :To investigate the protective effects of Polygonum orientale flower extract on cultured H9c2 cardiomyoeytes during Hypoxia/Reoxygenation(H/R)Injury.Methods:CoC12 was used to induce the H/R injury model in H9c2 cardiacmyocytes at different concentrations.H9c2 cardiacmyocytes were randomly divided into the control group,model group and the Polygonum orientale flower extract different dosage groups.Myocardial cell viability was detected by the MTS assay kit.Lactate dehydrogenase (LDH),Creatine Kinase (CK),Malondialdehyde (MDA)content,Superoxidedimutase (SOD)and catalase (CAT)level were detected by biochemical kits.The apoptosis rate was detected by Annexin V-FITC/PI Flow-cytometry.The expressions of cleaved caspase-3,Bcl-2,Bax protein were detected by Western blot.Resuhs :800μmol/L CoC12 was used to hypoxia the cells for 22h and then reoxygenation for 2h to set up H/R injury model.Compared with the model group,Polygonum orientale flower extract can significantly increase the survival rate of myocardial cell,reduce the release of LDH,CK and MDA content,increase the activity of SOD and CAT,and hypoxia and reoxygenation injury induced by CoC12 was inhibited in a concentration dependent manner.Polygonum orientale flower extract depressed myocardial apoptosis by down-regulating the expression level of pro-apoptotic protein cleaved Caspase-3 and Bax,whereas up-regulating the expression of apoptosis inhibitory protein Bcl-2,and decreased the cardiomyosyte apoptosis.Conclusion:Polygonum orientale flower extract has a protective effect on CoC12 induced injury in H9c2 cells,the mechanism may be related to enhancing the ability of cell to scavenge free radicals and inhibiting apoptosis.

关 键 词：荭草花提取物 缺氧复氧 H9C2心肌细胞 

分 类 号：R285.5[医药卫生—中药学]