荭草花醇提物对H2O2诱导的H9c2细胞氧化损伤的保护作用机制研究  

Protective Effect of Flowers of Polygonum orientale Flower Ethanol Extract on H_2O_2-induced Oxidative Stress Injury in H9c2 Cells

作　　者：刘亭[1] 吴琼[1,3] 刘香香 陆定艳 黄文炫 李月婷 李勇军[2] LIU Ting;WU Qiong;LIU Xiang-xiang;LU Ding-yan;HUANG Wen-xuan;LI Yue-ting;LI Yong-jun(Provincial Key Laboratory of Pharmaceutics in Guizhou Province,Guizhou Medical University;Engineering Research Center for the Development and Application of Ethnic Medicine and TCM,Ministry of Education, Guizhou Medical University,Guiyang 550004,Guizhou;College of Pharmacy,Guizhou Medical University,Guiyang 550025,Guizhou)

机构地区：[1]贵州医科大学贵州省药物制剂重点实验室 [2]贵州医科大学,民族药与中药开发应用教育部工程研究中心,贵阳550004 [3]贵州医科大学药学院,贵州贵阳550025

出　　处：《天然产物研究与开发》2018年第2期299-303,共5页Natural Product Research and Development

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(2016-5613-5677);贵州省教育厅项目(2013-04)

摘　　要：研究荭草花醇提物对H9c2心肌细胞氧化损伤的保护作用机制。采用200μmol/L H_2O_2作用H9c2细胞0.5 h,建立H9c2细胞氧化损伤模型。将细胞分为正常对照组、H9c2细胞氧化损伤模型组、不同浓度荭草花醇提物(20、40、80μg/m L)预处理组。采用MTS法检测细胞存活率;生化试剂盒检测细胞乳酸脱氢酶(LDH)释放量,细胞内丙二醛(MDA)含量及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活力;Western blot测定cleavedcaspase-3、Bcl-2、Bax、p-AKT和AKT的表达。与模型组比较,荭草花醇提物能明显增加心肌细胞存活率,降低LDH释放量和MDA含量,提高SOD及CAT活性,并呈剂量依赖性抑制H_2O_2诱导的氧化应激损伤。荭草花醇提物下调cleaved caspase-3和Bax的蛋白水平,上调Bcl-2的表达,减少心肌细胞凋亡;增加细胞中p-AKT的表达,且这种表达可被PI3Ks抑制剂LY294002抵消。表明荭草花醇提物可减轻H_2O_2诱导的H9c2心肌细胞氧化应激损伤,其机制可能与减少细胞凋亡,平衡氧化应激产物有关,且部分依赖于磷脂酰肌醇3-激酶(PI3K/Akt)通路。To investigate the protective effect of Polygonum orientale Flower Ethanol Extracton H 2O 2 induced Oxidative damage in H9c2 Cells.200μmol/L H 2O 2 was used to induce the oxidativestress damage model in H9c2 cells for 0.5 h in vitro.The H9c2 cells were randomly divided into control group,the model group(200μmol/L H 2O 2)and the Polygonum orientale Flower ethanol Extract different groups(20、40、80μg/mL).Cellviability was determined by MTS assay.Thecontentoflactate dehydrogenase(LDH),malondialdehyde(MDA)and the activities of superoxidedimutase(SOD)and catalase(CAT)were detected by biochemical kits.The protein levels of caspase-3,Bax,Bcl-2,p-AKT and AKT were detected by Western blot.Compared with model group,Polygonum orientale Flower Ethanol Extract pre-treatment improved cell viabilityand the activities of SOD and CAT in a dose-dependent manner,and attenuated leakage of LDH and content of MDA.Polygonum orientale Flower Ethanol Extract depressed myocardial apoptosis by down-regulating pro-apoptotic protein cleavedcaspase-3 and Bax,up-regulating apoptosis inhibitory protein Bcl-2 and increasing the protein level of p-AKT. LY294002 could offsets such an effect of Polygonum orientale Flower Ethanol Extract.In summary,Polygonum orientale Flower Ethanol Extract showed a protective effect on H 2O 2 induced injury in H9c2 cells.This protection may result from inhibiting myocardial oxidative apoptosis,balancing the oxidative stress and maybe related with the PI3K/Akt signaling pathway.

关 键 词：荭草花醇提物 氧化应激损伤 H9C2心肌细胞 PI3K/Akt 

分 类 号：R96[医药卫生—药理学]