摘要
目的探讨桦木酸(BA)对高糖诱导的H9c2心肌细胞氧化应激损伤的保护作用及分子机制。方法将H9c2心肌细胞分为对照(Con)组(5.55 mmol/L葡萄糖)、高糖(HG)组(33 mmol/L葡萄糖)、HG+BA组(33 mmol/L葡萄糖+20μmol/L BA)、HG+BA+LY294002(HG+BA+LY)组(33 mmol/L葡萄糖+20μmol/L BA+10μmol/L LY294002)。CCK⁃8法检测细胞活力,流式细胞术检测细胞凋亡和活性氧簇(ROS)含量,黄嘌呤氧化酶法测定超氧化物歧化酶(SOD)活性,硫代巴比妥酸法测定丙二醛(MDA)含量,Western blot法检测蛋白激酶B(Akt)、磷酸化蛋白激酶B(p⁃Akt)、核因子E2相关因子2(Nrf2)、血红素氧合酶1(HO⁃1)和NADPH醌氧化还原酶1(NQO1)蛋白表达水平。结果与Con组比较,HG组细胞活力、SOD活性、p⁃Akt、Nrf2、HO⁃1和NQO1降低(P<0.05),MDA含量、细胞凋亡率和ROS生成升高(P<0.05)。与HG组比较,HG+BA组细胞活力、SOD活性、p⁃Akt、Nrf2、HO⁃1和NQO1升高(P<0.05),MDA含量、细胞凋亡率和ROS生成降低(P<0.05)。与HG+BA组比较,HG+BA+LY组细胞活力、SOD活性、p⁃Akt、Nrf2、HO⁃1和NQO1降低(P<0.05),MDA含量、细胞凋亡率和ROS生成升高(P<0.05)。结论BA可通过激活磷酸肌醇3激酶/Akt/Nrf2信号通路,上调NQO1和HO⁃1蛋白表达,减轻高糖诱导的H9c2心肌细胞氧化应激损伤。
Objective To investigate the protective effect of betulinic acid(BA)on oxidative stress injury induced by high glucose in H9c2 cells and its underlying mechanisms.Methods The H9c2 cells was divided into 4 groups:control group(5.55 mmol/L glucose),high glucose(HG)group(33 mmol/L glu⁃cose),HG+BA group(33 mmol/L glucose+20μmol/L BA),HG+BA+LY group(33 mmol/L glu⁃cose+20μmol/L BA+10μmol/L LY294002).The cell viability was detected by CCK⁃8 assay.Flow cy⁃tometry was used to determine apoptosis and ROS.SOD activity was analyzed by xanthine oxidase method.The levels of MDA were measured by thiobarbituric acid method.The protein levels of protein kinase B(Akt),p⁃Akt,Nrf2,HO⁃1 and NQO1 were tested by western blot.Results Cell viability,SOD activity,Akt phosphorylation level,Nrf2,HO⁃1 and NQO1 were decreased,while MDA content,apoptosis rate and ROS generation were increased in HG group than in Con group(P<0.05).Cell viability,SOD activity,Akt phosphorylation level,Nrf2,HO⁃1 and NQO1 were increased,while MDA content,apoptosis rate and ROS generation decreased in HG+BA group than in HG group(P<0.05).Cell viability,SOD activity,Akt phosphorylation level,Nrf2,HO⁃1 and NQO1 were decreased,while MDA content,apoptosis rate and ROS generation were increased in HG+BA+LY group than in HG+BA group(P<0.05).Conclusion BA could attenuate high glucose⁃induced oxidative stress injury by activating PI3K/Akt/Nrf2 signaling path⁃way and up⁃regulating the protein levels of HO⁃1 and NQO1.
作者
陈万宏
刘东伟
黄圣明
CHEN Wanhong;LIU Dongwei;HUANG Shengming(Department of Emergency,People’Hospital of Luohe,Luohe 462000,China)
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2019年第12期917-921,共5页
Chinese Journal of Diabetes
基金
河南省医学科技攻关计划项目（201702343）
