摘要
目的:研究蒙药尖叶假龙胆中雏菊叶龙胆酮对H2O2诱导的H9c2细胞氧化损伤的保护作用机制。方法:首先从尖叶假龙胆中提取雏菊叶龙胆酮单体化合物,建立H9c2细胞模型,将H9c2细胞分成五组:对照组,模型组,低、中、高剂量雏菊叶龙胆酮给药组(3.125、12.5、50μmol/L)。利用Real-time Cell Analysis(RTCA)多功能实时细胞功能分析仪建立最适过氧化氢损伤模型并检测活性。采用生化法检测细胞中超氧化物歧化酶(SOD)和乳酸脱氢酶(LDH)的活性,Western blot技术检测抑制剂作用前后血红素氧合酶-1(H0-1)、谷氨酰半胱氨酸连接酶催化亚基(GCLC)蛋白表达情况。结果:雏菊叶龙胆酮对H2O2诱导的H9c2细胞氧化损伤有一定的治疗作用。它能够减轻或抵消H2O2诱导的细胞损伤和LDH的释放,并增强SOD活力;H0-KGCLC蛋白的表达与模型组相比有明显的升高(P<0.05),BOS(GCLC抑制剂)和ZnPP(H0-1抑制剂)预处理显著降低雏菊叶龙胆酮的保护作用。结论:蒙药尖叶假龙胆中雏菊叶龙胆酮对H2O2诱导H9c2细胞损伤具有一定的保护作用,其机制可能是通过激活Nrf2/ARE信号通路下游的抗氧化蛋白酶HO-1、GCLC表达达到抗氧化保护目的。
Objective:To investigate the protective effect of bellidifolin from Mongolian medicine Gentianella acuta against H2O2-induced H9c2 cells injury.Methods:Firstly,the bellidifolin were extracted from Gentianella acuta,and then the H9c2 cells were divided into five groups:control group,model group,low,medium,high dose of bellidifolin groups(3.125、12.5、50μmol/L).The optimum concentration of hydrogen peroxide and the activity of bellidifolin were detected by Real Time Cellular Analysis(RTCA).The activities of superoxide dismutase(SOD)and lactate dehydrogenase(LDH)in H9c2 cells were measured by biochemical method.The expression of heme oxygenase-1(HO-l)and Glutamate-cysteine Iigase catalytic(GCLC)were evaluated using western blot.Results:Pre-treatment with different concentration of bellidifolin showed protection on H9c2 cells induced by H2O2 and it could improve SOD activity and inhibit LDH activity.Compared with model group,the expression of HO-l and GCLC protein in H9c2 cells were increased.Pretreatment with BOS(GCLC inhibitor)and ZnPP(HO-l inhibitor)significantly reduced the protective effect of bellidifolin.Conclusion:Bellidifolin,which was extracted from Gentianella acuta,has protective effect on H9c2 cells induced by H2O2,the mechanism may berelated to the expression of HO-l and GCLC regulated by Nrf2/HO-l pathway.
作者
任凯
龚雪
张瑞芬
张园
REN Kai;GONG Xue;ZHANG Rui-fen;ZHANG Yuan(Inner Mongolia Autonomous region Hospital of traditional Chinese Medicine,Hohhot 010020,China;Baotou Medical College,Baotou 014000,China;Inner Mongolia people’s hospital,Hohhot 010020,China)
出处
《中国民族医药杂志》
2020年第2期31-35,共5页
Journal of Medicine and Pharmacy of Chinese Minorities
基金
内蒙古自治区人民医院院内基金项目(No.2016040)。
