黄芪3种成分对ChangLiver细胞氧化应激的抑制作用(1)
[摘要]本研究主要在于探讨黄芪3种成分(黄芪甲苷、毛蕊异黄酮葡萄糖苷、芒柄花素)对H2O2诱导ChangLiver细胞氧化应激的影响。以人正常肝实质细胞ChangLiver细胞为研究对象，同时选取具有明确保肝作用的联苯双酯为阳性对照药物，通过H2O2诱导氧化应激建立损伤模型进行实验。设立空白对照组、氧化应激组、黄芪甲苷组、毛蕊异黄酮葡萄糖苷组、芒柄花素组、阳性对照组。通过微板法、比色法检测内源性抗氧化体系相关指标，DCFH-DA荧光探针检测细胞内活性氧水平，分光光度法、Real-timePCR法、Westernblot法分别检测细胞肝微粒体CYP2E1的活性及其mRNA和蛋白的表达变化。从实验结果中可以看出，H2O2的诱导使ChangLiver细胞抗氧化能力降低，细胞内活性氧水平及CYP2E1表达的升高，而3种黄芪成分对细胞的预处理均可显著或极显著的改变上述氧化应激状态(与氧化应激组比，P<0.05，P<0.01)，总体效果与阳性对照组药物联苯双酯作用相当。由此表明，3种黄芪成分(黄芪甲苷、毛蕊异黄酮葡萄糖苷、芒柄花素)对H2O2诱导所致ChangLiver细胞氧化应激均有显著或极显著的抑制作用，细胞的氧化损伤得到明显缓解。
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[关键词]黄芪;ChangLiver细胞;H2O2;氧化应激
Inhibiting effects of three components of Astragalus membranaceus
on oxidative stress in Chang Liver cells
LI Jian， HAN Lin， MA Yu-fang， HUANG Yi-fan*
(University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal
Healthcare in Fujian Province， Fujian Agriculture and Forestry University， Fuzhou 350002， China)
, 百拇医药
[Abstract]The main objective of this research is to investigate the effects of astragaloside IV， calycosin separately glucoside， formononetin on oxidative stress in Chang Liver cells induced by H2O2. In the experiments， Chang Liver cells(a kind of normal human hepatocytes) were used as the research object， bifendate which has a clear hepatoprotective effect was used as the positive control drug， then the oxidative damage model of Chang Liver cells were established by H2O2. Cells were divided into six groups： blank control group，oxidative stress group， astragaloside IV group， calycosin separately glucoside group， formononetin group and positive control group. Then endogenous antioxidant system related indexes were detected by micro plate and colorimetric method; intracellular reactive oxygen species(ROS) were detected by DCFH-DA fluorescent probe; and the expressions of CYP2E1 were evaluated by liver microsomes， mRNA， and protein， respectively with spectrophotometry， Real-time PCR method， and Western blot technique. Results showed that H2O2 decreased antioxidant activity， and increased ROS level and expression of CYP2E1. The above oxidative stress status had been changed with protections of the three components of Astragalus membranaceus(compared with oxidative stress group， P<0.05，P<0.01)， which taken as a whole had equivalent effects as the drug of positive control group(bifendate). Taken together， three Astragalus membranaceus ingredients all had significant or extremely significant inhibiting effects on oxidative damaged Chang Liver cells which were induced by H2O2， and the oxidative damage of Chang Liver cells had been relieved., 百拇医药(李健 韩林 马玉芳 黄一帆)
, http://www.100md.com
[关键词]黄芪;ChangLiver细胞;H2O2;氧化应激
Inhibiting effects of three components of Astragalus membranaceus
on oxidative stress in Chang Liver cells
LI Jian， HAN Lin， MA Yu-fang， HUANG Yi-fan*
(University Key Laboratory for Integrated Chinese Traditional and Western Veterinary Medicine and Animal
Healthcare in Fujian Province， Fujian Agriculture and Forestry University， Fuzhou 350002， China)
, 百拇医药
[Abstract]The main objective of this research is to investigate the effects of astragaloside IV， calycosin separately glucoside， formononetin on oxidative stress in Chang Liver cells induced by H2O2. In the experiments， Chang Liver cells(a kind of normal human hepatocytes) were used as the research object， bifendate which has a clear hepatoprotective effect was used as the positive control drug， then the oxidative damage model of Chang Liver cells were established by H2O2. Cells were divided into six groups： blank control group，oxidative stress group， astragaloside IV group， calycosin separately glucoside group， formononetin group and positive control group. Then endogenous antioxidant system related indexes were detected by micro plate and colorimetric method; intracellular reactive oxygen species(ROS) were detected by DCFH-DA fluorescent probe; and the expressions of CYP2E1 were evaluated by liver microsomes， mRNA， and protein， respectively with spectrophotometry， Real-time PCR method， and Western blot technique. Results showed that H2O2 decreased antioxidant activity， and increased ROS level and expression of CYP2E1. The above oxidative stress status had been changed with protections of the three components of Astragalus membranaceus(compared with oxidative stress group， P<0.05，P<0.01)， which taken as a whole had equivalent effects as the drug of positive control group(bifendate). Taken together， three Astragalus membranaceus ingredients all had significant or extremely significant inhibiting effects on oxidative damaged Chang Liver cells which were induced by H2O2， and the oxidative damage of Chang Liver cells had been relieved., 百拇医药(李健 韩林 马玉芳 黄一帆)