黄芪皂苷甲对转化生长因子β1刺激引起大鼠肾成纤维细胞氧化应激的影响  

Effect of astragaloside Ⅳ on transforming growth factor-β1 induced oxidative stress in renal fibroblasts of rats

作　　者：刘欣艳[1] 曹爱丽[1] 王浩[1] 王云满[1] 池杨峰[1] 韩文可 王利[1] 彭文[1] 

机构地区：[1]上海中医药大学附属普陀医院肾内科,200062

出　　处：《中华生物医学工程杂志》2015年第2期104-107,共4页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金项目（81473480）;国家中医药管理局重点学科建设项目（中医全科医学）;上海市医学重点专科建设项目（ZK2012A34）;上海市教委预算内项目（2012JW71）

摘　　要：目的：观察黄芪皂苷甲对转化生长因子β1（TGF-β1）刺激引起大鼠肾成纤维细胞氧化应激损伤的保护作用及其机制。方法体外培养大鼠正常肾成纤维细胞NRK-49F，实验分为6组，即对照组、TGF-β1刺激组、二甲基亚砜（DMSO）组、AS-Ⅳ10μmol/L组，AS-Ⅳ20μmol/L组，AS-Ⅳ40μmol/L组。除对照组外，其余5组均加入TGF-β1（终浓度1 ng/ml）复制细胞纤维化模型；AS-Ⅳ10、20、40μmol/L组分别给予相应剂量的AS-Ⅳ进行干预，DMSO组为阴性对照。检测不同浓度AS-Ⅳ对细胞增殖抑制率的及细胞存活率的影响，CM-H2DCFDA荧光探针测定细胞内活性氧簇（ROS）含量。结果与对照组比较，AS-Ⅳ浓度在40μmol/L以下时，细胞增殖抑制率无明显改变（均P〉0.05）。TGF-β1刺激组、AS-Ⅳ20μmol/L组、AS-Ⅳ40μmol/L组的细胞存活率分别为（68.33±1.29）%、（73.43±1.74）%、（86.9±2.94）%，与TGF-β1刺激组比较，AS-Ⅳ20、40μmol/L组细胞存活率均显著升高（均P〈0.01）。与对照组比较，TGF-β1刺激组细胞内ROS荧光强度明显增强，而加入10、20、40μmol/L AS-Ⅳ干预后，各组细胞荧光强度逐渐减弱，以AS-Ⅳ40μmol/L组减弱程度最强。结论 AS-Ⅳ可显著抑制TGF-β1诱导的细胞纤维化模型中细胞内ROS合成，提示其可能通过抑制氧化应激途径发挥抗纤维化作用。Objective To investigate the protective effects and underlying mechanism of astragalosideⅣ（AS-Ⅳ）on transforming growth factor-β1（TGF-β1）induced oxidative stress injury in renal fibroblasts of rats. Methods The normal renal fibroblasts NRK-49F of rats were cultured in vitro and divided into 6 groups,including the control group,TGF-β1 stimulation group,dimethyl sulfoxide（DMSO） group and AS-Ⅳ10,20,40μmol/L groups. Except for the control group,the cellular fibrosis models were duplicated in the rest five groups by adding TGF-β1（with the final concentration of 1 ng/ml）. The AS-Ⅳ10, 20 and 40 μmol/L groups was given the corresponding dose of AS-Ⅳ as an intervention,respectively. The DMSO group was negative control group. The effects of different concentrations of AS-Ⅳ on proliferation inhibiting rate and survival rate of NRK-49F were determined. The contents of reactive oxygen species （ROS）in NRK-49F were determined by CM-H2DCFDA fluorescent probe. Results Compared with the control group,no significant changes were found on the NRK-49F proliferation inhibiting rate when AS-Ⅳat and below 40μmol/L（all P〉0.05）. The NRK-49F survival rate in the TGF-β1 stimulation group,AS-Ⅳ20μmol/L group and AS-Ⅳ 40 μmol/L group was （68.33 ± 1.29）%,（73.43 ± 1.74）% and （86.9 ± 2.94）%,无nbsp;respectively. The NRK-49F survival rates in the AS-Ⅳ20 and 40μmol/L groups were significantly increased compared with that in the TGF-β1 stimulation group（both P〈0.01）. Compared with the control group,the intracellular fluorescence intensity of ROS in NRK-49F significantly increased in the TGF-β1 stimulation group,whereas gradually decreased in AS-Ⅳ10,20 and 40μmol/L groups,and decreased the most in AS-IV 40 μmol/L group. Conclusion AS-Ⅳ can significantly inhibit intracellular ROS synthesis in TGF-β1 induced cellular fibrosis model,indicating its anti-fibrotic mechanism via inhibiting oxidative stress.

关 键 词：黄芪皂苷甲 肾间质纤维化 转化生长因子Β1 氧化应激 

分 类 号：R575.2[医药卫生—消化系统]