摘要
目的研究还原型谷胱甘肽(GSH)对人肝细胞和肝星状细胞(HSC)LX-2细胞株增殖和氧应激及转化生长因子(TGF)-β1表达的影响。方法分别用浓度为0.5~50 mmol/L和浓度为0.5~10 mmol/L的GSH培养肝细胞和HSC,MTT法检测GSH对肝细胞和HSC增殖的影响。用次氮基三乙酸铁(Fe-NTa)和GSH共同培养肝细胞和HSC,检测超氧化物歧化酶(SOD)活力和丙二醛(MDA)含量。用高(5.0 mmol/L)、中(2.5 mmol/L)、低(0.5 mmol/L)3个浓度的GSH培养HSC,酶联免疫吸附试验和荧光实时定量PCR方法检测TGF-β1蛋白和mRNA表达水平。结果在2.5~10 mmol/L浓度范围内,GSH可促进肝细胞增殖,各浓度GSH对HSC增殖均未见影响。GSH可增加肝细胞和HSC的超氧化物歧化酶活力,降低其丙二醛含量,同时可抑制HSC的TGF-β1表达。结论GSH可促进肝细胞增殖,保护肝细胞和HSC免受氧应激损伤,抑制HSC分泌TGF-β1,显示其具有肝细胞保护、抗氧化和抗肝纤维化作用。
Objective To investigate the impact of reduced glutathione(GSH) on the proliferation, oxidative stress and transforming growth factor β1(TGF-β1) expression of human hepatocytes and hepatic stellate cells(HSCs)(LX-2 cell line). Methods Human hepatocytes and HSCs were incubated with various concentrations of GSH(0.5-50 mmol/L or 0.5-10 retool/L). The effects of GSH on the proliferation of hepatocytes and HSCs were studied by 3-(4, 5 dimethylthiazol-2-yl)-2, 5-diphennylterazolium bromide colorimetric assay. Human hepatocytes and HSCs were co-cultured with GSH and ferric nitrilotriacetic acid, superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents were detected. HSCs were incubated with high(5.0 mmol/L), media(2. 5 mmol/L) and low (0.5 mmol/l.) concentrations of GSH, the expressions of TGF-β1 mRNA and protein were detected by ELISA and realtime PCR. Results In concentration ranged from 2. 5 to 10 mmol/L, the GSH could promote the proliferation of hepatocytes but no HSCs, significantly increased the activity of SOD and decrease the contents of MDA in hepatocytes and HSCs, and inhibited the expression of TGF-β1 in HSCs. Conclusions GSH can not only promote the proliferation of hepatocytes, but also protect hepatocytes and HSCs from oxidative stress, and inhibit the secretion of TGF-β1 in HSCs. GSH may play a role in hepatocellular protection, antioxidation and anti fibrosis.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2007年第10期666-669,共4页
Chinese Journal of Digestion
基金
上海市重点学科建设项目(Y0205)
上海市科学技术委员会基金(054009618)