雌二醇代谢产物对肝星状细胞增殖及TGF-β_1、CTGF表达的影响

The effects of estradiol metabolites on the proliferation and the production of TGF-β1 and CTGF of human hepatic stellate cells

目的研究雌二醇(E2)代谢产物:2-羟雌二醇(2OHE)、2-甲氧雌二醇(2MeOE)、4-羟雌二醇(4OHE)对活化的人肝星状细胞(LX-2)增殖及对细胞内转化生长因子β1(TGF-β1)和结缔组织生长因子(CTGF)水平的影响。方法用不同浓度的E2、2OHE、4OHE、2MeOE处理LX-2细胞48 h,实验终点用MTT法检测LX-2增殖情况、RT-PCR方法检测TGF-β1、CTGF mRNA的水平及细胞免疫化学法检测TGF-β1、CTGF蛋白的水平。结果在10-9M^10-7M的药物浓度范围,E2及E2代谢产物干预组的LX-2细胞光密度值(optical density,OD)和细胞内TGF-β1、CTGF mRNA及蛋白水平均较空白对照组低,E2代谢产物干预组各项指标下降更为明显,尤以2MeOE组突出,差异具有显著性(P<0.05)。结论 E2及E2代谢产物均能抑制肝星状细胞的增殖、下调TGF-β1和CTGF蛋白浓度,并呈剂量效应关系;比较同一浓度的不同干预药物,E2代谢产物的作用均强于E2,其中2MeOE的生物活性最强。

[ Objective ] To evaluate the effects of estradiol （E2） metabolites： 2-hydroxyestradiol （2OHE）, 4-hydroxyestradiol （4OHE） and 2-methoxyestradiol （2MeOE） on the proliferation and production of transforming growth factor-β,（TGF-β） and connective tissue growth faetor（CTGF） of activated human hepatic stellate cells（LX-2 cells）. [ Methods ] LX-2 cells were incubated with different concentrations of E2, 2OHE, 4OHE and 2MeOE for 48 hours, at the end point, analyzing the proliferation of the cells by MTr assay, detecting the mRNA and protein levels of TGF-β1, CTGF through RT-PCR and cellular immunochemistry separately. [Results] Among the concentrations of 10^-9 M-10^-7 M, these parameters such as optical density, mRNA levels and the concentrations of TGF-β1 and CTGF of LX-2 from the groups treated with estradiol or its metabolites were lower than those from the control group, and the values of the groups treated with estradiol metabolites were lower than the group treated with estradiol, especially in the group treated with 2MeOE, these difference had statistics significance （P 〈0.05）. [ Conclusions ] estradiol and its metabolites could inhibit the proliferation and production of transforming growth factor-β1 （TGF-β1） and connective tissue growth factor （CTGF） of hepatic stellate cells at a dose dependent manner, estradiol metabolites had stronger effect than E2 and 2MeOE had the highest activity.