瘦素对乙醛诱导肝星状细胞活化过程中Ⅰ型胶原及α-平滑肌肌动蛋白表达的影响

摘要

目的:观察瘦素对乙醛诱导肝星状细胞( HSC)活化过程中α-SMA和Ⅰ型胶原表达的影响,旨在从体外探讨瘦素在酒精性肝病中的可能作用.方法:采用SD大鼠肝脏原位灌流消化的方法获得并原代及传代培养HSC,分乙醛(200μmol/L)处理组,瘦素(l00nmol/L)处理组及联合处理组(乙醛200μ mol/L加瘦素100nmol/L).用实时荧光定量PCR方法检测各组细胞TGF-β1、Ⅰ型胶原及α-SMA(α-平滑肌肌动蛋白)mRNA表达水平,Western blot检测各组细胞α-SMA表达量,ELISA法检测培养上清中TGF-β1及Ⅰ型胶原含量.结果:①乙醛处理组中TCF-β1和Ⅰ型胶原基因及蛋白表达量明显高于空白对照组(P＜0.05),α-SMA表达量与对照组相比差异无显著性意义(P＞0.05)；②瘦素处理组TGF-β1、Ⅰ型胶原及α-SMA表达量与对照组相比差异无显著性意义(P＞0.05)；③联合处理组TGF-β1、Ⅰ型胶原及α-SMA表达量明显高于对照组(P＜0.05),但TGr-β1、Ⅰ型胶原表达量与乙醛处理组相比差异无显著性意义(P＞0.05).结论:在乙醛诱导肝星状细胞活化过程中,瘦素能协同上调α-SMA的表达；但对TGF-β1及Ⅰ型胶原的表达无影响,Ⅰ型胶原和α-SMA的表达可能是依赖于不同的调控机制来实现.%Objective; To investigate the effect of leptin on the express of collagen type I and a-SMA in hepatic satellate cells (HSC) stimulated by acetaldehyde , to assess whether leptin contributes to alcoholic liver fibrosis by in vitro. Methods: HSCs were isolated from rats and then primarily cultured and subcultured in vitro. HSCs were treated with acetaldehyde (200μmol/L) , leptin (l00nmol/L) or combination with acetaldehyde (200μmol/L) and leptin (l00nmol/L). Then real-time PCR was performed to investigate the mRNA level of transforming growth factor-β1 (TGF-βl) , collagen type I and α-smooth muscle action ( α-SMA) , Western blot was performed to investigate the level of a-SMA, the contents of TCF-β1 and collagen type I in the supernatant fluid were measured by EUSA. Results: Compared with control group, the express level of TGF-β1 and collagen type 1 in acetaldehyde -treated group and in combination treatment group was significantly higher (P<0. 05), but acet-aldehyde-treated group was no significance compared with combination treatment group (P > 0.05). There was no significance at a-SMA level between acetaldehyde-treated group and leptin-treated group compared with control group (P > 0.05), but combination group was the highest level among four groups (P<0. 05). Conclusion; Leptin can synergistically promote acetalde-hyde-induced expression of a-SMA in HSC, but can not up-regulate acetaldehyde-induced expression of collagen type I and TGF-β1. This data highlights the differential regulation of collagen type I and a-SMA in activation of HSC.