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

Effect of leptin on express of collagen type Ⅰ and α-SMA in hepatic satellate cell stimulated by acetaldehyde

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(100nmol/L)处理组及联合处理组(乙醛200μmol/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TGF-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TGF-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 α-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 subcuhured in vitro. HSCs were treated with acetaldehyde （200μmol/L）, leptin （100nmol/L） or combination with acetaldehyde （200μmol/L） and leptin （100nmol/L）. Then real-time PCR was performed to investigate the mRNA level of transforming growth factor-β1（TGF-β1） , collagen type Ⅰ and or-smooth muscle action （ α-SMA）, Western blot was performed to investigate the level of α-SMA, the contents of TGF-β1and collagen type Ⅰ in the supernatant fluid were measured by ELISA. Results： Compared with control group, the express level of TGF-β1 and collagen type I 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 α-SMA level between acetaldehyde-treated group and leptin-treated group compared with control group （P 〉 0. 05 ）, but combi- nation group was the highest level among four groups （ P 〈 0. 05 ）. Conclusion： Leptin can synergistically promote acetalde- hyde-induced expression of α-SMA in HSC, but can not up-regnlate acetaldehyde-induced expression of collagen type Ⅰ and TGF- 131. This data highlights the differential regulation of collagen type Ⅰ and α-SMA in activation of HSC.