外源性转化生长因子β_1对大鼠原代肝星状细胞活化的影响  

Effects of exogenous transforming growth factor-β_1 on the activation of primarily cultured rat hepatic stellate cells

作　　者：陈达凡[1] 李建英[1] 郑伟达[1] 陈治新[1] 王小众[1] 

机构地区：[1]福建医科大学附属协和医院消化内科,福建省福州市350001

出　　处：《世界华人消化杂志》2007年第3期211-215,共5页World Chinese Journal of Digestology

基　　金：福建省科技开发计划项目;No.2005D094~~

摘　　要：目的:探讨大鼠原代肝星状细胞(HSC)体外培养过程中,不同时期外源性转化生长因子β1(TGF-β1)对其活化的影响及相关因素的变化.方法:分离大鼠原代HSC,于无包被的塑料培养板上分别培养2,3,4,5,6,7d时予TGF-β15mg/L处理24h,倒置显微镜下观察细胞的形态变化,应用Western blot法检测处理前后细胞a-肌动蛋白(a-SMA)的变化,及活化过程中TGF-β1Ⅱ型受体(TβR-Ⅱ)的表达.结果:HSC在体外培养过程中,不同培养时间对TGF-β1的刺激活化作用反应不同.TGF-β1处理后,对培养2,3,4,5dHSC的活化有促进作用,以对培养第3天的细胞作用最明显,其a-SMA表达增加78.05%,而培养6,7d的HSC的形态和a-SMA变化不明显.培养第7天的HSC比培养第3天的细胞TβR-Ⅱ表达增高(3.30±0.83 vs 1.55±0.38,P<0.05).结论:TGF-β1对处部分活化状态中某阶段细胞的活化具有促进作用.完全活化的细胞对TGF-β1的刺激活化作用不敏感,原因与TβR-Ⅱ的表达量无关.AIM： To investigate the activation of primarily cultured hepatic stellate ceils （HSCs） at different differentiated stages after treatment of exogenous transforming growth factor-β （TGF-β1） and the alterations of related factors during this process. METHODS： HSCs were isolated from normal rats and primarily cultured in the uncoated plastics for 2, 3, 4, 5, 6, and 7 days. Then the ceils were incubated with 5 μg/L exogenous TGF-β1 for 24 hours. The morphological features of the cells were observed under inverted microscope. Western blot was used to detect the changes of or-smooth muscle actin （α-SMA） after TGF-β1 treatment and the expression of transforming growth factor β1 recepter-Ⅱ （TβR-Ⅱ ） during the activation of HSCs. RESULTS： During the process of in vitro cultivation, HSCs at different differentiated stages showed different responses to exogenous TGF-β1 treatment. TGF-β1 promoted the activation of HSCs after the cells had been cultured for 2, 3, 4, and 5 days. The cells cultured for 3 days were the most sensitive to TGF-β1, and the level of α-SMA expression was increasd by 78.05%. The changes of HSC morphology and α-SMA expression were not significant in the cells cultured for 6 and 7 days. TI3R-Ⅱ expression was significantly higher in HSCs cultured for 7 days than that in ones cultured for 3 days （3.30 ± 0.83 vs 1.55 ± 0.38, P 〈 0.05）. CONCLUSION： The activation of partiallyactivated HSCs can be promoted by exogenous TGF-β1, while the fuUy-activated HSCs lose this response. The amount of TβR-Ⅱ expression is not involved in this difference.

关 键 词：转化生长因子Β1 肝纤维化 肝星状细胞 Α肌动蛋白 

分 类 号：R575.2[医药卫生—消化系统]