甲基阿魏酸对TGF-β_1刺激人肝星状细胞增殖及活化的抑制作用

Inhibitory effect of meth-ferulic acid on proliferation and activation of TGF-β_1-induced human hepatic stellate cells

目的探讨甲基阿魏酸（MFA）对转化生长因子（TGF）-β_1刺激人肝星状细胞（HSC）-LX-2增殖及活化的抑制作用。方法将体外培养的HSC-LX-2细胞作为正常对照组,采用1μg/L TGF-β_1刺激HSC-LX-2进行细胞造模为模型组,药物组含终质量浓度为1μg/L的TGF-β_1和0.312 5、0.625、1.25、2.5、5、10、20、40 mg/L的MFA。根据MTT法检测MFA对药物组HSC-LX-2细胞增殖的影响,选择MFA作用浓度为1.25、2.5、5 mg/L作为低、中、高剂量组,将细胞孵育72h后,采用RT-PCR法检测各组α-平滑肌肌动蛋白（α-SMA）mRNA、Ⅰ型前胶原（PCⅠ）mRNA表达;采用Western blotting法检测各组α-SMA蛋白表达;采用ELISA法检测各组PCⅠ蛋白表达。结果在0.312 5-5 mg/L质量浓度范围内,随着MFA质量浓度升高,HSC-LX-2细胞生长抑制率逐渐升高,呈浓度依赖性（P均〈0.05）;当MFA质量浓度大于10 mg/L时,HSC-LX-2生长抑制率逐渐降低（P均〈0.05）。在1.25-5.0 mg/L范围内随MFA质量浓度逐渐增高,HSC-LX-2细胞中α-SMA及PCⅠ的mRNA及蛋白表达量逐渐降低（P均〈0.05）。结论 MFA可抑制TGF-β_1刺激的人肝星状细胞-LX-2增殖,减弱α-SMA、PCⅠ表达,抑制HSC-LX-2细胞外基质的合成及HSC-LX-2表型的转化。

Objective To investigate inhibitory effect of meth-ferulic acid（ MFA） on proliferation and activation of transforming growth factor-β_1（ TGF-β_1）-induced human hepatic stellate cells（ HSC-LX-2）. Methods HSC-LX-2 were cultured in vitro as the contol group and using 1 μg/L TGF-β_1to stimulate HSC-LX-2 as the model group. Meanwhile,the drug intervention group contained 1 μg/L TGF-β_1and 0. 312 5,0. 625,1. 25,2. 5,5,10,20 and 40 mg/L MFA. MTT assay was used to evaluate the effect of MFA on the proliferation of HSC-LX-2 in the model group and the drug intervention group,then the we chose the suitable concentrations of MFA into the low-dose,medium-dose and high-dose groups（ 1. 25,2. 5 and 5. 0 mg/L）. After 72 h incubation,the mRNA expression of α-SMA and PCⅠin the above five groups was determined by RT-PCR,the protein expression ofα-SMA was determined by Western blotting,and the protein expression of PCⅠin each group was determined by ELISA. Results With the increasing MFA concentration in the range of 0. 312 5-5 mg/L,the growth inhibition rate of HSC-LX-2 cell accelerated proportionally,which was in a dose-dependent manner（ all P〈0. 05）. However,when the MFA cell concentration was greater than 10 mg/L,the growth inhibition rate decreased. Similarly,within a certain concentration range（ 1. 25-5. 0 mg/L）,as MFA concentration increased,we found that the expression of α-SMA and PCⅠin HSC-LX-2 cells declined progressively（ all P〈0. 05）. Conclusion MFA may inhibit the proliferation,down-regulate the mRNA and protein expression of α- SMA and PCⅠin the TGF-β_1-induced HSC-LX-2 and inhibit the extracellular matrix synthesis and phenotype transformation of HSC-LX-2.