全反式维甲酸对人胚肺成纤维细胞增殖及α-SMA表达的影响

Effect of all-trans-retinoic acid on the proliferation and expression ofα-SMA in human embryonic lung fibroblasts

目的:研究全反式维甲酸(ATRA)对人胚肺成纤维细胞(HFL-I)增殖与分化的影响。方法:体外培养HFL-I,MTT法检测不同浓度ATRA(0.1μmol/L、1μmol/L、10μmol/L)作用3d对HFL-I增殖能力的影响。5μg/L转化生长因子β_1(TGF-β_1)刺激0h、6h、12h、24h、48h、72h后,RT-PCR法检测α-平滑肌肌动蛋白(α-SMA)mRNA表达,刺激0d、1d、3d、5d后,Western blotting法检测α-SMA蛋白表达。不同浓度ATRA干预,24h后RT-PCR方法检测α-SMA mRNA表达,3d后用Western blotting方法检测α-SMA蛋白表达。结果:(1)MTT法检测显示不同浓度ATRA以浓度依赖性方式抑制HFL-I细胞的增殖(P<0.05)。(2)5μg/L TGF-β_1诱导后,HFL-I细胞中α-SMA mRNA和蛋白表达均上调(P<0.05)。(3)ATRA以浓度依赖性方式下调TGF-β_1诱导的α-SMA mRNA和蛋白的表达(P<0.05)。结论:ATRA能够抑制HFL-I细胞的增殖和TGF-β_1诱导的分化,该作用可能是通过下调α-SMA mRNA和蛋白的表达实现的。

AIM：To investigate the effects of all-trans-retinoic acid（ATRA）on the proliferation and differentiation of transforming growth factorβ_1（TGF-β_1）-stimulated human embryonic lung fibroblasts（HFL-I）. METHODS：The HFL-I cells were cultured in vitro and were pretreated with ATRA for 3 days at the concentrations of 0.1μmol/L,1μmol/L and 10μmol/L.The proliferation of HFL-1 cells was detected by MTT method.The mRNA expression ofα-smooth muscle actin（α-SMA）in HFL-I cells stimulated with TGF-β_1 for 0 h,6 h,12 h,24 h,48 h and 72 h was detected by RT-PCR and the protein expression ofα-SMA at the time points of 1,3 and 5 days was detected by Western blotting.The mRNA expression ofα-SMA in HFL-I cells pretreated with different concentrations of ATRA for 24 h was detected the by RT-PCR and the protein expression at time point of 3rd day was detected by Western blotting.RESULTS：Different concentration of ATRA inhibited the proliferation of HFL-I in a dose-dependent manner （P〈0.05）.Both mRNA and protein expression ofα-SMA in HFL-I cells pretreated with TGF-β_1 was up-regulated （P〈0.05）.ATRA down-regulated the mRNA and protein expression ofα-SMA induced by TGF-β_1 in a dose-dependent manner（P〈0.05）.CONCLUSION：ATRA inhibits the proliferation and TGF-β_1-stimulated differentiation in HFL-I cells by down-regulating the mRNA and protein expression ofα-SMA.