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目的:研究上调Smad7表达对肝星形细胞(HSC)α1(Ⅰ)和α1(Ⅲ)前胶原基因转录的作用.方法:应用Fugene6介导Smad7质粒转染体外培养的HSC-T6细胞.继续培养48 h.同时使用实时定量聚合酶链式反应(Real time-PCR),逆转录酶链式反应(RT-PCR)方法检测Smad7质粒组和正常对照组、空载质粒对照组α(Ⅰ)和α1(Ⅲ)前胶原mRNA水平.结果:与正常对照组和空载质粒对照组相比,Smad7质粒转染HSC-T6细胞48h后,Smad7 mRNA水平显著增高(1.29±0.18 vs 0.11±0.02,0.13±0.02,均P<0.01),α1(Ⅰ)前胶原mRNA表达明显下降(0.10±0.01 vs 1.18±0.15.1.07±0.12,均P<0.01),但α1(Ⅲ)前胶原基因表达无明显改变(0.72±0.00 vs 0.70±0.01,0.75±0.0l,均P>0.05).结论:Smad7能明显抑制HSC细胞α1(Ⅰ)前胶原mRNA转录.
Objective: To study the effect of up-regulating Smad7 expression on α1 (Ⅰ) and α1 (Ⅲ) procollagen gene transcription in hepatic stellate cells (HSC) .Methods: HSC-T6 cells were transfected with Smad7 plasmid by Fugene6 and cultured in vitro 48 h respectively.At the same time, Smad7 plasmid group and normal control group were detected by real time-PCR and reverse transcriptase chain reaction (RT-PCR) α1 (Ⅲ) procollagen mRNA.Results: Compared with normal control group and empty plasmid control group, the Smad7 mRNA level was significantly increased in HSC-T6 cells transfected with Smad7 plasmid (1.29 ± 0.18 vs 0.11 ± 0.02, 0.13 ± 0.10, P <0.01), the expression of α1 (Ⅰ) procollagen mRNA was significantly decreased (0.10 ± 0.01 vs 1.18 ± 0.15.1.07 ± 0.12, all P <0.01) (0.72 ± 0.00 vs 0.70 ± 0.01,0.75 ± 0.0l, all P> 0.05) .Conclusion: Smad7 can significantly inhibit α1 (Ⅰ) procollagen mRNA transcription in HSC cells.