外源Smad7基因对肝星状细胞Smad7 mRNA及蛋白表达的影响

目的观察外源Smad7基因能否有效转染肝星状细胞及其对Smad7 mRNA和蛋白表达的影响。方法构建鼠Smad7真核表达重组质粒，脂质体介导转染HSC—T6细胞，以RT—PCR和Western blot检测正常对照组、空质粒组及转染组中Smad7表达情况。结果Smad7真核表达质粒构建成功；外源Smad7体外转染肝星状细胞后，Smad7 mRNA和蛋白水平均显著上调，Smad7转染组与正常对照、空质粒组比较：Smad7 mRNA表达显著增加（P=0．009，0．011），蛋白水平显著上调（P=0．020，0．026），正常对照组、空质粒组Smad7 mRNA和蛋白水平表达差异无统计学意义（P=0．944，0．644）。结论Smad7真核表达质粒构建成功，外源Smad7基因可有效转染肝星状细胞，并显著上调Smad7 mRNA和蛋白水平。

Effect of exogenous Smad7 gene on expression of Smad7 in hepatic stellate cells

Objective To observe the influence of Smad7 transfection on endogenic expression of Smad7 in hepatic stellate cells. Methods Rat Smad7 eukaryotic ptasmid pcDNA3. 1 （ ＋ ）-Smad7 was constructed and transfected into HSC-T6 cells by Lipofectmine2000. The expression of Smad7 was determined by RT-PCR and Western blot respectively. Results Smad7 eukaryotic vector was successfully constructed and confirmed by enzyme digestion and sequencing. The levels of Smad7 mRNA and protein expression in Smad7 transfected group were significantly higher than control and empty vector group （P = 0. 009, 0.011 and P=0. 020, 0. 026）. The differences of Smad7 mRNA and protein expression were not statistically significant between control and empty vector group （P = 0. 944, 0. 664, respectively）. Conclusion Smad7 eukaryotic expression vector is successfully constructed which can express Smad7 effectively in HSC-T6 cells.