摘要
目的 探讨转化生长因子-β1（TGF-β1）对大鼠肌成纤维细胞（MFB）增殖及胞内Smad蛋白磷酸化的影响。方法 TGF-β1（0．33～81pmol／L）在不同作用时间（5～120min）的条件下诱导MFB细胞活化，采用免疫吸附和Westernblot方法观察TGF-β1对MFB细胞内pSmad2C、pSmad2L、pSmad3L及Smad2／3蛋白表达的影响；采用M1Tr方法观察TGF-β1对MFB细胞增殖的影响。结果 在0．33—9pmol／L浓度范围内，TGF-β1均呈浓度依赖性刺激MFB细胞内Smad2C、Smad2L、Smad3L磷酸化，以9pmol／L最明显，TGF-β1在27、81pmol／L浓度时，Smad2C、Smad2L、Smad3L磷酸化仍处于高水平状态，而各组Smad2／3蛋白表达无明显变化。在TGF-β1为9pmol／L浓度条件下，30—60min孵育时间对MFB细胞增殖无明显影响，但可呈时间依赖性刺激MFB细胞内Smad2C、Smad2L、Smad3L磷酸化，以30min最明显，TGF-β1在60、120min时间时，Smad2C、Smad2L、Smad3L磷酸化仍处于高水平状态，而各组Smad2／3蛋白表达量无明显变化。结论 TGF-β1诱导MFB细胞内Smads磷酸化先于刺激细胞增殖。
Objective To investigate the effects of TGF-β1 on the proliferation and Smad proteins phosphorylation in rat myofibroblast. Methods The activation of MFB was induced by TGF-β1 （0.33 - 81 pmol/L） in different time scopes（5 - 120 rain）. The expression levels of pSmad2C, pSmad2L, pSmad3L and Smad2/3 in activating MFB induced by TGF-β1 were investigated by immunoprecipitation and Western blotting method. The proliferation of activating MFB was determined by MTI＇ assay. Results TGF-β1（0.33 -9 pmol/L） stimulated Smad2C, Smad2L, Smad3L phosphorylation in a concentration dependent manner in MFB, and the optimal concentration of TGF-β1 was 9 pmol/L, the phosphorylation levels of these proteins were still high at the concentration of 27,81 pmol/L, while the expression levels of Smad2/3 had no difference between each group. At the concentration of TGF-β1 9 pmol/L and the time between 30 -60 rain coincubated with TGF-β1, no significant difference was observed in MFB proliferation, but phosphorylation Smad2C, Smad2L, Smad3L were mediated by TGF-β1 in a time dependent manner, and at the time of 30 rain was the most evident, the phosphorylation levels of these proteins were still high at the time of 60,120 rain, while the expression levels of Smad2/3 had no difference between each group. Conclusion These results suggest that TGF-β1 might promote Smad2C, Smad2L, Smad3L phosphorylation in MFB in a certain concentration scopes and time scopes, and induce the proliferation of MFB when stimulating time of TGF-β1 is prolonged. TGF-β1 might induce Smads phosphorylation firstly, then stimulate MFB proliferation.
出处
《安徽医科大学学报》
CAS
北大核心
2006年第5期499-502,共4页
Acta Universitatis Medicinalis Anhui
