转化生长因子-β1对大鼠骨髓间充质干细胞增殖及Slug表达的影响

Effects of transforming growth factor-β1 on rat bone marrow mesenchymal stem cell proliferation and Slug expression

目的检测转化生长因子-β1(transforming growth factor-β1,TGF-β1)对体外培养大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cell,BMMSCs)增殖及Slug表达的影响。方法采用密度梯度离心结合贴壁法分离、培养大鼠BMMSCs,用免疫组织化学方法对培养第3代的细胞进行鉴定。用MTT法检测不同浓度TGF-β1对细胞增殖的影响;免疫荧光和免疫印迹法检测TGF-β1处理前后Slug的表达情况。结果密度梯度离心结合贴壁法能有效分离、纯化大鼠BMMSCs,免疫组织化学法检测显示CD29、CD44表达阳性,而CD34、CD45表达阴性;低浓度TGF-β1对BMMSCs的增殖有促进作用,高浓度却抑制BMMSCs的增殖。TGF-β1处理24 h,Slug蛋白表达明显增强。结论一定浓度的TGF-β1可以促进BMMSCs的增殖,而且引起Slug蛋白增加。

Objective To detect the effect of transforming growth factor-β1（TGF-β1） on cell proliferation and Slug expression of bone marrow mesenchymal stem cells（BMMSCs）.Methods Density gradient centrifugation combined with adherence culture was used to isolate and culture rat BMMSCs.BMMSCs cultured to the 3rd passage were characterized by immunohistochemistry.The effects of TGF-β1 at different concentrations on cell proliferation were measured by MTT test.The expression of Slug protein was detected by immunofluorescence assay and Western blot.Results The rat BMMSCs were effectively isolated and purified by density gradient centrifugation combined with adherence culture.The results of immunohistochemical staining showed that the BMMSCs had positive expression of CD29 and CD44 while negative expression of CD34 and CD45.TGF-β1 in low concentration showed promoting effect on cell proliferation,but TGF-β1 in high concentration inhibited cell proliferation of the rat BMMSCs.The expression of Slug protein increased after treatment with TGF-β1 for 24 h.Conclusions TGF-β1 at a certain concentration can promote BMMSCs proliferation and expression of Slug protein.