骨形成发生蛋白对人骨髓间充质干细胞的影响

目的探讨骨形成发生蛋白(BMP)对人骨髓间充质干细胞的影响及调节作用.方法使用含BMP-7基因的PTracer-CMV载体感染人骨髓间充质干细胞(h MSCs),并设未转染组和空载体组,免疫组化法检测BMP-7蛋白表达,MTT法检测细胞增殖能力,流式细胞术检测细胞周期,湿化学法检测碱性磷酸酶合成情况.结果培养48 h后,BMP-7转染组h MSCs增殖速度明显高于未转染组和空载体组,差异具有统计学意义(P0.05);未转染组与空载体组h MSCs增殖速度之间比较差异无统计学意义(P0.05).BMP-7转染组各时间点G_0/G_1期细胞比例均明显低于未转染组和空载体组;S期、G_2/M期的细胞比例均明显高于未转染组和空载体组,上述差异具有统计学意义(P0.05);各时间点未转染组与空载体组G_0/G_1期、S期、G_2/M期细胞比例间差异均无统计学意义(P0.05).BMP-7转染组h MSCs细胞碱性磷酸酶含量明显高于未转染组、空载体组,差异具有统计学意义(P0.05).结论 BMP-7可促进h MSCs体外增殖和向成骨细胞分化,可能与促进细胞由G_1期进入S期、DNA合成增加、提升DNA合成的后期细胞数量有关.

Effect of Bone Morphogenetic protein on Human Bone Marrow Mesenchymal Stem Cells

Objective To investigate the influence and regulation effect of bone morphogenetic protein(BMP) on human bone marrow mesenchymal stem cells(MSCs). Method Human bone marrow mesenchymal stem cells(hMSCs)were infected by PTracer-CMV vector containing BMP-7 gene,and the untransfected group and empty vector group were also established. The expression of BMP-7 protein was detected by immunohistochemistry.Cell proliferation was detected by MTT,cell cycle was detected by flow cytometry,and the synthesis of alkaline phosphatase was detected by wet chemical method. Results After culturing 48 h,the increment rate of hMSCs in BMP-7 transfection group was significantly higher than that in the untransfected group and the empty carrier group(P<0.05).There was no significant difference in the speed of hMSCs increment between the untransfected group and the empty carrier group(P>0.05).The proportion of cells in G0/G1phase in BMP-7 transfection group at each time point were significantly lower than in the non-transfection group and empty vector group,while the cells in S phase and G2/M phase were significantly higher than those in the non-transfection group and empty vector group(P<0.05). The percentage of cells in G0/G1phase,S phase and G2/M phase at each time point showed no significant difference between the untransfection group and the empty vector group(P>0.05).The content of alkaline phosphatase in hMSCs cells of the BMP-7 transfected group was significantly higher than that in the untransfected group and the empty vector group,and the difference was statistically significant(P<0. 05). Conclusion BMP-7 can promote the proliferation and differentiation of hMSCs into osteoblasts in vitro.Its mechanism may be related to promoting cell proliferation from G1to S phase, increasing DNA synthesis and enhancing the number of cells at the later stage of DNA synthesis.