Follistatin表达抑制促进BMP-2诱导人类骨髓间充质干细胞成骨分化的实验研究

Experimental study on BMP-2-induced osteogenic differentiation of human bone marrow derived mesenchymal stem cells promoted by Follistatin depression

目的探讨Follistatin表达抑制对人类骨髓间充质干细胞(BMSC)细胞活性及骨形态发生蛋白(BMP)-2诱导人类BMSC成骨分化的影响。方法采用Follistatin特异性siRNA转染人类BMSC,检测线粒体脱氢酶活性、细胞DNA含量及蛋白含量。采用定量逆转录-聚合酶链式反应(qRT-PCR)检测成骨相关基因表达,采用碱性磷酸酶(ALP)染色和茜素红染色检测Follistatin对BMP-2诱导人类BMSC成骨分化的影响。结果转染后3、7 d,人类BMSC代谢、DNA含量显著升高;转染后14 d,总蛋白含量增加。Follistatin表达抑制后成骨相关基因如ALP、OCN、OSX、OC、OPN、RUNX2等表达升高,且ALP活性及钙沉积量显著增加。结论抑制Follistatin表达可促进人类BMSC细胞活性和BMP-2诱导人类BMSC成骨分化能力,Follistatin可抑制人类BMSC成骨分化。

Objective To investigate the influence of Follistatin depression on bone morphogenetic protein （BMP）‐2‐induced osteogenic differentiation of human bone marrow derived mesenchymal stem cells （BMSCs） and cell viability .Methods Follistatin‐specific small interfering RNA （siRNA） was transfected into human BMSCs ,the mitochondrial dehydrogenase activity ,protein amount and cellular DNA content were measured . The expression levels of some osteoblastic genes were analysed by quantificational reverse transcriptase polymerase chain reaction （qRT‐PCR） ,the influence of Follistatin depression on BMP‐2‐induced osteogenic differentiation of human BMSCs was assessed by alkaline phosphatase （ALP） staining and alizarin red staining .Results Our research indicated that DNA content and human BMSCs metabolism increased remarkably after 3 days and 7 days ,and total protein amount increased after 14 days .The expression levels of ALP ,OCN ,OSX ,OC ,OPN and RUNX2 increased ,ALP activity and calcium deposits increased after Follistatin depression .Conclusion Human BMSC viability and BMP‐2‐induced osteogenic differentiation of human BMSC increases after Follistatin depression , which shows that Follistatin is inhibit factor on osteogenic differentiation of human BMSCs .