中华骨质疏松和骨矿盐疾病杂志Issue(3)：250-257,8.DOI:10.3969/j.issn.1674-2591.2014.03.011
PI3 K／Akt信号通路在骨髓间充质干细胞增殖及成骨分化调控中的作用
Roles of PI3K/Akt signaling pathway in regulating bone mesenchymal stem cells proliferation and differentiation
摘要
Abstract
Objective To study the effect LY294002, a PI3K/Akt signaling pathway inhibitor, on the prolifer-ation and differentiation of human bone mesenchymal stem cells .Methods Human mesenchymal stem cells ( hMSCs ) were isolated by adherent culture and identified .hMSCs were treated with LY294002 , the PI3K specific inhibitor , at two different concentrations , 1 μmol/L or 10 μmol/L, and the cell proliferation was tested by MTT assay .After 3 days or 7 days osteogenic induction culture , osteogenic differentiation was observed by ALP staining and qualification .Alizarin red staining and determination was adopted to analyze mineralization degree .Western blot analysis was employed for Akt phosphorylation .The gene expressions of osteogenic markers , such as BMP2, Runx2, osteopontin and Osterix , were de-termined by Realtime-PCR.Results In non-induction conditions from 24 hours to 72 hours, LY294002 significantly suppressed hMSCs proliferation and the suppressive effect was increased as time went by ( P<0.05 ) .10μmol/L showed the highest ALP activity over the control and 1μmol/L groups in ALP staining and quantitation at 3 days or 7 days osteo-genic induction (P<0.05).Similarly, 1 μmol/L and 10 μmol/L were more extensively calcified than the control in Alizarin red staining and determination after 3 days or 7 days osteogenic induction (P<0.05).At 7 days, 10 μmol/L contained more calcified minerals that 1μmol/L ( P<0.05 ) .Western blot analysis revealed that osteogenic induction stimulated the phosphorylation of Akt which could be inhibited by LY 294002 . At 7 days osteogenic culture , either 1 μmol/L or 10 μmol/L increased the mRNA expression levels of BMP2, Runx2, osteopontin and Osterix (P<0.05). Conclusion PI3K/Akt signaling pathway is involved with the process of proliferation and differentiation in hMSCs .Dur-ing osteogenic differentiation , activation of PI3K/Akt differentially regulates downstream effectors Akt .LY294002 arrests hMSCs proliferation while promots their osteogenic differentiation and subsequent mineralization .关键词
PI3K/Akt信号通路/骨髓间充质干细胞/细胞增殖/成骨分化Key words
PI3K/Akt signaling pathway/mesenchymal stem cells/cells proliferation/osteogenic differentiation分类
医药卫生引用本文复制引用
王雪鹏,李茂强,边振宇,季成,何齐芳,姚旺祥,朱六龙..PI3 K／Akt信号通路在骨髓间充质干细胞增殖及成骨分化调控中的作用[J].中华骨质疏松和骨矿盐疾病杂志,2014,(3):250-257,8.基金项目
卫生部医药卫生科技发展中心医学科研专项课题（ W2013 ZT205）；浙江省卫生厅一般项目（02012KYA146）；杭州市卫生局重点项目 （）
