丹酚酸B通过激活PI3K/AKT通路促进人牙龈间充质干细胞增殖、迁移和成骨分化

Salvianolic acid B promotes the proliferation,migration and osteogenic differentiationo f human gingival mesenchymal stem cells by activating the PI3K/AKT pathway

目的探讨丹酚酸B(SalB)对人牙龈间充质干细胞(hGMSC)增殖、迁移和分化的影响。方法分离培养hGMSC,流式细胞术检测CD73、CD90、CD34、CD45表达;选取对数生长期细胞,使用(0、5、10)μmol/L SalB处理hGMSC 24 h。CCK-8法检测各组细胞的增殖活性,Transwell^(TM)实验和划痕实验检测细胞的迁移能力,碱性磷酸酶染色检测成骨能力、油红O染色检测成脂分化能力;实时定量PCR和Western blot法检测细胞相关分化蛋白和基因以及磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)信号通路相关分子的mRNA和蛋白表达。结果与空白组hGMSC比,SalB处理的hGMSC增殖、迁移能力显著增加,且呈剂量依赖性;hGMSC成骨能力和成骨相关蛋白、PI3K/AKT信号通路相关蛋白表达上调,成脂分化降低,成脂相关蛋白表达显著下调。结论SalB促进hGMSC的增殖、迁移,并促进hGMSC成骨分化、抑制成脂肪分化,可能与激活PI3K/AKT信号通路有关。

Objective To investigate the effect of salvianolic acid B(SalB)on the proliferation,migration and differentiation of human gingival mesenchymal stem cells(hGMSCs).Methods The hGMSCs were isolated and cultured,and the expressions of CD73,CD90,CD34,and CD45 were detected by flow cytometry;logarithmic phase cells were selected and hGMSCs were treated with 0,5,10μmol/L SalB for 24 hours.The proliferation activity of cells in each group was detected by CCK-8 assay,the migration ability of cells was detected by Transwell^(TM) assay and scratch test,and the osteogenic differentiation ability and the adipogenic differentiation ability were detected by alkaline phosphatase(ALP)staining and oil red O staining,respectively;the mRNA and protein expressions of cell differentiation proteins and genes as well as proteins related to the PI3K/AKT signal pathway were detected by Real-time quantiative PCR and Western blotting.Results The proliferation and migration ability of SalB-treated hGMSCs were significantly increased in a dose-dependent manner;the ability of osteogenic differentiation of hGMSCs and the expressions of osteogenesis associated proteins and PI3K/AKT signal pathway related proteins were up-regulated,while the adipogenic differentiation decreased,and the expression of adipogenesis related proteins was significantly down-regulated.Conclusion SalB promotes the proliferation,migration,and osteogenic differentiation of hGMSCs and inhibits the adipogenic differentiation,which may be related to the activation of PI3K/AKT signal pathway.