小檗碱通过激活PI3K/AKT通路促进大鼠骨髓间充质干细胞的体外迁移

Berberine promotes the in vitro migration of bone marrow mesenchymal stem cells via activation of PI3K/AKT pathway

背景:小檗碱是中药黄连的主要生物碱,对骨髓间充质干细胞有多种活性作用。目的:观察小檗碱对大鼠骨髓间充质干细胞体外迁移的影响以及其作用机制。方法:采用全骨髓法分离培养SD大鼠骨髓间充质干细胞,调整细胞浓度至1.5×10~9 L^(-1)备用。小檗碱用二甲基亚砜溶解,并用含体积分数为1%胎牛血清的α-MEM培养基稀释,分别将浓度调整为1,3,10μmol/L。MTT法检测小檗碱对骨髓间充质干细胞活性的影响,Transwell小室和划痕实验检测小檗碱对大鼠骨髓间充质干细胞迁移的影响,Western blot检测小檗碱对p-AKT通路调控作用。结果与结论:(1)小檗碱对骨髓间充质干细胞的活力无影响;(2)小檗碱在10μmol/L浓度下能够显著促进骨髓间充质干细胞的迁移,划痕实验结果与Transwell实验相一致;(3)加入小檗碱可以促进骨髓间充质干细胞中p-AKT表达,加入p-AKT抑制剂LY294002能够逆转小檗碱的促迁移作用;(4)结果表明,小檗碱能够通过激活PI3K/AKT通路促进大鼠骨髓间充质干细胞的迁移。

BACKGROUND:Berberine as the main alkaloid of herbal Coptis chinensis has many pharmacological effects on bone marrow mesenchymal stem cells(BMSCs).OBJECTIVE:To explore the effects and mechanism of berberine on the in vitro migration of BMSCs.METHODS:Rat BMSCs were isolated and cultured by a whole bone marrow assay,and prepared into 1.5×109/L.Berberine was dissolved by dimethyl sulfoxide,and diluted byα-MEM containing 1%fetal bovine serum into the concentrations of 1,3,10μmol/L.Cell migration was assessed using a Transwell chamber and cell scratch assay.Western blot assay was used to test the regulatory effect of berberine on p-AKT pathway.RESULTS AND CONCLUSION:Berberine had no effect on the cell viability of BMSCs.Berberine at 10μmol/L significantly promoted the migration of BMSCs.The cell scratch assay shared consistent findings with the Transwell assay.Addition of berberine up-regulated the p-AKT expression in BMSCs,while LY294002,the inhibition of p-AKT,could effectively reverse the pro-migration effects of berberine.Overall,berberine can promote the migration of BMSCs via the activation of PI3K/AKT pathway.