复方补肾活血颗粒对人骨髓间充质干细胞增殖及Wnt/PI3K-AKT信号通路相关蛋白表达的影响

目的 观察复方补肾活血颗粒药物血清对人骨髓间充质干细胞(HBMSCs)增殖的影响,探讨其干预骨质疏松症(OP)的作用机制.方法 从OP患者骨髓中分离培养HBMSCs,流式细胞仪检测HBMSCs表型和细胞周期;将HBMSCs分为正常组、2％空白血清组、2％药物血清组、5％空白血清组、5％药物血清组、8％空白血清组和8％药...

Effects of Compound Bushen Huoxue Granules on Proliferation of Bone Marrow Mesenchymal Stem Cells and Expression of Related Proteins in Wnt/PI3K-AKT Signaling Pathway

Objective To observe the effects of Compound Bushen Huoxue Granules drug-containing serum on the proliferation of human bone marrow mesenchymal stem cells(HBMSCs);To explore its mechanism of intervening in osteoporosis.Methods HBMSCs were isolated and cultured from the bone marrow of OP patients,and the phenotype and cell cycle of HBMSCs were detected by flow cytometry;HBMSCs were divided into normal group,2% blank serum group,2% drug-containing serum group,5%blank serum group,5% drug-containing serum group,8% blank serum group and 8% drug-containing serum group.Each group has 6 multiple wells and the corresponding serum was added for culturing.CCK-8 method was used to detect cell proliferation activity.Western blot was used to detect PI3K,AKT,p-AKT protein expressions.qPCR was used to detect PI3K,AKT,LRP5,β-catenin mRNA expressions.Results Different concentrations of drug-containing serum could promote the proliferation of HBMSCs,and there was a certain dose-effect and time-effect relationship.Especially,5%drug-containing serum had the most obvious effect on promoting cell proliferation after 36 hours.Compared with the blank serum group at the same concentration,the expressions of PI3K,AKT,p-AKT protein and PI3K,AKT,LRP5,and β-catenin mRNA of HBMSCs in 2% and 5% drug-containing serum groups significantly increased(P<0.05,P<0.01).Compared with the 2%drug-containing serum group,the expressions of PI3K,AKT,p-AKT protein and PI3K,AKT and β-catenin mRNA of HBMSCs in 5% drug-containing serum group significantly increased(P<0.05,P<0.01).Conclusion Compound Bushen Huoxue Granules may promote the proliferation of HBMSCs,up-regulate PI3K,AKT,p-AKT protein and PI3K,AKT,LRP5,β-catenin mRNA expressions,activate Wnt/PI3K-AKT signaling pathway,and then play a role in the treatment of osteoporosis.