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  

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

作　　者：朱余俊 吕浩 江渟 ZHU Yujun;LYU Hao;JIANG Ting(Graduate School,Anhui University of Chinese Medicine,Hefei 230031,China;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学研究生院,安徽合肥230031 [2]安徽中医药大学第一附属医院,安徽合肥230031

出　　处：《中国中医药信息杂志》2022年第3期79-84,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：安徽省重点研究与开发计划(202104j07020010);安徽中医药大学第一附属医院临床科学研究项目(2020yfyzc27)。

摘　　要：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%药物血清组,每组6个复孔,分别加入相应血清培养,CCK-8法检测细胞增殖活性,Western blot检测细胞PI3K、AKT、p-AKT蛋白表达,qPCR检测细胞PI3K、AKT、LRP5、β-catenin mRNA表达。结果不同浓度含药血清可促进HBMSCs增殖,且存在一定的量效和时效关系,尤以5%药物血清作用36 h促进细胞增殖作用最明显。与同浓度空白血清组比较,2%、5%药物血清组HBMSCs PI3K、AKT、p-AKT蛋白表达及PI3K、AKT、LRP5、β-catenin mRNA表达明显升高(P<0.05,P<0.01);与2%药物血清组比较,5%药物血清组HBMSCs PI3K、AKT、p-AKT蛋白表达及PI3K、AKT和β-catenin mRNA表达明显升高(P<0.05,P<0.01)。结论复方补肾活血颗粒可能通过促进HBMSCs增殖,上调PI3K、AKT、p-AKT蛋白及PI3K、AKT、LRP5、β-catenin mRNA表达,激活Wnt/PI3K-AKT信号通路,发挥治疗OP的作用。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.

关 键 词：骨质疏松症 骨髓间充质干细胞 复方补肾活血颗粒 细胞增殖 Wnt/PI3K-AKT信号通路 

分 类 号：R285.5[医药卫生—中药学]