GDF-5对大鼠骨髓间充质干细胞增殖、迁移的影响及相关作用机制  

Effect of GDF-5 on proliferation and migration of rat bone marrow mesenchymal stem cells and its related mechanism

作　　者：胡广 关智宇 张开伟 HU Guang;GUAN Zhi-Yu;ZHANG Kai-Wei(Department of Orthopedics,The First Affiliated Hospital of Guiyang College of Traditional Chinese Medicine,Guiyang 550001,Guizhou,China)

机构地区：[1]贵州中医药大学第一附属医院骨科,贵州贵阳550001

出　　处：《中国老年学杂志》2022年第21期5289-5293,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金委员会资助项目(81960813)。

摘　　要：目的探究生长分化因子(GDF)-5对大鼠骨髓间充质干细胞(BMSCs)增殖、迁移的影响及相关作用机制。方法购买10只SPF级SD大鼠,分离BMSCs,培养后分为对照组、下调组、上调组,并通过细胞转染建立稳定转染的下调组(GDF-5下调)和上调组(GDF-5上调)。采用流式细胞仪检测BMSCs的细胞周期,采用噻唑蓝(MTT)法检测细胞增殖能力,采用流式细胞仪检测细胞凋亡能力和周期分布情况,采用Transwell小室检测细胞侵袭,采用细胞划痕实验检测细胞迁移,采用Western印迹检测各组细胞磷脂酸肌醇3激酶/蛋白激酶B(PI3K/AKT)信号通路蛋白PI3K、AKT表达量。结果与下调组相比,上调组GDF-5表达量明显升高,G1期明显降低,S期、G2期明显升高,不同时间点增殖率明显上升,凋亡率明显降低,细胞迁移明显上升,侵袭数明显下降,PI3K、AKT信号通路蛋白表达量明显降低(均P<0.05)。结论上调GDF-5作用于PI3K/Akt信号通路,促进PI3K、AKT表达,进而促进BMSCs凋亡,抑制BMSCs增殖、侵袭、迁移。Objective To explore the effect of growth differentiation factor(GDF)-5 on the proliferation and migration of rats bone marrow mesenchymal stem cells(BMSCs) and its related mechanism.Methods Ten SPF SD rats were purchased and BMSCs isolated were divided into control group, down regulation group and up regulation group after culture.Stable transfection down regulation group(down regulation of GDF-5) and up regulation group(up regulation of GDF-5) were established by cell transfection.The cell cycle of BMSCs was detected by flow cytometry.Thiazole blue(MTT) assay was used to detect cell proliferation, flow cytometry was used to detect the cell apoptotic ability and cycle distribution.Cell invasion was detected by Transwell chamber, cell migration was detected by cell scratch test, the expression of phosphatidylinositol 3 kinase/protein kinase B(PI3 K/AKT) signaling pathway protein PI3 K and AKT in each group were detected by Western blot.Results Compared with down regulation group, the expression of GDF-5 was significantly increased, the G1 phase was significantly decreased, the S phase and G2 phase were significantly increased, the proliferation rate was significantly increased at different time points and the apoptosis rate was significantly decreased, the cell migration was significantly increased and the invasion number was significantly decreased, the protein expression of PI3 K and AKT signaling pathway were significantly decreased in up regulation group(all P<0.05).Conclusions Up regulation of GDF-5 could promote the expression of PI3 K and AKT through PI3 K/AKT signaling pathway, thus promoting the apoptosis of BMSCs and inhibiting the proliferation, invasion and migration of BMSCs.

关 键 词：生长分化因子5 骨髓间充质干细胞 增殖 迁移 

分 类 号：R-332[医药卫生]