Rho激酶抑制剂Y-27632干预大鼠骨髓间充质干细胞增殖及细胞周期的变化  

Effects of Rho kinase inhibitor Y-27632 on proliferation and cell cycle of rat bone marrow mesenchymal stem cells

作　　者：刘筱[1] 万美荣[2] 刘小云[2] 燕宪亮[1] 张铮铮[3] 刘凯[1] 许铁[1] 

机构地区：[1]徐州医学院附属医院急救中心,江苏省徐州市221002 [2]徐州医学院附属医院中心实验室,江苏省徐州市221002 [3]徐州医学院附属医院妇产科,江苏省徐州市221002

出　　处：《中国组织工程研究》2012年第19期3453-3456,共4页Chinese Journal of Tissue Engineering Research

摘　　要：背景:Rho激酶抑制剂可以调节细胞骨架重构,激活转录因子,促进细胞增殖和分化。目的:观察Rho激酶抑制剂Y-27632对大鼠骨髓间充质干细胞分裂增殖和细胞周期的影响。方法:采用贴壁细胞培养法体外分离培养SD大鼠骨髓间充质干细胞,并传代。取生长状态良好的第2代骨髓间充质干细胞分组:实验组加入10μmol/LRho激酶抑制剂Y-27632,对照组正常培养。结果与结论:①骨髓间充质干细胞增殖:细胞生长曲线呈"S"型,Rho激酶抑制剂Y-27632作用后,3d左右进入平台期,细胞生长加速,生长曲线上移。②骨髓间充质干细胞的细胞周期:对照组G0/G1期细胞比例为(80.640±5.516)%,S期细胞比例为(13.397±2.511)%;实验组G0/G1期细胞比例为(61.723±8.829)%,S期细胞比例为(29.380±7.630)%,实验组S期细胞比例高于对照组(P<0.05)。表明Rho激酶抑制剂Y-27632可以促进骨髓间充质干细胞DNA的合成,促进细胞分裂和增殖。BACKGROUND： Rho kinase inhibition can regulate dynamic reorganization of cytoskeletal proteins, activate transcriptional factor, and stimulate cell proliferation and differentiation. OBJECTIVE： To explore the effects of Rho kinase inhibitor Y-27632 on proliferation and cell cycle of bone marrow mesenchymal stem cells （BMSCs）. METHODS： Rat BMSCs were isolated using whole bone marrow culture method and purified using adherent culture method in vitro. After two passages, BMSCs were incubated in maintenance medium containing Y-27632 （10 μmol/L, a commonly used working concentration） in 96-well plates. A control group in which BMSCs were not treated was designated. RESULTS AND CONCLUSION： The growth curves of BMSCs for moderate inoculation density was typically S-shaped. After BMSCs were incubated with Rho kinase inhibitor Y-27632,the lag phase was found during the first 3 days, the growth was accelerated, and the growth curve was upward. Flow cytometry indicated that （80.640±5.516）% of the BMSCs were in G0/G1 phase of cell cycle, （13.397±2.511）% in S phase. After stimulated by Y27632, （61.723±8.829）% of the BMSCs were in G0/G1 phase, and （29.380±7.630） % were in S phase. The cell proliferation during S stage was significantly higher after stimulation by Y27632 （P 〈 0.05）. Rho kinase inhibitor Y-27632 can significantly promote more BMSCs to enter the stage of DNA synthesis, and promote the division and proliferation of BMSCs.

关 键 词：细胞周期 增殖 骨髓间充质干细胞 RHO激酶抑制剂 分离培养 大鼠 

分 类 号：R394.2[医药卫生—医学遗传学]