中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (19): 3453-3456.doi: 10.3969/j.issn.1673-8225.2012.19.007

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

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★

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   

  1. 徐州医学院附属医院,1急救中心,2中心实验室,   3妇产科,江苏省徐州市  221002
  • 收稿日期:2011-11-30 修回日期:2012-01-16 出版日期:2012-05-06 发布日期:2012-05-06
  • 通讯作者: 许铁,硕士生导师,教授,主任医师,徐州医学院附属医院急救中心,江苏省徐州市221002 xutie889@163.com
  • 作者简介:刘筱★,男,1983年生,江苏省丰县人,汉族,徐州医学院在读硕士,医师,主要从事骨髓间充质干细胞及脊髓损伤修复的研究。liuxiao628@hotmail.com

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

Liu Xiao1, Wan Mei-rong2, Liu Xiao-yun2, Yan Xian-liang1, Zhang Zheng-zheng3, Liu Kai1, Xu Tie1   

  1. 1Emergency Care Center, 2Central Laboratory, 3Department of Obstetrics and Gynecology, the Affiliated Hospital of Xuzhou Medical College, Xuzhou  221002, Jiangsu Province, China
  • Received:2011-11-30 Revised:2012-01-16 Online:2012-05-06 Published:2012-05-06
  • Contact: Xu Tie, Master’s supervisor, Professor, Chief physician, Emergency Care Center, the Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China xutie889@163.com
  • About author:Liu Xiao★, Studying for master’s degree, Physician, Emergency Care Center, the Affiliated Hospital of Xuzhou Medical College, Xuzhou 221002, Jiangsu Province, China liuxiao628@hotmail.com

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10 µmol/L Rho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,3 d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。
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
doi:10.3969/j.issn.1673-8225.2012.19.007

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

Abstract:

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.

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