丙戊酸钠通过p21调控大鼠神经干细胞的增殖  

Sodium valproate inhibits proliferation in rat neural stem cells through p21 pathway

作　　者：黄磊[1] 储卫华[1] 袁继超[1] 赵明月[1] 陈图南[1] 蒋周阳[1] 林江凯[1] 冯华[1] 

机构地区：[1]第三军医大学西南医院神经外科,全军神经外科研究所,全军神经创伤防治重点实验室,重庆400038

出　　处：《第三军医大学学报》2013年第6期487-490,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(81000531);第三军医大学青年创新基金(2009XQN27)~~

摘　　要：目的探讨丙戊酸钠(sodium valproate,VPA)对体外培养的成年雌大鼠脊髓神经干细胞(neural stem cells,NSCs)增殖的影响。方法采用不同浓度的VPA(10-5、10-4、10-3、10-2、10-1、1、10 mmol/L)作用于NSCs,CCK-8法检测在不同时间点(0、24、48、72 h)对细胞增殖的影响;VPA(10-5、1 mmol/L)作用于NSCs 48 h后,流式细胞仪测定细胞周期分布,PCR测定p21在基因水平的表达,Western blot测定p21在蛋白质水平的表达。结果 CCK-8检测显示,当VPA浓度>10-5mmol/L时,体外培养的成年大鼠脊髓NSCs的增殖受到明显抑制,且具有时间依赖性。流式细胞仪细胞周期检测显示,同样浓度下,VPA可阻滞NSCs由G0/G1期向S期转换,表现为G0/G1期细胞增多,S期细胞减少,G2/M期细胞减少。PCR检测发现VPA可促进p21基因水平的表达。Western blot检测发现VPA可促进p21蛋白质水平的表达,各组间差异均有统计学意义(P<0.05)。结论 VPA可能通过促进p21表达,使NSCs阻滞于G0/G1期,最终抑制NSCs增殖。Objective To determine the effect of sodium valproate （VPA） on the proliferation and cell cycle in adult female rat spinal neural stem cells （NSCs）. Methods Cell proliferation was assessed by CCK-8 assay after the cells were treated with VPA at different concentrations of 10-5, 10-4, 10-3, 10-2, 10 - l, 1 or 10 mmol./L for 0, 24, 48 or 72 h. After NSCs were treated with VPA at a dose of 10-5 and 1 mmol/L for 48 h, cell cycle was analyzed by flow cytometry and the expression of p21 （cyclin-dependent kinase inhibi- tor） was detected by PCR and Western blot analysis. Results CCK-8 staining colorimetry showed that the proliferation of NSCs was markedly inhibited in a time-dependent manner when the concentration of VPA was more than 10-5mmol/L. Flow cytometry indicated more cultured NSCs were arrested in the G0/G1 phase and fewer at the S phase after being treated with VPA, which indicated that VPA arrested the transition of NSCs from G0/G1 phase to S phase. PCR and Western blot analysis indicated that VPA enhanced the expression of p21 at mRNA and protein levels（P 〈0.05）. Conclusion VPA may arrest NSCs at G0/G1 by increasing the expression of p21, and then finally inhibit the proliferation of NSCs.

关 键 词：丙戊酸钠 脊髓 神经干细胞 增殖 P21 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]