DCPIB对小胶质细胞株BV-2细胞增殖的抑制作用  

Inhibitory effect of DCPIB on proliferation of mouse microglia cell line BV-2

作　　者：李正伟[1] 陈劲草[1] 颜青[1] 张华楸[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经外科,武汉430030

出　　处：《中华实验外科杂志》2012年第3期456-458,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30900449）;教育部新教师基金资助项目（20090142120011）

摘　　要：目的探讨容积调控性氯离子通道（VRACs）阻断剂DCPIB对小胶质细胞BV-2细胞增殖的影响及作用机制。方法不同浓度的DCPIB（10、20、40μmol／L）作用于BV-2细胞相应的时间后，用噻唑蓝（Mar）比色法检测DCPIB对细胞增殖的抑制率；流式细胞术检测药物处理后BV-2细胞周期的分布；Westernblot检测细胞周期相关蛋白CyclinD1的表达。结果VRACs阻断剂DCPIB可使细胞生长曲线下移，呈时间和浓度依赖性；DCPIB抑制细胞增殖，作用细胞48h后的抑制率分别为（29．20±3．99）％、（38．93±2．36）％、（68．52±2．16）％，与正常对照组比较均有明显增高（P〈0．01）。细胞周期阻滞在Go／G，期，G2／M期和S期细胞明显减少。DCPIB作用BV-2细胞72h后，与对照组比较，CyclinD1蛋白表达明显降低。结论VRACs阻断剂DCPIB可以抑制BV-2细胞增殖，其作用机制可能与阻滞细胞周期和下调CyclinD1蛋白的表达有关。Objective To siudy the inhibitory effect of DCPIB on mouse microglia cell line BV-2 and the mechanisms underlying such effect. Methods The microglia cell line BV-2 was incubated with DCPIB in different concentrations （10, 20 and 40 trmol/L） for different intervals （24, 48 and 72 h）. Cell inhibition ratio was assessed by methyl thiazol tetrazolium （MTY） assay and flow cytometry analysis was performed to detect the cell cycle. Furthermore, Cyclin D1 expression was detected by using Western blotting. Results MTY assay revealed that BV-2 cell growth curve moved down in a concentration- and time- dependent manner. Cell grown was inhibited by DCPIB obviously, and the inhibition ratio was （29. 20 ± 3.99）%, （38.93 ±2.36）%, （68.52 ±2. 16）% at 48 h after treatment with DCPIB （10, 20 and 40 μmol/L）. Flow cytometry showed that most BV-2 cells were inhibited by DCPIB and arrested in G0/G, phase, while proportion of BV-2 cells in G2/M phase and S phase was decreased. Western blotting demon- strated that the expression level of Cyclin D1 was down-regulated in BV-2 ceils after treatment with DCPIB for 72 h. Conclusion The results suggest that DCPIB can markedly inhibit the proliferation of BV-2 cells, which may be related with arrest of the cell cycle in G0/G1 phase and down-regulation of Cyclin D1 expression.

关 键 词：小胶质细胞 氯离子通道阻断剂 增殖 细胞周期 

分 类 号：R363[医药卫生—病理学]