NPPB对星形胶质细胞增殖的抑制作用

The effect of NPPB on primary cultured astrocytes proliferation in vitro

目的观察体积调节性阴离子通道阻滞剂NPPB对体外培养星形胶质细胞增殖的影响。方法采用原代培养的大鼠大脑皮层星形胶质细胞,实验分为对照组(正常培养基培养)与NPPB干预组;在不同时间点(0、6、12、24、48h),应用流式细胞技术以及免疫细胞荧光双标法(Brdu/DAPI)检测各实验组星形胶质细胞增殖及细胞周期进展的情况。结果与对照组相比,NPPB干预组在12、24h时星形胶质细胞增殖率较正常对照组降低(P<0.05),同时处于S期细胞的百分率亦相对正常对照组明显下降(P<0.01)。结论体积调节性氯离子通道阻滞剂NPPB可以显著抑制体外培养星形胶质细胞增殖及细胞周期进展,提示VRAC通道参与了体外培养星形胶质细胞的增殖。

Objective To observe the effect of volume-regulated anion channel blocker（NPPB） on primary cultured astrocytes proliferation in vitro.Methods Primary cultured rat cortical astrocytes were randomly divided into two groups： control group and NPPB group（treated by DMEM/F12 containing 10% FBS with 100 μM NPPB or with 0.2% DMSO for control group）.At different time points（0 h,6 h,12 h,24 h and 48 h）,the changes of astrocytes proliferation and cell cycle progression were detected by flow cytometry and immunofluorescence staining（Brdu/DAPI）.Results Compared with the control group,the percentage of proliferation astrocytes was decreased at 12 h and 24 h after treatment（P0.05）.At the mean time,the penentage of astrocytes in Sphase in NPPB groups was significantly less（P0.01）,than control groups at these time points.Conclusions Volume-regulated anion channel blocker NPPB can remarkably inhibit astrocytes proliferation in vitro.Our results suggested that volume-regulated anion channel involved in normal cultured astrocytes proliferation.