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目的观察氯离子通道阻断剂5-硝基-2-(3-苯丙胺)苯甲酸[5-nitro-2-(3-phenyl-propylamino)-benzoic acid,NPPB]对培养牛角膜内皮细胞增殖的作用。方法实验设5组,对照组,0.1%二甲基硫氧化物DMSO(di mathyl sulfoxide)组,50μmol·L-1、100μmol·L-1、200μmol·L-1NPPB组。应用MTT比色分析法观察细胞增殖情况,计算细胞存活率。采用流式细胞仪检测细胞周期时相的变化,计算细胞增殖指数。结果NPPB使培养牛角膜内皮细胞MTT光吸收值和细胞存活率较对照组均显著降低,并具有浓度和时间依赖性。经100μmol·L-1NPPB处理48h后,与对照组相比,细胞出现明显的凋亡峰,G1期细胞比例明显增高,S期及G2期细胞比例则明显降低,增殖指数明显下降。结论NPPB能浓度依赖性抑制培养牛角膜内皮细胞的增殖,并使细胞停滞于G1期。
Objective To observe the effects of 5-nitro-2- (3-phenyl-propylamino) -benzoic acid (NPPB), a chloride channel blocker, on the proliferation of cultured bovine corneal endothelial cells effect. Methods The experiment was divided into 5 groups: control group, 0.1% dimethyl sulfoxide (DMSO) group, 50μmol·L-1, 100μmol·L-1,200μmol·L-1NPPB group. MTT colorimetric assay was used to observe cell proliferation and cell viability was calculated. Flow cytometry was used to detect the changes of cell cycle phase and calculate the cell proliferation index. Results NPPB decreased the MTT light absorbance and cell viability of cultured bovine corneal endothelial cells more significantly than that of the control group in a concentration- and time-dependent manner. After treated with 100μmol·L-1NPPB for 48h, the cells showed obvious apoptotic peak compared with the control group, the proportion of cells in G1 phase increased significantly, the proportion of cells in S phase and G2 phase decreased significantly, and the proliferation index decreased significantly. Conclusion NPPB can inhibit the proliferation of cultured corneal endothelial cells in a concentration-dependent manner and arrest the cells in G1 phase.