| 目的 观察氯离子通道阻断剂5-硝基-2-(3-苯丙胺)苯甲酸(NPPB)对培养牛角膜内皮细胞增殖中的作用。 方法 实验设5组，对照组，0.1%DMSO组， 50 、100 、200 µmol/L NPPB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(OD)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100 µmol/L NPPB处理48 h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 study the effect of NPPB on cultured bovine corneal endothelium cells in vitro. Methods The experiments was assembled to five groups, including control group, 0.1%DMSO group， 50 、100 、200μmol/L NPPB groups respectively. MMT assay was used to observe cell proliferation and cell survival rate was calculated. The phase change of cell cycle was used to examine by flow cytometry(FACS) and cell proliferation index was calculated. Results Compared with control group, the MTTOD value, cell survival rate of cultured bovine corneal endothelium cells in vitro decreased significantly with different concentration of NPPB groups, and showed a dose-dependent correlation. After treated with 100μmol/L NPPB, the rate of G1 stage increased, the rate of S stage and G2/M stage and cell proliferation index meanwhile decreased significantly. Conclusion Chloride channel inhibitor NPPB had the inhibition effect on cultured bovine corneal endothelium cells, and resulted in the cell cycle was stagnant at G1 stage. |
