氯离子通道阻滞剂对人眼小梁细胞吞噬作用的影响

Effect of chloride channel inhibitor in phagocytic process of human trabecular meshwork cells

目的探讨氯离子通道阻滞剂—5-硝基-2-(3-苯丙胺)苯甲酸5-nitro-2-(3-phenyl-propylamino)-benzoic acid,NPPB对人眼小梁细胞吞噬功能的影响。方法培养人眼小梁细胞,对其进行免疫化学染色鉴定,采用RT-PCR技术检测小梁细胞CLC-2mRNA的表达,然后应用流式细胞仪定量检测不同浓度NPPB作用下,小梁细胞对乳胶株的吞噬指数。结果体外培养的人眼小梁细胞NES染色阳性,且成功检测出CLC-2mRNA表达。当加入10μmol/L的NPPB作用6h后,小梁细胞对乳胶株的吞噬指数与对照组相比无明显改变(P>0.05),而50μmol/L的NPPB作用后,小梁细胞对乳胶株的吞噬作用明显降低(P<0.01),低于对照组,且随着加入NPPB浓度的增加,吞噬指数逐渐降低。结论氯离子通道阻滞剂浓度依赖的抑制小梁细胞的吞噬功能,且氯离子通道CLC-2可能参与调节小梁细胞吞噬过程。

Objective To investigate the effect of chloride channel inhibitor 5-nitro-2-（3-phenyl-propylamino）-benzoic acid（NPPB） on cultured human trabecular meshwork cells phagocytic process. Methods Cultured human trabecular meshwork cells was assessed by immunochemistry and the expression of CLC-2 mRNA in cultured human trabecular meshwork cells was detected with RT-PCR. Then, flow cytometry was performed to detect the phagocytic index on cultured human trabecular meshwork cells which had been pre-treated by chloride channel inhibitor NPPB. Results Immunohistochemical staining of the cultured human trabecular meshwork cells which was detected the expression of CLC-2 mRNA was positive. Compared with that of control group, 10 μmol/L NPPB didn′t affect the phagocytic process of the trabecular meshwork cells（P〈0.05）, however when treated with 50, 100 and 500 μmol/L DNDS, the phagocytic indexes were significantly decreased（P〈0.01）,and as the concentration were increased, the phagocytic indexes was decreased gradually. Conclusions The trabecular meshwork cell′s phagocytic process is affected by a certain concentration of NPPB and chloride channel CLC-2 may be play important roles in the phagocytic process of trabecular meshwork cells.