人重组肿瘤坏死因子-α体外诱导人小梁细胞凋亡

Human recombinant tumor necrosis factor-alpha induced apoptosis in human trabecular meshwork cell

目的：研究人重组肿瘤坏死因子-α（rhTNF—α）对人小梁细胞（human trabecular meshwork cells，HTM）凋亡的影响，探讨其在原发性青光眼发病中的作用机制。方法：组织块法原代培养人小梁细胞，取第3～5代细胞用于实验。不同浓度的rhTNF—α与细胞共同孵育24h，采用Annexin—V联合PI双染流式细胞术，测定细胞凋亡率，结合荧光显微镜来观查凋亡细胞。结果：浓度低于0．01MU／L的肿瘤坏死因子-α作用24h对人小梁细胞凋亡无明显影响，浓度0．01MU／L及以上的肿瘤坏死因子-α作用24h可明显增加入小梁细胞凋亡率，凋亡率与肿瘤坏死因子-α浓度成正比。结论：肿瘤坏死因子-α可以诱导体外培养的人小梁细胞发生凋亡，可能参与了青光眼发病中小梁细胞的损害过程。

AIM： To investigate the effects of the human recombinant tumor necrosis factor- alpha （rhTNF-α） on the apoptosis of human trabecular meshwork （HTM） cell. METHODS： Primary and passage culture of HTM cells were established in vitro. The 3-5 passages of HTM cells were incubated with different concentrations of rhTNF-α for 24 hours, the apoptosis of HTM cells were detected by the method of Annexin V-FITC and PI stain, and the apoptosis rates were determined by the flow cytometer. The fluorescence microscope was used to recorded the apoptosis cells, which were stained by Annexin V-FITC. RESULTS： rhTNF-α at the concentration 0.01MU/L and above induced apoptosis of the HTM cells, the apoptosis rate were raised depending on the concentration; while under 0.01MU/L, rhTNF-α seems has no effects on the HTM cells. CONCLUSION： rhTNF-α can induce apoptosis of the cultured HTM cells in vitro, rhTNF-α may play an important role in the progress of glaucoma by damaging the HTM cells.