炎性细胞因子对色素上皮细胞衍生因子表达的影响

The influence of inflammatory cytokines to the expression of pigment epithelium-derived factor

目的观察肿瘤坏死因子（TNF—α）、白细胞介素（IL）-6、IL-8等炎性细胞因子对视网膜色素上皮细胞衍生因子（PEDF）表达的影响。方法原代培养人视网膜色素上皮（RPE）细胞，取3～5代细胞分别以浓度为20．00、2．00、0．20、0．02ng／ml的TNF—α、IL-6、IL-8干预6、12、24、48h，蛋白免疫印迹方法检测培养液中PEDF蛋白质的表达水平。结果TNF—α、IL-6、IL-8对人RPE细胞PEDF的分泌均有抑制作用，并且其抑制作用随着3种炎性细胞因子浓度的增加和作用时间的延长而增强；在浓度0．02ng／ml、作用时间6h（F=7．14，P〈0．05），浓度2．00ng／ml、作用时间48h（F=14．05，P〈0．01），浓度20．00ng／ml、作用时间24h（F=11．53，P〈0．01）3种作用条件下，对PEDF分泌抑制作用的差异具有统计学意义，其中TNF—α干预组PEDF的表达量最低，对人RPE细胞PEDF的分泌抑制作用最明显（F=14，P〈0.01）。结论TNF-α、IL-6、IL-8能够下调体外培养的人RPE细胞PEDF蛋白质的表达。

Objective To evaluate the effects of inflammatory cytokines, including tumor necrosis factorTNF-α and interleukins （IL 6 and IL-8）, to the expression of pigment epithelium derived factor （PEDF） in human retinal pigment epithelium （RPE）cells. Method Cultured primary human RPE cells were treated with 20,2,0.2, and 0.02 ng/ml of TNF-α, IL-6 and IL-8 separately. The levels of PEDF expression were determined by Western blot of the supernant after 6,12,24 and 48 hours of culture. Results PEDF secretion of RPE cells was inhibited by TNF-α, IL-6 and IL-8 in a time and dose-dependent fashion. Compared with the controls, the expression of PEDF decreased significantly in 0. 02 ng/ml and 6 hours group （F= 7.14, P〈0. 05）, 2.00 ng/ml and 48 hours group（F= 14.05,P〈0.01） , and 20. 00 ng/mt and 24 hours group（F=11.53,P〈0.01）. TNF -αwas the most strength inhibitor （F= 14,P〈0.01）. Conclusion TNF-α, IL-6, and IL-8 could suppress the expression of PEDF in the cultured human RPE cells.