细胞因子对培养的人视网膜色素上皮细胞早期生长反应基因-1的影响  

Effects of cytokines on early growth response gene-1 in cultured human retinal pigment epithelial cells

作　　者：王静波[1] 张延军[2] 惠延年[1] 

机构地区：[1]第四军医大学西京医院眼科,西安710033 [2]广州军区广州总医院药学部

出　　处：《中华眼底病杂志》2007年第6期410-413,共4页Chinese Journal of Ocular Fundus Diseases

基　　金：西安市科技攻关项目（GG05163）

摘　　要：目的检测细胞因子对人视网膜色素上皮（RPE）细胞早期生长反应基因-1（Egr1）的影响。方法体外培养人RPE细胞，采用免疫荧光染色、Western blotting和逆转录聚合酶链反应（RTPCR）定性定量观察不同刺激因素作用下Egr-1蛋白和mRNA的表达变化。刺激因素包括：20μg／ml脂多糖（LPS）、40ng／ml肿瘤坏死因子（TNF）-α、10U／ml干扰素（IFN）-γ、30％单核／巨噬细胞株（THP1细胞）七清液和正常人玻璃体液分别刺激O（未受刺激）、10、20、30、40、60min。结果在未受刺激的RPE细胞中Egr-1呈弱阳性表达，细胞浆为浅黄绿色荧光，随着刺激因素的作用，Egr-1的表达明显增强，细胞浆和部分细胞核呈强绿色荧光。20μg／mlLPS、40ng／mlTNF—α、10U／mlIFN-γ、30％THP-1细胞上清液和玻璃体液刺激RPE细胞后，与未受刺激的RPE细胞相比，Egr-1 mRNA的最大增强幅度分别为1．9、1．3、1．4、1．2、1．4倍，Egr1蛋白的最大升高幅度分别为3．4、1．2、1．7、3．2、1．3倍。结论LPS、TNF—α、IFN-7、THP-1细胞上清液和玻璃体液可上调体外培养的人RPE细胞Egr-1 mRNA和蛋白的表达，且出现核转位现象，说明这些刺激因素可引起Egr-1的活化。Objective To detect the effects of cytokines on the expression of early growth response gene-1 （Egr-1） in cultured human retinal pigment epithelial （RPE） cells. Methods Immunofluorescence staining, Western blotting and reverse transcription polymerase chain reaction （RT-PCR） were used to detect and quantitatively analyze the expression of Egr-1 protein and mRNA in cultured human RPE cells which were exposed to stimulants, including 20μg/ml lipopolysaccharide （LPS）, 40 ng/ml tumor necrosis factor （TNF）-α, 10 U/ml interferon （IFN）-γ, 30% supernatant of monocyte/macrophage strain （THP-1 cells ） and the vitreous humor from healthy human eyeballs, for 0, 10, 20, 30, 40 and 60 minutes, respectively. Results The RPE cells stimulated for 0 minute revealed faint green fluorescence of Egr-1 in the cytoplasm. With exposure to the stimulants, the expression of Egr 1 increased obviously and strong green fluorescence was found in cytoplasm in some nuclei of RPE cells. Compared with the untreated RPE cells, after stimulated by 20μg/ml LPS, 40 ng/ml TNF a, 10 U/ml IFN Y, 30% supernatant of THP-1 cells and the vitreous humor, the approximate ultimate amplitudes of Egr-1 mRNA enhanced 1.9, 1.3, 1.4, 1.2, and 1.4 times, respectively ; the greatest amplitudes of Egr 1 protein increased 3.4, 1.2, 1.7, 3.2, and 1. 3 times, respectively. Conclusion LPS, TNF-α, IFN-γ, supernatant of THP 1 cells and the vitreous humor can up-regulate the expression of Egr-1 mRNA and protein in cultured human RPE cells, and induce its nuclear transposition, which suggests the activation of Egr-1.

关 键 词：色素上皮 眼 肿瘤坏死因子Α 脂多糖类 干扰素Γ 重组 早期生长反应蛋白质1 

分 类 号：R686[医药卫生—骨科学]