缺氧诱导体外培养人视网膜色素上皮细胞中Toll样受体4的表达  

Expression of Toll-like recepter 4 in cultured human retinal pigment epithelium cells under hypoxic condition

作　　者：张雪[1] 王雨生[1] 叶子[1] 马可[1] 

机构地区：[1]第四军医大学西京医院眼科全军眼科研究所,中国陕西省西安市710032

出　　处：《国际眼科杂志》2010年第7期1260-1262,共3页International Eye Science

基　　金：中国国家自然科学基金资助项目(No.30371516;30672291)~~

摘　　要：目的:了解缺氧对人视网膜色素上皮(retinal pigmentepithelium,RPE)细胞Toll样受体4(Toll-like receptor4,TLR4)表达的影响。方法:采用200μmol/L CoCl2处理RPE细胞建立化学缺氧模型,以未处理细胞作对照,在缺氧处理后2,4,8,12和24h用免疫荧光法观察TLR4在细胞中的定位,用RT-PCR和Western blot检测细胞中TLR4表达水平。结果:共聚焦显微镜观察到正常对照组RPE细胞胞质内有微弱的TLR4荧光表达,缺氧后胞质内荧光表达明显增强,缺氧12h荧光强度达最强。RT-PCR和Western blot结果分别提示随缺氧时间延长,RPE细胞中TLR4mRNA和蛋白表达增强水平升高,缺氧至12h表达高峰最强(P<0.05),24h表达开始下降,各组差异有统计学意义。结论:缺氧可诱导RPE细胞中TLR4表达增强。AIM： To observe the effects of hypoxia on the expression of Toll-like receptor 4 （TLR4） in cultured human retinal pigment epithelium （RPE） cells. METHODS ： Chemical hypoxic model was established by treating the cultured human RPE cells with 200μmol/L CoCI2. The location of TLR4 in human RPE cells was observed by immunofluorescence assay, and RT-PCR together with Western blot were applied to detect the expression of TLR4 2, 4, 8, 12, 24 hours after the treatment. RESULTS： Immunofluorescence assay revealed that TLR4 was slightly expressed in human RPE cells under normoxic condition. After hypoxic treatment, there was a significant increase in green fluorescence within the cytoplasm. TLR4 expression tend to intensify over time and peaked at 12 hours, which was confirmed by RT-PCR and Western blot assay exhibiting TLR4 mRNA/protein levels （ P 〈 0.05）. The expression levels began to decrease at 24 hours, and there was statistical significance among the groups. CONCLUSION ： Hypoxia can strengthen TLR4 expression in cultured human RPE cells.

关 键 词：TOLL样受体4 缺氧 视网膜色素上皮细胞 

分 类 号：R774.1[医药卫生—眼科]