论文部分内容阅读
目的 探讨在不同缺氧时相人视网膜色素上皮(RPE)细胞转录和表达缺氧诱导因子-1α(HIF -1α)的情况。方法 原代培养人RPE细胞,经鉴定后,选择第3 ~6代RPE细胞进行缺氧分析,分别在缺氧6、8、16、24、48h不同时相,利用半定量RT PCR法及免疫荧光测定法检测人RPE细胞转录和表达HIF -1α的情况。结果 半定量RT- PCR法和免疫荧光法检测结果均显示缺氧可以明显诱导人RPE细胞转录和表达HIF- 1α。在缺氧8h时,HIF- 1α转录和表达量达高峰,并可持续至24h后;缺氧48h时,人RPE细胞代谢产物增加,细胞形态发生改变,HIF 1α转录和表达量开始下降。结论 缺氧可以明显诱导人RPE细胞转录和表达HIF- 1α,这可能是发生脉络膜新生血管的重要始动因素。
Objective To investigate the transcription and expression of hypoxia inducible factor-1α (HIF-1α) in human retinal pigment epithelium (RPE) cells under different hypoxia. Methods Human RPE cells were cultured in primary culture. After identified, RPE cells of passage 3 to 6 were selected for hypoxia analysis. The cells were cultured in different phases of hypoxia for 6, 8, 16, 24 and 48 hours respectively. Fluorescence assay was used to detect the transcription and expression of HIF-1α in human RPE cells. Results Both semi-quantitative RT-PCR and immunofluorescence assay showed that hypoxia induced the transcription and expression of HIF-1α in human RPE cells. After hypoxia for 8h, the transcription and expression of HIF-1α peaked and lasted for 24 h. After hypoxia for 48 h, the metabolites of human RPE cells increased, the morphology of cells changed and the transcription and expression of HIF-1α began to decrease. Conclusion Hypoxia can obviously induce the transcription and expression of HIF-1α in human RPE cells, which may be an important initiating factor for choroidal neovascularization.