缺氧诱导人视网膜色素上皮细胞整合素连接激酶的表达及其与HIF-1α、VEGF的相关性

Hypoxia-induced ILK expression in human retinal pigment epithelial cells and its association with the expressions of HIF-1α and VEGF

目的:探讨缺氧条件下人视网膜色素上皮细胞(hRPE)整合素连接激酶(integrim linked kinase,ILK)的表达,及其与缺氧诱导因1α(hypoxia induciable factor,HIF-1α)及血管内皮生长因子(vascular cndothclial growth factor,VEGF)表达的相关性。方法:hRPE培养,COCL2(150"mol/L)建立化学缺氧细胞模型。免疫组化方法检测缺氧0、6、12、24、48、72hhRPE细胞ILK、HIF-1α、VEGF的表达。Western blot和RT-PCR方法检测以上缺氧时间细胞ILK、HIF-1α、VEGF的蛋白及其mRNA表达水平并进行图像分析。结果:ILK表达于正常培养的hRPE中,随着缺氧时间的延长,其表达增加,且与HIF-1α、VEGF的表达呈正相关性。结论:ILK可能作为低氧反应性因子与HIF-1α、VEGF共同参与hRPE的低氧病理过程。

Objective To study the expression of integrim linked kinase （ILK）in human retinal pigment epithelial （hRPE） cells under hypoxic conditions and its association with the expressions of hypoxia induciable factor-1α （HIF-1α） and vascular endothelial growth factor（VEGF）. Methods hRPE cells were cultured and a model of chemical hypoxia was established by COCl2 of 150 μmol/L. Immunohistochemical assay and Western blot were applied to detect the expression levels of ILK, HIF-1α, and VEGF proteins at different time points （0, 6, 12, 24, 48, and 72 h） during hypoxia while RT-PCR was used to detect mRNA expression levels of ILK, HIF-1α, and VEGF. The images were analyzed by the image analysis system. Result ILK was expressed in hRPE cells, with an increase in the expression level with prolonged hypoxia, and positively associated with the expressions of HIF-1α and VEGF. Conclusion ILK together with HIF-1α and VEGF may be involved in the pathologic process of hypoxia in human retinal pigment epithelial cells.