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目的 观察人视网膜色素上皮 (retinal pigmentepithelium,RPE)细胞体外培养时的形态及相关细胞骨架蛋白变化规律 ,进而研究人玻璃体液对 RPE细胞形态及细胞骨架蛋白表达的影响 ,探讨玻璃体液在增生性玻璃体视网膜病变 (proliferative vitreoretinopathy,PVR)中的调控作用。 方法 行人RPE细胞原代培养并传代扩增 ,采用 Western blot法 ,分别检测角蛋白 18(cytokeratin18,CK18)、平滑肌动蛋白 (α- smooth muscle actin,α- SMA)在第 2、5、8代细胞中的表达情况。用 1∶ 4玻璃体液培养 RPE细胞6 d后 ,观察细胞形态及超微结构改变 ,采用免疫细胞化学及 Western blot法 ,检测 CK18、α- SMA表达变化。 结果 人 RPE细胞在体外培养时 ,会逐渐丧失上皮样形态 ,向成纤维样转变 ,CK18在 2代表达量中等 ,5代最高 ,而在 8代明显下降 ,而 α- SMA逐渐升高。与对照组相比 ,人玻璃体液促进 RPE细胞在形态上向成纤维样细胞转变。同时细胞内 CK18表达低于对照组 ,差异具有显著性的意义 (P<0 .0 5 ) ;α- SMA表达高于对照组 ,差异具有显著性的意义 (P<0 .0 1)。 结论 人 RPE细胞在玻璃体液与血清的作用下向具有收缩性能的间质细胞转变 ,同时细胞移行能力可能发生改变 ,结果提示玻璃体液可能促进 RPE细胞参与的 PVR
OBJECTIVE: To observe the morphological changes of human retinal pigmentepithelium (RPE) cells cultured in vitro and the changes of their cytoskeleton proteins, and to study the effect of human vitreous humor on the morphology and cytoskeletal protein expression of RPE cells. To investigate the effect of vitreous humor on proliferative vitreoretinal Regulation of retinal proliferative vitreoretinopathy (PVR). Methods Primary cultured human RPE cells were passaged and passaged. The expression of cytokeratin 18 (CK18) and α-smooth muscle actin (α-SMA) were detected by Western blot. Cell expression. The RPE cells were cultured in 1: 4 vitreous medium for 6 days. Morphological changes and ultrastructure were observed. Immunocytochemistry and Western blot were used to detect the expression of CK18 and α-SMA. Results When cultured in vitro, human RPE cells gradually lost their epithelial morphology and transformed into fibroblast-like cells. CK18 showed the highest expression in the second generation, the highest expression in the fifth generation, and a significant decrease in the eighth generation, while the α-SMA gradually increased. Compared with the control group, human vitreous humor promoted the morphological changes of RPE cells to fibroblast-like cells. Meanwhile, the expression of CK18 in cells was lower than that in the control group (P <0.05). The expression of α-SMA was higher than that in the control group (P <0.01). Conclusions Human RPE cells are transformed to contractile mesenchymal cells under the action of vitreous humor and serum, and cell migration ability may change at the same time. The results suggest that the vitreous fluid may promote the involvement of RPE cells in PVR