培养人视网膜色素上皮细胞在PVR患者视网膜下液作用下c-fos和c-jun的表达

目的 观察培养的人视网膜色素上皮细胞(human retinal pigment epithelium，hRPE)在不同程度增生性玻璃体视网膜病变(proliferative vitreoretinopathy，PVR)患者视网膜下液(subretinal fluid，SRF)作用下细胞增生与c—fos和c—jun的表达。方法 不同程度PVR患者SRF作用于培养的hRPE细胞后利用细胞计数和MTT比色实验观察SRF对培养hRPE细胞的作用，同时采用SP法在不同时间点作兔抗人c—fos多抗和兔抗人c—jun多抗的免疫组织化学染色。结果 PVR患者SRF作用组与对照组相比可明显促进培养hRPE细胞的增殖，同时实验组hRPE细胞c—fos和c—jun均表达增高，视网膜下液作用lh后，Fos蛋白抗体染色阳性，细胞核呈明显棕黄色染色；3h表达达高峰，持续约12h；视网膜下液作用lh后，Jun蛋白抗体染色阳性，细胞核呈明显棕黄色染色；6h表达达高峰，持续约48h，对照组hRPE细胞(未经SRF作用仅加入等量PBS)Fos蛋白抗体和Jun蛋白抗体染色均为阴性。结论 PVR患者SRF中含有能诱导c—fos和c—jun等“立早基因”转录的细胞外生长或分化因子；c—fos和c—jun的表达是SRF促进RPE细胞的增殖过程中一个非常重要的早期分子事件。

Effects of subretinal fluid on expression of c-fos and c-jun in cultured human retinal pigment epithelium

Objective To observe the expression of c fos and c jun in cultured human retinal pigment epithelium (hRPE) after treatment with subretinal fluid (SRF) from different grades of proliferative vitreoretinopathy (PVR).Methods The effect of SRF from different grades of PVR on cultured hRPE was detected by the methods of cell counting and MTT assay. SP techniques were applied for immunohistochemical staining of cultured hRPE with rabbit anti human c fos antibodies and rabbit anti human c jun antibodies.Results All SRF samples showed a stimulating effect on cultured hRPE in a certain degree compared with the control group.While all SRF samples could stimulate the expression of c fos and c jun in cultured hRPE.The positive staining was mottled in nucleus.There was significant difference between the SRF treated group and the control group.Conclusion SRF from PVR includes some growth factor that can induce the expression of c fos and c jun in cultured hRPE.We can conclude that hRPE cell proliferation is under fos and jun control.