视网膜下液促使视网膜色素上皮细胞膜上蛋白激酶C变化的机制分析

Change of protein kinase C activity induced by subretinal fluid in the cell membrane of retinal pigment epithelial cells

目的:研究视网膜下液(SRF)能否引起体外培养的视网膜色素上皮(RPE)细胞浆内蛋白激酶C(PKC)的激活和转位,探讨SRF与RPE细胞中PKC信号系统变化的关系。方法:实验对象为体外培养的RPE细胞;不同的刺激因素分别在不同的时间刺激RPE细胞,通过细胞裂解和离心获取细胞浆和细胞膜蛋白粗提液,用同位素32P标记和液体闪烁计数法检测细胞浆和细胞膜PKC活性水平。结果:SRF和佛波酯(PMA)都可以激活RPE细胞浆中的PKC(c-PKC),并使其由胞浆向胞膜转位,但胞膜上PKC(m-PKC)活性峰值水平和出现的时间不同。结论:SRF可引起RPE细胞膜上PKC的活性发生变化,变化的幅度与增殖性玻璃体视网膜病变(PVR)的分级呈正相关。

Objective： To investigate whether subretinal fluid （SRF） could induce the activation and translocation of protein kinase C （PKC） in the cytoplasm of retinal pigment epithelial （RYE） cells. Methods： RPE cells cultured in vitro were stimulated with different agents at different times. The PKC activity in cytoplasm and cell membrane was measured with radioactive isotope 32P label method. Results： Both SRF and phorbol 12-myristate 13-acetate （PMA） activated PKC in the cytoplasm, and PKC in the cytoplasm was translocated into the cell membrane. The occurrence time and level of the maximal activity of PKC in the cytoplasm differed. Conclusion： SRF can induce the activation and translocation of PKC in RPE ceils, and the activity of PKC positively correlates with the degree of proliferative vitreoretinopathy.