中文摘要：

目的：研究视网膜下液（SRF）能否引起体外培养的人视网膜色素上皮（RPE）和神经胶质（RG）细胞发生增殖及在其增殖过程中是否出现蛋白激酶C的激活和转位，来探讨RPE和RG细胞增殖与其细胞中PKC信号系统变化的关系以及PKC抑制剂的作用。 方法：实验对象为体外培养的RPE和RG细胞；刺激因素为提取PVR分级是B、C级两级患者的SRF和来自角膜移植后的供体眼球所提供的正常玻璃体成分；PKC特异激活剂佛波酯（PMA）为阳性对照；DMEM培养液作为空白对照。用3H-胸腺嘧啶脱氧核苷（3H-TdR）掺入法测定各自的RPE和RG细胞增殖情况。用B、C级SRF、正常玻璃体成分、PMA、DMEM培养液分别在不同的时间刺激RPE和RG细胞，通过细胞裂解和离心获取细胞质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质和细胞膜PKC活性水平。选用PKC抑制剂地喹氯铵预处理各组细胞后，再分别观察各组RPE和RG细胞中PKC活性表达水平及增殖情况。 结果：用B、C级SRF和PMA处理过的RPE细胞出现高增殖；SRF和PMA都可以激活RPE和RG细胞质中的PKC，并使其由胞质向胞膜转位，但SRF作用于RPE和RG细胞时，胞膜上PKC活性峰值出现的时间较PMA明显延长。其中，B级SRF作用于RPE和RG细胞时，胞膜上PKC活性峰值出现的时间较C级长且峰值低，增殖程度也低；正常玻璃体成分和DMEM培养液组没有出现PKC活性变化和高增殖。用PKC特异抑制剂预处理各组细胞后，没有出现PKC活性和细胞增殖的改变，组间无显著性差异，P＞0.05。 结论：SRF可促进RPE和RG细胞增殖，RPE和RG细胞中的PKC是以激活和转位方式参与细胞增殖的过程；使用PKC特异抑制剂可阻止此过程发生。

英文摘要：

AIM： To study the effect of the subretinal fluid （SRF） on proliferation of retinal pigment epithelium （RPE） cells and retinal glial （RG） cells and associated activation and translocation of protein kinase C （PKC） as well as the application of PKC inhibitor. MTEHODS： RPE and RG cells were disintegrated to obtain PKC activity of cytoplasm and cellular membrane after being treated by the subretinal fluid （SRF） from the different stages of PVR patients （grade B and C） or being treated with PKC specific activator [phorbol-12-myris-tate-13-acetate （PMA）] or normal vitreous or DMEM culture medium. PKC activity in cytoplasm and cellular membrane was measured using radioactive isotope 32P labeling in a specific reaction of phosphorylation on PKC substrate. In addition, the PKC inhibitor, dequalinium chloride, was used to pretreat the RPE and RG cells before the cells exposed to SRF or PMA or normal vitreous. 3H-TdR （tritiated thymidine） was used to measure the levels of proliferation of RPE and RG cells with or without the activation and translocation. RESULTS： SRF and PMA promoted the proliferation of RPE and RG cells. SRF and PMA activated PKC in the cytoplasm of RPE and RG cells and the activated cytoplasm PKC translocated to the cellular membrane of RPE or RG cells. The cell proliferation or PKC activation or translocation were not equally active in RPE as in RG cells. However, PKC inhibitor which attenuated the cell proliferation did not show significant difference on inhibition of RPE and RG cell proliferation. （P 〉0.05）. CONCLUSION： SRF can lead to the activation and translocation of PKC in RPE and RG cells, which promote the proliferation of RPE and RG cells. Dequalinium chloride can inhibit PKC activation and translocation hence slow down the cells proliferation.