蓝光致人RPE细胞分泌VEGF及PEDF变化及与Ca2＋-PKC信号通路的关系  

The correlation between the concentrations of VEGF and PEDF and Ca2＋-PKC signaling pathways in human retinal pigment epithelial cells cultured in vitro after exposuring to blue light

作　　者：汪利敏[1] 蔡善君[1] 武志鹏[1] 宫鑫[1] 吕建平[1] 宿罡[1] 王丽丽[1] 

机构地区：[1]遵义医学院附属医院眼科贵州省眼科医院,563003

出　　处：《中华眼科杂志》2015年第11期839-843,共5页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金（81060079）;贵州省教育厅重点项目（黔教科2008034号）

摘　　要：目的 探讨蓝光照射致体外培养人RPE细胞分泌VEGF、色素上皮衍生因子（PEDF）、RPE细胞内三磷酸肌醇（IP3）和二酰甘油（DAG）的浓度变化,分析Ca2＋-蛋白激酶C（PKC）信号通路之间的相互作用.方法 实验研究.将体外培养的第4代人RPE细胞随机分为6个组,A组：无光照组;B组：蓝光光照组;C组：蓝光光照＋PKC激动剂佛波酯（PMA）组;D组：蓝光光照＋PKC抑制剂钙磷酸结合蛋白（Calphostin C）组;E组：蓝光光照＋硝苯地平组;F组：蓝光光照＋钙磷酸结合蛋白组＋硝苯地平组.用（2 000±500）lux蓝光照射体外培养人RPE细胞6h,终止培养24 h,采用ELISA测定各组RPE细胞分泌VEGF、PEDF、RPE细胞内IP3和DAG的浓度,采用流式细胞仪检测A、B及F组细胞的凋亡率.结果 A～E组的VEGF浓度分别为（584.38± 10.66）、（700.70±5.88）、（698.21±6.66）、（623.87±3.12）、（648.30±4.91） ng/L.其中B、C、E组的VEGF浓度高于A组,差异有统计学意义（P=O.002,0.002,0.016）.A～E组的PEDF浓度分别为（75.96±1.70）、（71.82±1.67）、（72.43±0.58）、（86.31＋1.35）、（93.716±1.24） μg/L.其中B、C组PEDF浓度低于A组（P=0.004,0.011）.B组和C组的VEGF/PEDF比值显著高于A组（P=0.008,0.027）,E组和D组的VEGF/PEDF比值低于B组（P=0.016,0.015）.A～E组的IP3浓度分别为（9 108.42±0.74）、（117.23±1.05）、（137.12±2.70）、（139.17±1.39）、（149.60±0.76） μg/L.B、C、D、E组IP3浓度均高于A组（P=0.003,0.007,0.000,0.000）,并且C、D、E组IP3浓度均高于B组（P=0.011,0.000,0.000）.A～E组的DAG浓度分别为（995.47±13.61）、（1070.10±10.07）、（1046.39±7.90）、（1041.12±9.76）、（1273.13±10.89） pmol/L.B、C、D、E组DAG浓度均高于A组（P=0.000,0.000,0.000,0.000）,与B组比较,组DAG的浓度增高（P=0.000）,而C、D组DAG浓度降低（P=0.021,0.007）.A、B和F组的细胞凋亡率分别是（10.26±1.87）％、（25.06±2.66）％和�Objective To investigate the concentrations of vascular endothelial growth factor （VEGF）, pigment epithelium-derived factor （PEDF）, inositol triphosphate （IP3） and diacylglycerol （DAG） in human retinal pigment epithelium （RPE） cells after exposuring to blue light, and to explore the relationship with Ca2 ＋-PKC signaling pathways, to evaluate the role of Ca2＋-PKC signaling pathways of blue-light irradiation induced apoptosis in RPE cells.Methods The fourth generation human RPE cells in vitro were exposured to blue light （2000± 5001ux） for 6 hours, 24 hours prolongation of post-exposure culture.The concentrations of VEGF, PEDF, IP3 and DAG were assayed by enzyme linked immunosorbent assay （ELISA）.Cells were randomly divided into 6 groups, group A（eontrol）, group B（exposure to blue light）, group C（exposure to blue light ＋ PMA）, group D（exposure to blue light＋Calphostin C）, group E（exposure to blue light ＋ Nifedipine）, group F （exposure to blue light＋ Calphostin C＋ Nifedipine）.Flow cytometry was used to detect the apoptosis rate of human RPE cells in A, B and F group.Results Comparing with group A （584.38± 10.66）, the concentration of VEGF in group B （700.70±5.88）, groupC （698.21±6.66）and group E （648.30±4.91） was higher, the difference was statistically significant （P=0.002, 0.002, 0.016）.Comparing with group B （700.70±5.88）, the concentration of VEGF in Group D （623.87±3.12） and E （648.30±4.91） was lower （P=0.001, 0.002）.Comparing with group A （75.96 ± 1.70）, the concentration of PEDF in Group B （71.82± 1.67）and C （72.43±0.58）was lower （P=0.004,0.011）, but the concentration of PEDF in Group D （86.31 ± 1.35） and E （93.72± 1.24） was higher （P=0.000,0.000）.Comparing with group B （71.82± 1.67）, the concentration of PEDF in Group D （86.31 ± 1.35） and E （93.72± 1.24） was higher （P=0.000, 0.000）.Comparing with group A （7.70±0.29）, the ratio of VEGF to PEDF i

关 键 词：光刺激 视网膜色素上皮 血管内皮生长因子A 眼蛋白质类 神经生长因 子类 细胞凋亡 

分 类 号：R774.1[医药卫生—眼科]