p42/p44MAPK通路在高糖诱导的hRPE细胞VEGF表达中的作用  

Role of p42 /p44MAPK signal transduction pathway in expression of VEGF induced by elevated glucose concentration in cultured hRPE cells

作　　者：张春侠[1] 胡建民[2] 庄铭忠[2] 

机构地区：[1]济南市明水眼科医院,中国山东省章丘市1250200 [2]福建医科大学附属第二医院眼科,中国福建省泉州市2362000

出　　处：《国际眼科杂志》2014年第8期1382-1385,共4页International Eye Science

摘　　要：目的:探讨p42/p44丝裂原活化蛋白激酶(mitogenactivated protein kinases,MAPK)信号转导通路在高糖诱导的人视网膜色素上皮(human retinal pigment epithelium,hRPE)细胞血管内皮生长因子(vascular endothelial growth factor,VEGF)表达中的作用。方法:采用hRPE细胞株,将细胞分为正常对照组(5.6mmol/L葡萄糖)、高糖对照组(15,20,30mmol/L葡萄糖)、PD98059处理组(20μmol/L p42/p44MAPK高效选择性抑制剂PD98059处理hRPE细胞)和溶剂二甲基亚砜对照组(dimethyl sulfoxide,DMSO组)。应用逆转录PCR(RTPCR)技术检测VEGF及色素上皮细胞衍生因子(pigment epithelium derived factor,PEDF)mRNA的表达。应用酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)技术检测细胞上清液中VEGF蛋白的表达结果:高糖作用下VEGF mRNA和蛋白表达显著增高,PD98059处理组VEGF mRNA和蛋白表达受到抑制,且VEGF mRNA/PEDF mRNA比值较高糖组显著降低。结论:p42/p44MAPK信号转导通路可能参与了高糖引起的hRPE细胞VEGF的表达。AIM： To study p42/p44 mitogen- activated protein kinases（ MAPK） signal transduction pathway effect on vascular endothelial growth factor（ VEGF） expression induced by elevated glucose concentration in cultured human retinal pigment epithelium（hRPE）.METHODS：hRPE cells were cultured and divided into four groups： normal glucose group（ NG）（5. 6mmol /L）,high glucose group（ HG1：15mmol /L D- glucose,HG2：20mmol /L D- glucose,HG3： 30 mmol /L D- glucose）,PD98059 group： hRPE cells were treated by an efficient and selective inhibitor PD98059（ 20μ mol /L） of p42/p44 MAPK signal transduction pathway and solvent dimethyl sulfoxide group（DMSO group）. The expression of VEGF and pigment epithelium derived factor（PEDF） mRNA was detected by RT- PCR. VEGF protein expression in cultured hRPE supernatants was detected by enzyme-linked immumosorbent assay（ELISA）.RUSULTS：VEGF mRNA and protein expression induced by elevated glucose concentration increased significantly.VEGF mRNA and protein expression were restrained in PD98059 group. Ratio of（ VEGF/β- actine） /（ PEDF/β-actine）in PD98059 group decreased significantly compare with that in high glucose group. CONCLUSION： p42/p44 MAPK signal transduction pathway might play a part in VEGF expression induced by elevated glucose concentration in cultured hRPE cells.

关 键 词：视网膜色素上皮细胞 高浓度葡萄糖 血管内皮生长因子 色素上皮源性衍生因子 丝裂原活化蛋白激酶 

分 类 号：R774.1[医药卫生—眼科]