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, HG3:30mmol/L D - glucose ), PD98059 group: hRPE cells were treated by an efficient and selective inhibitor PD98059 (20μmol/L) of p42/p44MAPK 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