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

摘要:

目的：探讨p42/p44丝裂原活化蛋白激酶(mitogen-activated 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(RT-PCR)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。

Abstract:

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 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/p44MAPK signal transduction pathway might play a part in VEGF expression induced by elevated glucose concentration in cultured hRPE cells.