PD98059和LY294002对非酶糖基化终产物诱导人RPE细胞表达VEGF的影响  

Effects of PD98059 and LY294002 on VEGF expression in human retinal pigment epithelium induced by advanced glycation end products

作　　者：杨晓春[1] 许建彪[1] 唐罗生[2] 

机构地区：[1]云南省第一人民医院眼科,云南省昆明市650032 [2]中南大学湘雅二医院眼科,湖南省长沙市410000

出　　处：《眼科新进展》2014年第9期817-820,825,共5页Recent Advances in Ophthalmology

基　　金：云南省应用基础研究计划项目(编号:2011FZ285)~~

摘　　要：目的观察p44/42 MAPK激酶抑制剂PD98059和PI3K/Akt激酶抑制剂LY294002对非酶糖基化终产物(advanced glycation end-products,AGE)诱导的人视网膜色素上皮细胞株ARPE-19细胞表达血管内皮生长因子(vascular endothelial growth factor,VEGF)的影响。方法分别用不同浓度AGE和AGE作用不同时间干预ARPE-19细胞,ELISA方法检测ARPE-19细胞外VEGF蛋白表达水平,免疫细胞荧光法检测磷酸化p44/42和Akt的表达;以PD98059和(或)LY294002预处理ARPE-19细胞后再用AGE干预,并测定ARPE-19细胞外VEGF水平、磷酸化p44/42和Akt活性。结果随着AGE浓度或作用时间增加,ARPE-19细胞表达VEGF外增加,AGE浓度为100mg·L-1或作用8h时达高峰,表达水平分别为(304.55±20.51)ng·L-1、(293.54±13.98)ng·L-1,同时p44/42和Akt信号分子激活。使用PD98059或LY294002预处理后,p44/42和Akt低表达,AGE诱导的ARPE-19细胞外VEGF表达也降低,并与PD98059和LY294002呈剂量依赖性,当使用20μmol·L-1 PD98059和30μmol·L-1 LY294002时,VEGF的表达最低,分别为(57.35±5.29)pg·L-1、(81.51±8.10)pg·L-1,但VEGF的表达量仍与AGE浓度和作用时间呈正相关(P<0.01);同时使用PD98059和LY294002预处理,AGE诱导的VEGF表达与AGE的作用时间无关(P>0.05)。结论 p44/42MAPK和PI3K/Akt信号通路是AGE诱导ARPE-19细胞表达VEGF的重要细胞内信号通路,PD98059和LY294002可抑制AGE诱导的人RPE细胞外VEGF表达。Objective To investigate the effects of PD98059 （p44/42 MAPK inhibitor） and LY294002 （PI3K/Akt inhibitor） on expression of vascular endothelial growth factor （VEGF） in ARPE-19 cells induced by advanced glycation end products （AGE）. Methods ARPE-19 cells were treated with different doses of AGE for certain time or with certain dose for different time, then the amount of VEGF was measured by ELISA, and expression of MAPK and Akt was tested by immunofluorescence. Cells were pre-treated with PD98059 and/or LY294002, then with AGE. The amount of VEGF, MAPK and Akt were measured too. Results The amount of VEGF expression in ARPE-19 cells increased with the increasing of AGE concentration or incubation time, peaked at 100 mg · L^ -1 or 8 hours, which were （ 304.55 ±20.51 ） ng· L^-1 , （ 293.54 ± 13.98） ng· L ^-1, respectively. Meanwhile, pM4/42 MAPK and PI3K/Akt signal pathways were activated. The amount of phosphorylated p44/42 or Akt decreased on dose dependent when the cells were treated with PDgS059 or LY294002 firstly. The minimum amount of VEGF were（57.35 ±5.29）pg · L^ -1 and （81.51 ±8.10）pg · L^-1 after being intervened by 20 μmol · L^-1 PD98059 and 30 μmol · L^ -1 LY294002, respectively. The amount of VEGF decreased remarkably but still remained positive correlation with concentration and incubation time of AGE（P 〈 0.01 ）. However,there was no correlation when the cells were treated with both PD98059 and LY294002 firstly （ all P 〉 0. 05 ）. Conclusion Both 1〉44/42 MAPK and PI3K/Akt signal pathways play important roles in the expression of VEGF in ARPE-19 cells induced by AGE ,which can be depressed by PD98059 and LY294002.

关 键 词：糖基化终产物 丝裂素活化蛋白激酶 磷脂酰肌醇-3激酶 色素上皮 信号通路 

分 类 号：R77[医药卫生—眼科]