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

发布时间：2018-11-27 10:24

【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(P0.01);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(P0.05)。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。
[Abstract]:Objective to observe the effects of p44 / 42 MAPK kinase inhibitor PD98059 and PI3K/Akt kinase inhibitor LY294002 on the expression of vascular endothelial growth factor (vascular endothelial growth factor, in human retinal pigment epithelial cell line ARPE-19 induced by non-enzymatic glycosylation end product (advanced glycation end-products,AGE). VEGF). Methods ARPE-19 cells were treated with different concentrations of AGE and AGE for different time. The expression of extracellular VEGF protein in ARPE-19 cells was detected by ELISA method. The expression of phosphorylated p44 / 42 and Akt was detected by immunocytofluorescence assay. ARPE-19 cells were pretreated with PD98059 and / or LY294002, then treated with AGE. The extracellular VEGF level, phosphorylated p44 / 42 and Akt activity were measured. Results with the increase of AGE concentration or action time, the expression of VEGF in ARPE-19 cells increased, and the AGE concentration reached the peak at 100mg L-1 or 8 h, and the expression level was (304.55 卤20.51) ng L -1, respectively. (293.54 卤13.98) ng L -1, while p44 / 42 and Akt signaling molecules were activated. After pretreatment with PD98059 or LY294002, the expression of p44 / 42 and Akt was low, and the expression of extracellular VEGF induced by AGE was also decreased in a dose-dependent manner with PD98059 and LY294002. The expression of VEGF was the lowest when 20 渭 mol L-1 PD98059 and 30 渭 mol L-1 LY294002 were used. The expression of VEGF was (57.35 卤5.29) pg L -1 and (81.51 卤8.10) pg L -1, respectively. However, the expression of VEGF was positively correlated with the concentration of AGE and the time of action (P0.01). The expression of VEGF induced by AGE was not related to the time of AGE treatment with PD98059 and LY294002 (P0.05). Conclusion p44/42MAPK and PI3K/Akt signaling pathway is an important intracellular signal pathway for AGE to induce VEGF expression in ARPE-19 cells. PD98059 and LY294002 can inhibit AGE induced VEGF expression in human RPE cells.
【作者单位】：云南省第一人民医院眼科;中南大学湘雅二医院眼科;
【基金】：云南省应用基础研究计划项目(编号:2011FZ285)~~
【分类号】：R587.2;R774.1

【参考文献】