二十烷二酸通过结合PPARδ介导ANGPTL4表达增多对人视网膜血管内皮细胞增殖和迁移的影响  

Effects of eicosanoic acid on proliferation and migration of human retinal vascular endothelial cells by mediating increased expression of angiopoietin-like protein 4 after binding to peroxisome proliferator-activated receptor δ

作　　者：杨宇航 祁慧 董利军 范梓欣 陆小凤 王明良 余震 雷和田 张国明[1] YANG Yuhang;QI Hui;DONG Lijun;FAN Zixin;LU Xiaofeng;WANG Mingliang;YU Zhen;LEI Hetian;ZHANG Guoming(Shenzhen Eye Hospital,Shenzhen 518040,Guangdong Province,China;School of Chemistry and Environmental Engineering,Shenzhen University,Shenzhen 518055,Guangdong Province,China)

机构地区：[1]深圳市眼科医院,广东省深圳市518040 [2]深圳大学化学与环境工程学院,广东省深圳市518055

出　　处：《眼科新进展》2024年第9期679-685,共7页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助项目(编号:82271103);广东省基础与应用基础研究基金项目(编号:2022A1515012326);广东省高水平临床重点专科(深圳市配套建设经费)资助(编号:SZGSP014);深圳市科技计划资助(编号:JCYJ20220530153415035);深圳市“医疗卫生三名工程”项目资助(编号:SZSM202311018);深圳市医学研究专项资金(编号:C2301005);深圳市医学重点学科建设经费资助(编号:SZXK038)。

摘　　要：目的 研究二十烷二酸(C20DC)对人视网膜内皮细胞(HREC)增殖、迁移能力的影响,并探讨其作用机制。方法 寻找C20DC干预人视网膜色素上皮-19(ARPE-19)细胞与HREC的最佳工作浓度,分别为30 mg·L^(-1)与25 mg·L^(-1)。将HREC分为C20DC处理组(C20DC干预HREC)和对照组[二甲基亚砜(DMSO)干预HREC],通过细胞增殖和迁移实验观察C20DC对HREC迁移及增殖能力的影响;采用分子对接方法模拟C20DC与PPARδ的结合能力;将ARPE-19细胞设为C20DC+ARPE-19组(C20DC干预ARPE-19细胞)与DMSO+ARPE-19组(DMSO干预ARPE-19细胞),采用Western blot检测ARPE-19细胞中PPARδ和血管生成素样蛋白4(ANGPTL4)蛋白,以及HREC中ANGPTL4蛋白表达水平;采用ELISA定量分析ARPE-19细胞与HREC中ANGPTL4蛋白表达水平。结果 与对照组相比,C20DC处理组细胞增殖、迁移能力均显著提高(均为P<0.05),且可与PPARδ结合稳定(结合能为-7.2 kcal·mol^(-1));Western blot检测结果提示,C20DC+ARPE-19组细胞中ANGPTL4蛋白表达量高于DMSO+ARPE-19组,差异有统计学意义(P<0.05),两组PPARδ受体蛋白表达量比较,差异无统计学意义(P>0.05);C20DC处理组细胞中ANGPTL4蛋白表达量高于对照组,差异有统计学意义(P<0.05)。ELISA定量分析检测结果显示,C20DC+ARPE-19组细胞中ANGPTL4表达量高于DMSO+ARPE-19组(P<0.001);C20DC处理组细胞中ANGPTL4的表达量高于对照组,差异有统计学意义(P<0.05)。结论 C20DC可结合PPARδ促进ANGPTL4蛋白的表达,导致视网膜相关细胞(HREC与ARPE-19细胞)增殖与迁移能力增强,其作用机制可能与早产儿视网膜病变新生血管生成增多有关。Objective To investigate the effects of eicosanoic acid(C20DC)on the proliferation and migration of human retinal endothelial cells(HRECs)and its mechanism.Methods The optimal working concentration of C20DC in human retinal pigment epithelium 19(ARPE-19)cells and HRECs was determined as 30 mg·L^(-1) and 25 mg·L^(-1),respectively.HRECs were divided into the C20DC treatment group(HRECs treated with C20DC)and the control group[HRECs treated with dimethyl sulfoxide(DMSO)].The effects of C20DC on the migration and proliferation of HRECs were detected by cell proliferation and migration experiments.The molecular docking method was used to simulate the binding ability of C20DC to peroxisome proliferator-activated receptorδ(PPARδ).ARPE-19 cells were divided into the C20DC+ARPE-19 group(ARPE-19 cells treated with C20DC)and the DMSO+ARPE-19 group(ARPE-19 cells treated with DMSO).The expression levels of PPARδand angiopoietin-like protein 4(ANGPTL4)in ARPE-19 cells and ANGPTL4 protein in HRECs were detected using Western blot.The ANGPTL4 protein expression levels in ARPE-19 cells and HRECs were quantitatively analyzed using enzyme-linked immunosorbent assay(ELISA).Results Compared with the control group,the proliferation and migration of cells in the C20DC treatment group significantly increased(both P<0.05),and C20DC could stably bind to PPARδ(binding energy:-7.20 kcal·mol^(-1)).Western blot showed that the expression level of ANGPTL4 protein in the C20DC+ARPE-19 group was higher than that in the DMSO+ARPE-19 group,and the difference was statistically significant(P<0.05);there was no statistically significant difference in the expression level of PPARδreceptor protein between the two groups(P>0.05).The expression level of ANGPTL4 protein in the C20DC treatment group was higher than that in the control group,and the difference was statistically significant(P<0.05).ELISA quantitative analysis showed that the expression level of ANGPTL4 in the C20DC+ARPE-19 group was higher than that in the DMSO+ARPE-19 group(P<0.001)

关 键 词：二十烷二酸(C20DC) 早产儿视网膜病变 血管生成素样蛋白4 过氧化物酶体增殖物激活受体 脂肪酸 视网膜微血管内皮细胞 

分 类 号：R774[医药卫生—眼科]