【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。更多还原

【Abstract】 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 receptor δ（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 PPARδ and 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 PPARδ receptor 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）; the expression level of ANGPTL4 in the C20DC treatment group was higher than that in the control group, and the difference was statistically significant（P<0.05）. Conclusion C20DC can promote the expression of ANGPTL4 protein by binding to PPARδ and thus increase the proliferation and migration of retinal related cells（HRECs and ARPE-19 cells）. Its mechanism may be related to the increased angiogenesis in retinopathy of prematurity.更多还原