PDLIM1通过下调自噬抑制大鼠视网膜新生血管形成的机制研究

PDLIM1 suppresses retinal neovascularization in rats by downregulating autophagy

目的探究PDLIM1对高糖培养的大鼠视网膜毛细血管内皮细胞(retinal capillary endothelial cell,RCEC)自噬通路和血管形成的影响及机制。设计实验研究。研究对象大鼠视网膜毛细血管内皮细胞。方法将RCEC随机分为四组,正常对照组(NG),高糖组(HG),转染对照组(HG+NC组),PDLIM1过表达组(HG+ad-rPDLIM1组)。采用血管形成实验、划痕实验检测血管生成情况和迁移能力,蛋白质免疫印迹和实时荧光定量PCR检测细胞中PDLIM1及微管相关蛋白轻链3(LC3)、苄氯素1(Beclin-1)的蛋白和mRNA表达水平。主要指标细胞成管数,细胞迁移量以及LC3、Beclin-1抗体的蛋白、mRNA水平。结果HG组细胞成管数、迁移量高于对照组,PDLIM1在HG组中的表达降低,而LC3、Beclin-1的表达升高;过表达PDLIM1后,与HG组相比,细胞成管数(2.33±1.15)个、细胞迁移量(12.33±1.08)%显著降低(t=9.59,P<0.001;t=22.39,P<0.001),LC3、Beclin-1表达明显减少(t=22.39,P<0.001;t=11.1,P<0.001)。结论PDLIM1可能通过下调自噬作用抑制大鼠的视网膜新生血管的生成。(眼科,2024,33:290-294)

Objetive To investigate the effect of PDLIM1 on autophagy and vascular formation of rats retinal capillary endothelial cells(RCECs)with high glucose culture.Design Experiment study.Participants Rats retinal capillary endothelial cells.Methods The RCECs were randomly divided into normal glucose(NG),high glucose(HG),high glucose+negative control(HG+NC)and HG+ad-rPDLIM1 groups.Tubule formation test,scratch test were used to determine the ability of cell tube formation and proliferation.The protein and mRNA expression levels of PDLIM1,LC3 and Beclin-1 were verified by Western blotting and RT-qPCR.Main Outcome Measures The number of cell tubes,the amount of cell migration and the protein and mRNA levels of LC3 and Beclin-1 antibodies.Results The number of tube formation and migration of cells in the HG group were higher than those in NG group,and the expression of PDLIM1 decreased in the HG group,while the expression of LC3 and Beclin-1 increased.After overexpression of PDLIM1,compared with the HG group,the number of tube formation(2.33±1.15)and cell migration(12.33±1.08)%significantly decreased(t=9.59,P<0.001;t=22.39,P<0.001),and the expression of LC3 and Beclin-1 significantly decreased(t=22.39,P<0.001;t=11.1,P<0.001).Conclusions PDLIM1 may inhibit retinal neovascularization in rats through downregulating autophagy.(Ophthalmol CHN,2024,33:290-294)