眼科新进展2025，Vol.45Issue(11)：852-858,7.DOI:10.13389/j.cnki.rao.2025.0147
miR-29b-3p靶向IGF-1介导PI3K/Akt/mTOR通路对高糖诱导的人视网膜微血管内皮细胞损伤的影响
Impact of miR-29b-3p targeting IGF-1 and mediating the PI3K/Akt/mTOR pathway on high glucose-induced injury in human retinal microvascular endo-thelial cells
摘要
Abstract
Objective To investigate the effect of miR-29b-3p targeting insulin-like growth factor-1(IGF-1)on high glucose-induced injury in human retinal microvascular endothelial cells(HRMECs)and the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway.Methods HRMECs were di-vided into the following groups:control group(normal culture),high glucose(HG)group(treated with 30.0 mmol·L-1 glucose for 48 h),inhibitor-NC group,miR-29b-3p inhibitor group,miR-29b-3p inhibitor+si-NC group,miR-29b-3p inhib-itor+si-IGF-1 group(all transfected with corresponding plasmids for 24 h after HG treatment),and LY294002 group(treated with 40 μmol·L-1 LY294002 for 24 h).The expression level of miR-29b-3p was detected by quantitative real-time PCR(qRT-PCR).Cell viability was assessed using the Cell Counting Kit-8(CCK-8)assay.Intracellular reactive oxygen species(ROS)levels were measured by immunofluorescence.The expression levels of malondialdehyde(MDA),superox-ide dismutase(SOD),and glutathione peroxidase(GSH-Px)were determined using enzyme-linked immunosorbent assay(ELISA).Cell apoptosis was analyzed by flow cytometry.Autophagosome formation was observed under a transmission electron microscope.The protein expression levels related to apoptosis,autophagy,IGF-1,and the PI3K/Akt/mTOR path-way were examined by Western blot.The targeting relationship between miR-29b-3p and IGF-1 was verified by a dual-lucif-erase reporter assay.Results Compared with the Control group,the HG group showed significant increases in the ex-pression levels of miR-29b-3p,ROS,MDA,Bax,Cleaved-caspase 3,p62,the p-PI3K/PI3K ratio,the p-Akt/Akt ratio,the p-mTOR/mTOR ratio,and the cell apoptosis rate;whereas the expression levels of IGF-1,SOD,GSH-Px,the LC3Ⅱ/LC3Ⅰ ratio,cell viability,and the number of autophagosomes were significantly decreased(all P＜0.05).Compared with the in-hibitor-NC group,the miR-29b-3p inhibitor group exhibited significant decreases in the expression levels of miR-29b-3p,ROS,MDA,Bax,Cleaved-caspase 3,p62,the p-PI3K/PI3K ratio,the p-Akt/Akt ratio,the p-mTOR/mTOR ratio,and the cell apoptosis rate;while the expression levels of IGF-1,SOD,GSH-Px,the LC3Ⅱ/LC3Ⅰ ratio,cell viability,and the num-ber of autophagosomes were significantly increased(all P＜0.05).Compared with the miR-29b-3p inhibitor+si-NC group,the miR-29b-3p inhibitor+si-IGF-1 group demonstrated significant increases in the expression levels of miR-29b-3p,ROS,MDA,Bax,Cleaved-caspase 3,p62,the p-PI3K/PI3K ratio,the p-Akt/Akt ratio,the p-mTOR/mTOR ratio,and the cell apoptosis rate;whereas the expression levels of IGF-1,SOD,GSH-Px,the LC3Ⅱ/LC3Ⅰ ratio,cell viability,and the number of autophagosomes were significantly decreased(all P＜0.05).Compared with the HG group,the LY294002 group showed significant decreases in the expression levels of miR-29b-3p,ROS,MDA,Bax,Cleaved-caspase 3,p62,the p-PI3K/PI3K ratio,the p-Akt/Akt ratio,the p-mTOR/mTOR ratio,and the cell apoptosis rate;while the expression levels of IGF-1,SOD,GSH-Px,the LC3Ⅱ/LC3Ⅰ ratio,cell viability,and the number of autophagosomes were significantly increased(all P＜0.05).The dual-luciferase activity was significantly lower in cells co-transfected with IGF-1-WT and miR-29b-3p mimic compared to miR-NC cells(P＜0.05).Conclusion Inhibition of miR-29b-3p can target upregulation of IGF-1 expression levels,thereby inhibiting the PI3K/Akt/mTOR pathway and improving high glucose induced cell damage in HRMECs.关键词
miR-29b-3p/IGF-1/PI3K/Akt/mTOR通路/高糖/视网膜血管内皮细胞Key words
miR-29b-3p/IGF-1/PI3K/Akt/mTOR pathway/high glucose/human retinal microvascular endothelial cell分类
临床医学引用本文复制引用
张航烽,谢宜君,彭佳怡,王甲甲,麦涛..miR-29b-3p靶向IGF-1介导PI3K/Akt/mTOR通路对高糖诱导的人视网膜微血管内皮细胞损伤的影响[J].眼科新进展,2025,45(11):852-858,7.基金项目
国家自然科学基金-青年科学基金项目(编号:82102470) （编号:82102470）
四川省医学科研课题计划项目(编号:S23025) （编号:S23025）
