HIF-1α/BNIP3通路介导的糖酵解与氧诱导新生小鼠视网膜血管生成的关系  

Relationship between glycolysis mediated by HIF-1α/BNIP3 pathway and oxygen-induced retinal angiogenesis in neonatal mice

作　　者：易燕[1] 陈斐斐[1] 谭赟 杜恒[1] Yi Yan;Chen Feifei;Tan Yun;Du Heng(Dept of Pediatrics,Wuhan First Hospital(Wuhan Hospital of Integrated Traditional Chinese and Western Medicine),Wuhan 430030)

机构地区：[1]武汉市第一医院(武汉市中西医结合医院)儿科,武汉430030

出　　处：《安徽医科大学学报》2025年第2期226-233,共8页Acta Universitatis Medicinalis Anhui

基　　金：湖北省自然科学基金(编号:2019CFB401)。

摘　　要：目的基于缺氧诱导因子-1α(HIF-1α)/Bcl2/腺病毒E1B相互作用蛋白3(BNIP3)通路的糖酵解探讨氧诱导新生小鼠视网膜血管生成机制。方法将人脐静脉内皮细胞(HUVECs)分为常氧组、缺氧+si-NC组、缺氧+si-HIF-1α组和缺氧+si-HIF-1α+BNIP3组。常氧组HUVECs暴露于常氧(21%O_(2))下培养。缺氧+si-NC组、缺氧+si-HIF-1α组和缺氧+si-HIF-1α+BNIP3组用si-NC、si-HIF-1α或si-HIF-1α联合BNIP3质粒处理HUVECs 36 h,然后暴露于缺氧(1%O_(2))下培养。通过免疫荧光、代谢测量、细胞活力、划痕实验、管形成实验考察细胞线粒体自噬、糖酵解以及增殖、迁移和管形成情况。出生后第7天的C57BL/6J幼鼠随机分配到不同的治疗组:对照组、氧诱导视网膜病变(OIR)组、OIR+si-HIF-1α组和OIR+si-BNIP3组,测量新生血管形成和血管闭塞情况。结果与常氧组比较,缺氧+si-NC组HUVECs中LC3+MitoTracker+斑点数、葡萄糖摄取和乳酸释放的速率增加(P<0.001)。与缺氧+si-NC组比较,缺氧+si-HIF-1α组HUVECs中LC3+MitoTracker+斑点数、葡萄糖摄取和乳酸释放的速率降低(P<0.01)。与常氧组比较,缺氧+si-NC组HUVECs在培养第72 h的增殖活性降低(P<0.05),并且伤口愈合面积和管形成数量增加(P<0.01)。与缺氧+si-NC组比较,缺氧+si-HIF-1α组HUVECs在培养第24、48、72小时的增殖活性降低(P<0.05),伤口愈合面积、管形成数量降低(P<0.001)。BNIP3的过表达逆转了HIF-1α敲低对线粒体自噬、糖酵解以及生物学功能的影响。与OIR组比较,OIR+si-HIF-1α组和OIR+si-BNIP3组小鼠的视网膜组织中新生血管形成和血管闭塞区域减少(P<0.05)。结论HIF-1α/BNIP3信号通路在低氧条件下促进了HUVECs中线粒体自噬激活,其对于内皮功能和血管生成的调控有重要作用。Objective Based on glycolysis of hypoxia inducible factor-1α(HIF-1α)/Bcl2/adenovirus E1B interacting protein 3(BNIP3)pathway,to study the mechanism of oxygen-induced retinal angiogenesis in neonatal mice.Methods Human umbilical vein endothelial cells(HUVECs)were divided into normoxic group,hypoxia+si-NC group,hypoxia+si-HIF-1αgroup and hypoxia+si-HIF-1α+BNIP group.In normoxic group,HUVECs were exposed to normoxic(21%O_(2))and cultured.Hypoxia+si-NC group,hypoxia+si-HIF-1αgroup and hypoxia+si-HIF-1α+BNIP3 group were treated with si-NC,si-HIF-1αor si-HIF-1αcombined with BNIP3 plasmid for 36 h,and then exposed to hypoxia(1%O_(2))for culture.The autophagy,glycolysis,proliferation,migration and tube formation of mitochondria were investigated by immunofluorescence,metabolic measurement,cell viability,scratch experiment and tube formation experiment.On the 7th day after birth,C57BL/6J mice were randomly assigned to different treatment groups:control group,oxygen-induced retinopathy(OIR)group,OIR+si-HIF-1αgroup and OIR+si-BNIP group.The neovascularization and vascular occlusion were measured.Results Compared with normoxic group,the rate of LC3+MitoTracker+spots,glucose uptake and lactic acid release in HUVECs in hypoxia+si-NC group increased significantly(P<0.001).Compared with hypoxia+si-NC group,the rate of LC3+MitoTracker+spots,glucose uptake and lactic acid release in HUVECs in hypoxia+si-HIF-1αgroup decreased significantly(P<0.01).Compared with normoxic group,the proliferation activity of HUVECs in hypoxia+si-NC group decreased significantly(P<0.05),and the wound healing area and the number of tubes formed increased significantly(P<0.01).Compared with hypoxia+si-NC group,the proliferation activity of HUVECs in hypoxia+si-HIF-1αgroup decreased significantly at the 24th,48th and 72th hours of culture(P<0.05),and the wound healing area and the number of tubes formed decreased significantly(P<0.001).Overexpression of BNIP3 reversed the effects of HIF-1αknock-down on mitochondrial autophagy,glycolysis and bi

关 键 词：缺氧诱导因子-1Α 新生小鼠 视网膜血管 糖酵解 线粒体自噬 人脐静脉内皮细胞 

分 类 号：R774.1[医药卫生—眼科]