氧诱导视网膜病变模型小鼠肾组织代谢组学研究  

Metabolomics study of kidney tissue in a mouse model of oxygen-induced retinopathy

作　　者：董利军 祁慧 杨宇航 毛星星 张国明[1] 张少冲 雷和田 Dong Lijun;Qi Hui;Yang Yuhang;Mao Xingxing;Zhang Guoming;Zhang Shaochong;Lei Hetian(Shenzhen Eye Hospital,Jinan University,Shenzhen Eye Institute,Shenzhen 518040,China)

机构地区：[1]深圳市眼科医院、暨南大学附属深圳眼科医院、深圳市眼病防治研究所,深圳518040

出　　处：《中华实验眼科杂志》2024年第1期19-28,共10页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金(82070989);深圳市"医疗卫生三名工程"项目(SZSM202011015)。

摘　　要：目的探讨高氧环境对氧诱导视网膜病变(OIR)模型小鼠肾脏代谢物的影响,了解病理性视网膜血管新生和肾损伤之间的潜在机制。方法采用随机数字表法将16只健康SPF级C57/B6J新生小鼠随机分为OIR组与正常对照组,每组8只。小鼠自出生后标准饲养至第7天(P7),OIR组小鼠和母鼠置于(75±2)%的高氧箱中饲养至P12,然后正常饲养;正常对照组一直在正常环境下饲养。各组小鼠在饲养P17时采用二氧化碳安乐死,取视网膜组织铺片并行血管异凝集素(IB4)染色,观察视网膜血管形态、中央无灌注区及病理性新生血管情况;另取小鼠肾组织进行液相色谱-质谱分析,取其对应小鼠的全血进行抗凝处理,通过离心沉淀,获得不含细胞成分的血浆,对血浆进行靶向代谢组学分析。使用代谢组学数据处理软件Progenesis QI v2.3对质谱信息进行解析,用无监督的主成分分析及正交偏最小二乘法分析(OPLS-DA)来区分各组间代谢轮廓的总体差异,比较2个组间代谢物的倍数变化。以变量权重值>1且P<0.05为条件筛选出差异代谢物。基于KEGG数据库对差异代谢物进行代谢通路富集分析。结果视网膜铺片IB4染色结果显示,P17时正常对照组小鼠视网膜血管分布均匀;OIR组小鼠视网膜周边血管迂曲、紊乱,中央可见大面积无灌注区域形成,在视网膜无灌注区和血管区交界处形成大量新生血管簇,呈强荧光染色。OIR组小鼠视网膜无灌注区相对面积为(25.16±3.50)%,明显大于正常对照组的(0.63±0.30)%,差异有统计学意义(t=12.07,P<0.001)。OPLS-DA模型参数R2X cum、解释率R2Y cum、和预测率Q2 cum分别为0.578、0.978和0.857,表明OPLS-DA模型具有较好的预测能力。共筛选鉴定到26个主要的差异代谢物,其中上调表达17个,下调表达9个,包括甘油磷脂类化合物[PC 20∶4(5Z,8Z,11Z,14Z)/0∶0、PC 22∶6(4Z,7Z,10Z,13Z,16Z,19Z)/0∶0、PC 14∶1(9Z)/20∶2(11Z,14Z)、PE P-18∶0/20�Objective To explore the effects of hyperoxic environments on renal metabolites to understand the potential mechanisms that contribute to pathologic retinal vascular neovascularization and renal injury through metabolomic studies in a mouse model of oxygen-induced retinopathy(OIR)model.Methods Sixteen C57/B6J mice pups born to day 7(P7)were randomly and equally divided into an OIR model group and a normal control group using a randomized numerical table of mother mice.Mice were reared standardly from birth until day 7(P7),then mice and their mother mice in the OIR group were placed in a hyperoxic(75±2)%chamber until day 12(P12)and then reared normally.Mice in the normal control group were reared normally throughout.Mice in two groups were killed by carbon dioxide euthanasia on postnatal day 17(P17).The mice retinal wholemount from the two groups were made and stained with isolectin B4(IB4)to observe the morphology of retinal vessels,central non-perfusion area and pathological neovascularization.The kidney tissue of P17 mice was analyzed by liquid chromatograph mass spectrometer.After anticoagulant treatment,the whole blood of mice was centrifuged and precipitated,and the obtained plasma without cellular components was analyzed by targeted metabonomics.Mass spectral information was interpreted using metabolomics data processing software Progenesis QI v2.3.Overall differences in metabolic profiles were distinguished by unsupervised principal component analysis and orthogonal partial least squares analysis(OPLS-DA).The fold change and P values of metabolites were compared between the two groups.The variable importance of projection value>1 and P value<0.05 was used to screen out differential metabolites.Metabolic pathway enrichment analysis of differential metabolites was performed based on the KEGG database.The feeding and use of animals were strictly in accordance with the requirements of the Ethics Committee of Jinan University,and the research protocol was reviewed and approved by the Ethics Committee of Jinan

关 键 词：氧诱导视网膜病变 小鼠 肾脏 液相色谱-质谱 代谢产物分析 早产儿视网膜病变 

分 类 号：R774.1[医药卫生—眼科] R-332[医药卫生—临床医学]