大鼠体外循环后脑损伤与尿酸代谢的关系  

Relationship of uric acid metabolism and brain injury post-cardiopulmonary bypass in rats

作　　者：于婷婷 喻田 王海英 邓胜利 张琳 程翅 Yu Ting-Ting;Yu Tian;Wang Hai-Ying;Deng Sheng-Li;Zhang Lin;Cheng Chi(Anesthesiology Department,Affiliated Hospital of Zunyi Medical University,Zunyi,Guizhou 563000,China;Key laboratory of Anesthesia and Organ Protection of Guizhou Province,Zunyi Medical University,Zunyi,Guizhou 563000,China)

机构地区：[1]遵义医科大学附属医院麻醉科,贵州遵义563000 [2]遵义医科大学麻醉与器官保护基础研究重点实验室,贵州遵义563000

出　　处：《解放军医学杂志》2024年第10期1123-1133,共11页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(82060128);贵州省高层次人才创新创业择优资助项目[(2021)06号];遵义市科技计划[遵市科合HZ(2019)108号];遵义医科大学学术新苗培养及创新探索专项项目黔科合平台人才([2017]5733-007)。

摘　　要：目的 探究大鼠体外循环(CPB)后脑损伤与尿酸代谢的关系。方法 将健康雄性SD大鼠分为假手术组与CPB组,每组12只。假手术组仅行血管穿刺,不进行CPB转流;CPB组建立CPB模型,转流时长110 min,结束后采集脑组织。CPB前及CPB开始后均收集微透析液1 h。采用TUNEL染色检测大鼠下丘脑室旁核(PVN)区域细胞凋亡情况,实时荧光定量PCR检测大脑皮质及下丘脑中Bax mRNA的表达,Western blotting检测凋亡相关蛋白的表达。采用RNA-seq分析两组脑组织中的差异表达基因(DEGs),基因本体论(GO)分析DEGs富集的通路,使用String在线软件和Cytoscape软件构建蛋白质-蛋白质相互作用(PPI)网络并筛选出关键基因;采用液相色谱串联质谱法(LC-MS/MS)分析CPB前后PVN处微透析液的差异代谢物并进行京都基因与基因组百科全书(KEGG)富集分析。采用尿酸测定试剂盒检测下丘脑中的尿酸浓度,采用实时荧光定量PCR检测下丘脑中尿酸代谢关键酶[黄嘌呤还原酶(XDH)、腺苷脱氨酶(ADA)]及尿酸转运体[有机阴离子转运体家族蛋白1(OAT1)、有机阴离子转运体家族蛋白3(OAT3)、ATP结合盒式转运体亚家族G成员2(ABCG2)、葡萄糖转运体9(GLUT9)]基因的表达。结果 实时荧光定量PCR检测结果显示,与假手术组比较,CPB组大鼠大脑皮质及下丘脑中Bax mRNA表达量明显增高(P<0.05)。TUNEL染色结果显示,CPB组大鼠PVN区域细胞凋亡率明显增高(19.0%±5.0%vs. 7.6%±0.8%,P=0.01)。Western blotting检测结果显示,与假手术组比较,CPB组大鼠下丘脑中Bcl-2/Bax比值明显增高(P<0.05)。假手术组与CPB组大鼠脑组织中的DEGs共2829个,其中上调基因1374个,下调基因1455个。GO富集分析显示,尿酸代谢相关通路主要富集于嘌呤核苷酸代谢及生物合成、嘌呤核苷单磷酸代谢、嘌呤核苷三磷酸代谢、嘌呤核糖核苷酸代谢及生物合成、嘌呤核糖核苷单磷酸盐代谢及生物合成、嘌呤核糖核苷三磷酸Objective To investigate the relationship between uric acid metabolism and brain injury following cardiopulmonary bypass(CPB)in rats.Methods Healthy male SD rats were randomly assigned to either a Sham group or a CPB group,each comprising 12 rats.The Sham group only underwent vascular puncture and did not perform CPB conversion,while the CPB group was subjected to a CPB procedure with a perfusion duration of 110 min,and the brain tissue was collected post-procedure.Microdialysate was collected 1 h before and after CPB initiation.Apoptosis in the paraventricular nucleus(PVN)was assessed using TUNEL staining,and the expression of Bax mRNA in cerebral cortex and hypothalamus was determined via real-time quantitative PCR.Apoptosis-related protein expression was analyzed by Western blotting.Differentially expressed genes(DEGs)were identified through RNA-sequencing between brain tissues of two groups,and Gene Ontology(GO)analysis was performed to identify enriched pathways among the DEGs.Protein-protein interaction(PPI)networks were constructed using String and Cytoscape softwares to identify key genes.Liquid chromatography tandem mass spectrometry(LC-MS/MS)was employed to analyze differential metabolites in the PVN before and after CPB,with Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis constructed subsequently.Uric acid levels in the hypothalamus was measured using a uric acid assay kit,and the expression of key enzymes of uric acid metabolism[xanthine reductase(XDH),adenosine deaminase(ADA)]and uric acid transporter[organic anion transporter family protein 1(OAT1),organic anion transporter family protein 3(OAT3),ATP-binding cassette transporter subfamily G member 2(ABCG2),glucose transporter 9(GLUT9)]genes in the hypothalamus was evaluated by real-time quantitative PCR.Results Real-time quantitative PCR revealed a significant upregulation of Bax mRNA in the cerebral cortex and hypothalamus of CPB group compared to Sham group(P<0.05).TUNEL staining indicated a significantly higher apoptosis rate of

关 键 词：体外循环 脑损伤 尿酸 代谢组学 转录组学 

分 类 号：R614[医药卫生—麻醉学]