基于UHPLC-QE-MS的高原鼠兔肾脏差异代谢物及低氧适应机制分析  

Analysis of Kidney Differential Metabolites and Hypoxia Adaptation Mechanism of Plateau Pikas Based on UHPLC-QE-MS

作　　者：何雨昕 白振忠[1,2] 薛华 郭子旭 曹学锋[1,2] HE Yuxin;BAI Zhenzhong;XUE Hua;GUO Zixu;CAO Xuefeng(Medical College of Qinghai University,Xining 810001,China;High Altitude Medical Research Center,Qinghai University,Xining 810001,China)

机构地区：[1]青海大学医学院,西宁810001 [2]青海大学高原医学研究中心,西宁810001

出　　处：《实验动物与比较医学》2025年第1期3-12,共10页Laboratory Animal and Comparative Medicine

基　　金：青海省昆仑英才·高端创新人才·培养领军项目(K9924072)。

摘　　要：目的运用超高效液相色谱-四极杆静电场轨道阱质谱(ultra-high-performance liquid chromatography coupled with quadrupole electrostatic field orbital trap-mass spectrometry,UHPLC-QE-MS)非靶向代谢组学分析法探讨不同海拔高原鼠兔肾脏低氧适应性代谢变化的潜在机制。方法捕捉青海省果洛藏族自治州玛多县星宿海地区海拔4360 m(MD组)和青海省海北藏族自治州门源地区海拔2900 m(MY组)处的高原鼠兔各10只,经麻醉后采集血清样本,经安死术后采集肾脏样本,分别进行一般生理生化指标测定和代谢组学分析。其中一部分血清样本用于血液学分析,另一部分用于血气分析,剩余部分检测生化指标。肾组织样本中代谢物提取后,进行UHPLC-QE-MS分析,运用代谢组学主成分分析(principal component analysis,PCA)和正交偏最小二乘判别分析(orthogonal partial least squares discriminant analysis,OPLS-DA)方法分析差异代谢物,筛选标准为变量投影重要度(variable Importance in the projection,VIP)>1.5且倍数变化(fold change,FC)>1.5或VIP>1.5且FC<1/1.5。利用相关性分析热力图、差异显著性分析火山图、信号通路识别气泡图和矩形图分别分析差异代谢物及相关信号通路。结果MD组高原鼠兔的红细胞计数、葡萄糖、尿素氮、尿酸和同型半胱氨酸含量高于MY组,而血红蛋白、血细胞比容、肌酐和二氧化碳结合力低于MY组,这表明不同海拔的高原鼠兔血液携氧能力存在明显差异。经主成分模式识别分析和OPLS-DA置换检验显示,MD组和MY组高原鼠兔肾脏代谢物具有明显的聚类型分布(R^(2)Y=0.930,Q^(2)=0.655)。按照筛选标准,并经数据库比对发现,不同海拔高原鼠兔的肾脏代谢物差异分子有46个,其中MD组高原鼠兔的蟾蜍二烯羟酸内酯、腺苷、腺嘌呤、薯蓣皂苷、盐酸小檗碱、鼠尾草酚和虾青素表达水平均显著升高(VIP>1.5,P<0.05),花生四烯酸、组胺和香豆素水平显著下降Objective To explore the potential mechanisms of hypoxic adaptive metabolic changes in the kidneys of plateau pikas at different altitudes using non-targeted metabolomics analysis via ultra-high-performance liquid chromatography coupled with quadrupole electrostatic field orbital trap-mass spectrometry(UHPLC-QE-MS).Methods 10 plateau pikas were captured at an altitude of 4360 m in Xingxiuhai area,Maduo County,Guoluo Tibetan Autonomous Prefecture,Qinghai Province(MD group),and 10 plateau pikas were captured at an altitude of 2900 m in Menyuan area,Haibei Tibetan Autonomous Prefecture,Qinghai Province(MY group).After anesthesia,serum samples were collected,and kidney samples were collected after euthanasia.General physiological and biochemical indicators were measured and metabolomics analysis was performed.Part of the serum samples was used for hematology analysis,another part for blood gas analysis,and the remaining part for biochemical indicator detection.Metabolites were extracted from the kidney tissue samples and then analyzed using UHPLC-QE-MS.Differential metabolites were analyzed using metabolomics principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA),with screening criteria set as variable importance in projection(VIP)>1.5 and fold change(FC)>1.5,or VIP>1.5 and FC<1/1.5.Correlation analysis heatmaps,significance analysis volcano plots,signaling pathway recognition bubble charts,and rectangular graphs were used for the analysis of differential metabolites and related signaling pathways.Results The red blood cell count,glucose,urea nitrogen,uric acid,and homocysteine levels in the MD group plateau pikas were higher than those in the MY group,while hemoglobin,hematocrit,creatinine,and carbon dioxide combining power were lower than those in the MY group.This indicated a significant difference in the blood oxygen-carrying capacity of plateau pikas at different altitudes.The principal component pattern recognition analyses,and OPLS-DA permutation test showed that t

关 键 词：高原低氧 高原鼠兔 肾脏 超高效液相色谱-四极杆静电场轨道阱质谱 代谢通路 

分 类 号：Q95-33[生物学—动物学] R-332[医药卫生]