基于超高效液相色谱-质谱联用技术的慢性肾脏病骨代谢异常大鼠血清代谢组学研究  

UHPLC-MS-based serum metabolomics study in rats with chronic kidney bone disorder

作　　者：李瑾[1] 李雨静 邢晖林 邓喜文 郭明好[1] LI Jin;LI Yujing;XING Huilin;DENG Xiwen;GUO Minghao(Department of Nephrology,The First Affiliated Hospital of Xinxiang Medical University,Xinxiang,He'nan 453000,China)

机构地区：[1]新乡医学院第一附属医院肾脏内科,河南新乡453000

出　　处：《安徽医药》2024年第3期534-538,I0005,共6页Anhui Medical and Pharmaceutical Journal

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20190460);河南省重点研发与推广专项(科技攻关)项目(202102310384);河南省医学教育研究项目(Wjlx2021356)。

摘　　要：目的 通过超高效液相色谱-质谱(UHPLC-MS)联用技术探索慢性肾脏病骨代谢异常(CKD-BD)大鼠血清代谢组学变化。方法 该研究于2021年1月至2022年4月进行,选择16只8周龄SPF级SD雄性大鼠经过1周适应性喂养后,采用随机数字表法分为两组,每组8只。健康对照(NC)组:使用普通大鼠饲料喂养90 d;CKD-BD组:采用高磷高腺嘌呤饲料喂养60 d之后更换高磷饲料喂养30 d后处死,并通过生化检测及骨形态计量学方法对模型进行验证。造模成功后通过UHPLC-MS比较CKD-BD组大鼠与NC组大鼠血清代谢产物变化,通过多元统计分析等方法鉴定两组间差异代谢物,并通过生物信息学方法对差异代谢产物进行功能注释。结果 与NC组大鼠相比,CKD-BD组大鼠血肌酐、血磷显著升高,血钙显著降低(P<0.01)。骨形态计量学检测NC组大鼠骨矿化沉积率(MAR)为(1.90±0.61)μm/d,CKD-BD组大鼠MAR为(2.80±0.73)μm/d,差异有统计学意义(P<0.05)。NC组大鼠骨形成率/骨体积(BFR/BV)为(0.41±0.20)%/d,CKD-BD组大鼠BFR/BV为(1.25±0.41)%/d,差异有统计学意义(P<0.05)。代谢组学检测结果提示,与NC组相比,CKD-BD组糖基磷脂酰肌醇锚定蛋白合成等通路上调。自噬、胆汁酸合成分泌、胆固醇代谢、牛磺酸和次级牛磺酸代谢、甘油磷脂代谢等通路下调(P<0.05)。结论 通过UHPLC-MS技术比较CKD-BD大鼠与正常大鼠血清代谢产物,发现CKD疾病状态下血清代谢产物发生变化,CKD-BD组糖基磷脂酰肌醇锚定蛋白合成通路上调,与自噬相关的代谢通路下调,这些可能参与了CKD-BD发生。Objective To explore serum metabolomic changes in chronic kidney disease bone metabolism abnormalities(CKD-BD)rats by ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS).Methods The study was conducted from January 2021 to April 2022,after 1 week of adaptive feeding,sixteen 8-week-old SPF-grade SD male rats were divided into two groups by random number table method,with 8 rats in each group.Normal control(NC) group:rats were fed with normal rat chow for 90 days;CKD-BD group:rats were fed with high-phosphorus and high-adenine chow for 60 days followed by replacing the high-phosphorus chow for 30 days and then sacrificed.The model was validated by biochemical tests and bone histomorphometry.Ultra-high performance liquid chromatography-Mass Spectrum(UHPLC-MS) was used to compare the changes of serum metabolites in CKD-BD rats with those in NC rats.Multivariate statistical analysis was used to identify different metabolites between the two groups.Functional annotation of the pathways involved in the differential metabolites was analysed by bioinformatics methods.Results Compared with rats in the NC group,rats in the CKD-BD group had significantly higher blood creatinine,blood phosphorus and lower blood calcium(P < 0.01).Bone histomorphometry showed bone mineralization rate(MAR) in the NC group was(1.90±0.61) μm/day,and that of rats in the CKD-BD group was(2.80±0.73) μm/day,with a statistically significant difference(P<0.05).The bone formation rate/bone volume(BFR/BV) of the rats in the NC group was(0.41±0.20) %/day,and that of the rats in the CKD-BD group was(0.41±0.20) %/day,with a statistically significant difference(P<0.05).The results of metabolomics assay suggested that pathways such as glycosylphosphatidylinositol-anchored protein synthesis were upregulated in CKD-BD group compared with NC group.Pathways of autophagy,bile acid synthesis and secretion,cholesterol metabolism,taurine/hypotaurine metabolism,and lycerophospholipid metabolism were downregulated in CKDBD group(P < 0.05).Conc

关 键 词：慢性肾疾病-矿物质和骨代谢异常 代谢组学 超高效液相色谱-质谱 骨形态计量学 大鼠 Sprague-Dawley 

分 类 号：R692[医药卫生—泌尿科学]