基于HPLC-MS/MS技术的酮病奶牛血清脂质代谢物分析  

Analysis of serum lipid metabolites in ketotic dairy cows based on HPLC-MS/MS technology

作　　者：李卓 常仁旭 孙旭东[1] 徐闯[1] LI Zhuo;CHANG Renxu;SUN Xudong;XU Chuang(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang163319,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《中国兽医学报》2024年第4期802-807,共6页Chinese Journal of Veterinary Science

基　　金：黑龙江省应用技术研究与开发计划资助项目(GA20B201)。

摘　　要：对25头荷斯坦奶牛进行初次筛选,包含13头疑似酮病和12头疑似健康奶牛。依据临床症状和血液酮体浓度将其分为临床型酮病组(血液中BHBA≥3.0mmol/L,NEFA>0.5mmol/L,且表现消瘦、食欲减退)及健康组(血液中BHBA<1.2mmol/L,NEFA<0.4mmol/L,临床检查无异常表现)。选取胎次相近、产奶量和体况相似的临床型酮病奶牛、健康奶牛各5头。采集奶牛全血到促凝管中,使用高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)检测酮病奶牛血清中的异常脂质分子。选取P<0.05,VIP>1的脂质分子。结果显示:共筛选出18个与酮病相关的差异代谢物,与健康组相比,酮病组其中有5种磷脂酰胆碱(phosphatidylcholine,PC)、4种胆固醇酯(cholesterol esters,CE)、2种鞘磷脂(sphingomyelin,SM)、2种溶血磷脂酰胆碱(lyso-phosphatidyl choline,LPC)、2种甘油三酯(triglyceride,TAG)降低;有3种游离脂肪酸(free fat acid,FFA)升高。结果表明,这些异常的脂质可能是酮病奶牛的新型标志物。To investigate novel differential metabolites of serum lipids in dairy cows with ketosis,so as to provide new diagnostic techniques and theoretical support for the pathogenesis of ketosis.The 25early postpartum Holstein cows including 13suspected clinical ketotic cows,and 12healthy cows were selected.The cows were classified as clinical ketotic cows(blood BHB concentrations≥3mmol/L,NEFA>0.5 mmol/L,rapid loss of live weight,reduce in feed intake)and healthy(blood BHB concentrations<1.2mmol/L,NEFA<0.4mmol/L,no obvious clinical symptoms)according to clinical symptoms and blood BHB concentrations.Five clinical ketotic cows and 5 healthy cows with similar calving date milk production and body weight were enrolled in this study.Blood samples were collected into procoagulant tubes via coccygeal venipuncture before feeding,and abnormal lipid molecules of serum were detected by high-performance liquid chromatographytandem mass spectrometry(HPLC-MS/MS).The lipid molecule with P<0.05,VIP>1were selected.The results shown that a total of 18differential metabolites associated with clinical ketotic cows.Compared with healthy group,five kinds of phosphatidylcholine(PC),4kinds of cholesterol esters(CE),2kinds of sphingomyelin,SM,2kinds of lyso-phosphatidyl choline(LPC),and 2kinds of lyso-phosphatidyl choline(LPC)was increased in the ketosis group.Two kinds of triglycerides(TAG)were decreased,while three free fat acids(FFA)were increased in dairy cows with ketosis compared with healthy cows.These above abnormal lipids may serve as new biomarkers for ketosis in dairy cows.

关 键 词：奶牛 围产期 酮病 脂质组学 HPLC-MS/MS 

分 类 号：S852.3[农业科学—基础兽医学] S856[农业科学—兽医学]