基于液质联用技术对早期高脂血症金黄地鼠肝脏鞘脂的靶向代谢组学研究

Targeted Metabonomics of Sphingolipids in Liver Tissue from Early Stage Hyperlipidemia Hamsters Using Liquid Chromatography-tandem Mass Spectrometry

目的对高脂血症金黄地鼠肝匀浆中鞘脂进行相对定量的靶向代谢组学研究。方法18只SPF级叙利亚金黄地鼠随机分为正常对照组和高脂血症模型组,饲喂6周后进行血液生化检测,取肝脏进行HE染色,观察两组动物血液生化指标和组织病理学变化。基于液质联用技术,采用主成分分析(PCA)和偏最小二乘法分析(PLS-DA)进行模式识别,对鞘脂种类进行相对定量分析。结果高脂血症模型组TG、TC、LDL-C、HDL-C水平明显高于正常对照组,均具有统计学差异。肝组织结构紊乱,肝细胞大片气球样变性,灶性小泡性脂肪变,肝细胞肥大致肝窦不明显,枯否细胞增生,肝小叶内可见炎细胞聚集灶,汇管区轻度炎细胞浸润及纤维组织增生。21种鞘脂,包括鞘磷脂(sphingomyelin,SM)、神经酰胺(ceramide,Cer)和葡糖基神经酰胺(glucosyl-ceramide,Glc-cer)在高脂血症仓鼠肝脏出现异常改变。其中,9种Cer上调,4种Glc-cer下调,8种SM中有3种上调,其余5种下调。结论早期高脂血症金黄地鼠的围绕神经酰胺的鞘脂的代谢途径被破坏,可能触模型组的胰岛素抵抗和炎症反应,导致肝脏损伤,影响血脂代谢。基于LC-MS的鞘脂途径的靶向代谢组学研究为了解高脂血症的生物学机理提供了更多有用的信息。

In this study,a relative quantitative analysis of the sphingolipids in liver homogenate from hyperlipidemia hamsters was carried out by the liquid chromatography-tandem mass spectrometry(LC-MS).Principal component analysis(PCA)and Partial least-squares to latent structure-discriminant analysis(PLS-DA)were used for data analysis and obvious difference was observed from normal and hyperlipidemia group.Twenty one sphingolipids including sphingomyelins(SM),ceramides(Cer),and glucosyl-ceramide(Glc-cer)showed abnormal change in the liver of hyperlipidemic hamsters.Among them 9 ceramides were up-regulated,4 Glc-cers were down-regulated,while three of eight SMs were up regulated,other five of them were down-regulated.The result indicate the compounds of the SMs-Cers-glycosphingolipids(GLP)pathway were disrupted in the model group and it may trigger insulin resistance and inflammation which cause damage on liver in hyperlipidemia.The targeted metabonomic study of sphingolipid pathway based on LC-MS provided more useful information to understand the pathology of hyperlipidemia disease.