Wee Hai Ning, Lee Lye Siang, Han Sharon Hong Yu, Zhou Jin, Yen Paul Michael, Ching Jianhong
Cardiovascular and Metabolic Disorders Programme, Duke-NUS Medical School, Singapore, Singapore.
KK Research Centre, KK Women's and Children's Hospital, Singapore, Singapore.
Bio Protoc. 2023 Jul 5;13(13):e4773. doi: 10.21769/BioProtoc.4773.
Non-alcoholic steatohepatitis (NASH) is a condition characterized by inflammation and hepatic injury/fibrosis caused by the accumulation of ectopic fats in the liver. Recent advances in lipidomics have allowed the identification and characterization of lipid species and have revealed signature patterns of various diseases. Here, we describe a lipidomics workflow to assess the lipid profiles of liver homogenates taken from a NASH mouse model. The protocol described below was used to extract and analyze the metabolites from the livers of mice with NASH by liquid chromatography-mass spectrometry (LC-MS); however, it can be applied to other tissue homogenate samples. Using this method, over 1,000 species of lipids from five classes can be analyzed in a single run on the LC-MS. Also, partial elucidation of the identity of neutral lipid (triacylglycerides and diacylglycerides) aliphatic chains can be performed with this simple LC-MS setup. Key features Over 1,000 lipid species (sphingolipids, cholesteryl esters, neutral lipids, phospholipids, fatty acids) are analyzed in one run. Analysis of liver lipids in non-alcoholic steatohepatitis (NASH) mouse model. Normal-phase chromatography coupled to a triple quadrupole mass spectrometer.
非酒精性脂肪性肝炎（NASH）是一种由肝脏中异位脂肪堆积引起炎症和肝损伤/纤维化的病症。脂质组学的最新进展使得脂质种类得以鉴定和表征，并揭示了各种疾病的特征模式。在此，我们描述了一种脂质组学工作流程，用于评估取自NASH小鼠模型的肝脏匀浆的脂质谱。以下所述方案用于通过液相色谱 - 质谱联用（LC-MS）从患有NASH的小鼠肝脏中提取和分析代谢物；然而，它也可应用于其他组织匀浆样品。使用此方法，在LC-MS上单次运行可分析来自五类的1000多种脂质。此外，利用这种简单的LC-MS设置可以对中性脂质（三酰甘油和二酰甘油）脂肪链的身份进行部分阐明。关键特性单次运行分析超过1000种脂质种类（鞘脂、胆固醇酯、中性脂质、磷脂、脂肪酸）。在非酒精性脂肪性肝炎（NASH）小鼠模型中分析肝脏脂质。正相色谱与三重四极杆质谱仪联用。
