title: Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis.
abstract: Establishing a system of procedures to qualitatively and quantitatively characterize in vivo and in vitro hepatic steatosis is important for metabolic study in the liver. Here, numerous assays are described to comprehensively measure the phenotype and parameters of hepatic steatosis in mouse and hepatocyte models. Combining the physiological, histological, and biochemical methods, this system can be used to assess the progress of hepatic steatosis. In vivo, the measurements of body weight and nuclear magnetic resonance (NMR) provide a general understanding of mice in a non-invasive manner. Hematoxylin and Eosin (H&E) and Oil Red O staining determine the histological morphology and lipid deposition of liver tissue under nutrient overload conditions, such as high-fat diet feeding. Next, the total lipid contents are isolated by chloroform/methanol extraction, which are followed by a biochemical analysis for triglyceride and cholesterol. Moreover, mouse primary hepatocytes are treated with high glucose plus insulin to stimulate lipid accumulation, an efficient in vitro model to mimic diet-induced hyperglycemia and hyperinsulinemia in vivo. Then, the lipid deposition is measured by Oil Red O staining and chloroform/methanol extraction. Oil Red O staining determines intuitive hepatic steatotic phenotypes, while the lipid extraction analysis determines the parameters that can be analyzed statistically. The present protocols are of interest to scientists in the fields of fatty liver diseases, insulin resistance, and type 2 diabetes.
authors.affiliation: CAS Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Chinese Academy of Sciences.
authors.forename: Aoyuan
authors.initials: A
authors.lastname: Cui
authors.affiliation: CAS Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Chinese Academy of Sciences.
authors.forename: Zhimin
authors.initials: Z
authors.lastname: Hu
authors.affiliation: CAS Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Chinese Academy of Sciences.
authors.forename: Yamei
authors.initials: Y
authors.lastname: Han
authors.affiliation: Suzhou Niumag Analytical Instrument Corporation.
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.affiliation: CAS Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Chinese Academy of Sciences; liyu@sibs.ac.cn.
authors.forename: Yu
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D001391:Azo Compounds / Q000737:chemistry; D001835:Body Weight; D002784:Cholesterol / Q000378:metabolism; D059305:Diet, High-Fat / Q000009:adverse effects; D004195:Disease Models, Animal; D005234:Fatty Liver / Q000209:etiology*; D005947:Glucose / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism* / Q000473:pathology; D007328:Insulin / Q000378:metabolism; D008055:Lipids / Q000032:analysis*; D008099:Liver / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D008810:Mice, Inbred C57BL; D014280:Triglycerides / Q000378:metabolism
chemical_list: D001391:Azo Compounds; D007328:Insulin; D008055:Lipids; D014280:Triglycerides; D002784:Cholesterol; C011049:oil red O; D005947:Glucose