Proteomic Profiling of Liver and Plasma in Chronic Ethanol Feeding Model of Hepatic Alcohol Dehydrogenase‐Deficient Deer Mice

• Published in: Alcoholism, clinical and experimental research clinical and experimental research, 2017-10, Vol.41 (10), p.1675-1685
• Main Authors: Bhopale, Kamlesh K., Amer, Samir M., Kaphalia, Lata, Soman, Kizhake V., Wiktorowicz, John E., Shakeel Ansari, Ghulam A., Kaphalia, Bhupendra S.
• Format: Article
• Language: English
• Subjects: Abuse; Actin; Alcohol; Alcohol abuse; Alcohol dehydrogenase; Alcohol Dehydrogenase - deficiency; Alcohol Dehydrogenase - genetics; Alcohol use; Aldolase; Animals; Apolipoprotein E; ATP synthase; Carbamoyl phosphate; Carbohydrate metabolism; Carbohydrates; Carbonic anhydrase; Carbonic anhydrases; Cirrhosis; Clusterin; Creatine; Creatine kinase; Cytochrome b; Cytokeratin; Deer Mice; Dehydrogenases; Drug abuse; Electrophoresis; Endoplasmic reticulum; Ethanol; Ethanol - administration & dosage; Ethanol - toxicity; Fibrosis; Gel electrophoresis; Hepatitis; Histology; Isoforms; Laminin; Liver; Liver - drug effects; Liver - metabolism; Liver - pathology; Liver cirrhosis; Liver diseases; Liver Diseases, Alcoholic - genetics; Liver Diseases, Alcoholic - metabolism; Liver Diseases, Alcoholic - pathology; Lymphocytes; Lymphocytes T; Male; Mice; Oxidation; Peromyscus; Plasma; Plasma proteins; Proteins; Proteomics; Proteomics - methods; Rodents
• ISSN: 0145-6008; 1530-0277
• DOI: 10.1111/acer.13470

Background Chronic alcohol abuse, a major risk factor for such diseases as hepatitis and cirrhosis, impairs hepatic alcohol dehydrogenase (ADH; key ethanol [EtOH]‐metabolizing enzyme). Therefore, differentially altered hepatic and plasma proteomes were identified in chronic EtOH feeding model of hepatic ADH‐deficient (ADH−) deer mice to understand the metabolic basis of alcoholic liver disease (ALD). Methods ADH− deer mice were fed 3.5 g% EtOH via Lieber–DeCarli liquid diet daily for 3 months and histology of the liver assessed. Liver and plasma proteins were separated by 2‐dimensional gel electrophoresis. The proteins differentially expressed were identified by matrix‐assisted laser desorption ionization‐time of flight mass spectrometry. Results Histology of the liver showed panlobular steatosis and infiltration of T lymphocytes. Using the criteria of ≥1.5 for fold change (p‐value ≤0.05) with expectation value (E ≤10−3) and protein score (≥64), 18 proteins in the livers and 5 in the plasma of EtOH‐fed mice were differentially expressed and identified. Prolyl 4‐hydroxylase, cytochrome b‐5, endo A cytokeratin, ATP synthase, heat‐shock 70 kD proteins, enoyl CoA hydratase, stress‐70 protein, peroxiredoxin 1, and ornithine carbamoyl transferase were up‐regulated in the livers. However, carbonic anhydrase 3, mitochondrial ATP synthase, aldolase 2, actin γ, laminin receptor, and carbamoyl phosphate synthase were down‐regulated. Contrary to the increased expression of creatine kinase M‐type, a decreased expression of serine protease inhibitor A3A precursor, sulfated glycoprotein‐2 (clusterin), and apolipoprotein E isoforms were found in the plasma of EtOH group. Conclusions Chronic EtOH feeding in ADH− deer mice causes steatosis and infiltration of T lymphocytes in the livers along with increased expression of proteins involved in endoplasmic reticulum (ER) stress, fibrosis, fatty acid β oxidation and biogenesis, and decreased expression of proteins involved in ATP synthesis, carbohydrate metabolism, in cell regulation and architecture. Reduced expression of various carrier proteins as found in the plasma of EtOH group has a biomarker potential.