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Knockout of the Gsta4 Gene in Male Mice Leads to an Altered Pattern of Hepatic Protein Carbonylation and Enhanced Inflammation Following Chronic Consumption of an Ethanol Diet
Author(s) -
Shearn Colin T.,
Pulliam Casey F.,
Pedersen Kim,
Meredith Kyle,
Mercer Kelly E.,
Saba Laura M.,
Orlicky David J.,
Ronis Martin J.,
Petersen Dennis R.
Publication year - 2018
Publication title -
alcoholism: clinical and experimental research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.267
H-Index - 153
eISSN - 1530-0277
pISSN - 0145-6008
DOI - 10.1111/acer.13766
Subject(s) - steatosis , endocrinology , medicine , protein carbonylation , liver injury , alcoholic liver disease , chemistry , tumor necrosis factor alpha , biology , biochemistry , microbiology and biotechnology , lipid peroxidation , oxidative stress , cirrhosis
Background Glutathione S‐transferase A4‐4 ( GSTA 4) is a key enzyme for removal of toxic lipid peroxidation products such as 4‐hydroxynonenal (4‐ HNE ). In this study, we examined the potential role of GSTA 4 on protein carbonylation and progression of alcoholic liver disease by examining the development of liver injury in male wild‐type (WT) SV/J mice and SV /J mice lacking functional GSTA 4 ( GSTA 4 −/− mice). Methods Adult male WT and GSTA 4 −/− mice were fed chow ( N = 10 to 12) or high‐fat Lieber‐DeCarli liquid diets containing up to 28% calories as ethanol (EtOH) ( N = 18 to 20) for 116 days. At the end of the study, half of the EtOH‐fed mice were acutely challenged with an EtOH binge (3 g/kg given intragastrically) 12 hours before sacrifice. Carbonylation of liver proteins was assessed by immunohistochemical staining for 4‐ HNE adduction and by comprehensive liquid chromatography–tandem mass spectrometry ( LC ‐ MS / MS ) of purified carbonylated proteins. Results Chronic Et OH intake significantly increased hepatic 4‐ HNE adduction and protein carbonylation, including carbonylation of ribosomal proteins. Et OH intake also resulted in steatosis and increased serum alanine aminotransferase. Hepatic infiltration with B cells, T cells, and neutrophils and mRNA expression of pro‐inflammatory cytokines tumor necrosis factor (TNF) α and interferon (IFN) γ was modest in WT mice. However, an EtOH binge increased hepatic necrosis, hepatic cell proliferation, and expression of TNF α mRNA ( p < 0.05). Et OH treatment of GSTA 4 −/− mice increased B‐cell infiltration and increased mRNA expression of TNF α and IFN γ and of matrix remodeling markers MMP 9, MMP 13, and Col 1A1 ( p < 0.05). GSTA 4 −/− mice exhibited panlobular rather than periportal distribution of 4‐ HNE ‐adducted proteins and increased overall 4‐ HNE staining after Et OH binge. Comprehensive LC‐MS of carbonylated proteins identified 1,022 proteins of which 189 were unique to the GSTA 4 −/− group. Conclusions These data suggest long‐term adaptation to Et OH in WT mice does not occur in GSTA 4 −/− mice. Products of lipid peroxidation appear to play a role in inflammatory responses due to Et OH . And Et OH effects on B‐cell infiltration and autoimmune responses may be secondary to formation of carbonyl adducts.