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. 2015 Oct;149(4):1030-41.e6.
doi: 10.1053/j.gastro.2015.06.009. Epub 2015 Jun 20.

Fat-Specific Protein 27/CIDEC Promotes Development of Alcoholic Steatohepatitis in Mice and Humans

Ming-Jiang Xu  1 , Yan Cai  2 , Hua Wang  3 , José Altamirano  4 , Binxia Chang  5 , Adeline Bertola  5 , Gemma Odena  6 , Jim Lu  7 , Naoki Tanaka  8 , Kimihiko Matsusue  9 , Tsutomu Matsubara  8 , Partha Mukhopadhyay  10 , Shioko Kimura  8 , Pal Pacher  10 , Frank J Gonzalez  8 , Ramon Bataller  11 , Bin Gao  12
Affiliations

Fat-Specific Protein 27/CIDEC Promotes Development of Alcoholic Steatohepatitis in Mice and Humans

Ming-Jiang Xu et al. Gastroenterology. 2015 Oct.

Abstract

Background & aims: Alcoholic steatohepatitis (ASH) is the progressive form of alcoholic liver disease and may lead to cirrhosis and hepatocellular carcinoma. We studied mouse models and human tissues to identify molecules associated with ASH progression and focused on the mouse fat-specific protein 27 (FSP-27)/human cell death-inducing DFF45-like effector C (CIDEC) protein, which is expressed in white adipose tissues and promotes formation of fat droplets.

Methods: C57BL/6N mice or mice with hepatocyte-specific disruption of Fsp27 (Fsp27(Hep-/-) mice) were fed the Lieber-Decarli ethanol liquid diet (5% ethanol) for 10 days to 12 weeks, followed by 1 or multiple binges of ethanol (5 or 6 g/kg) during the chronic feeding. Some mice were given an inhibitor (GW9662) of peroxisome proliferator-activated receptor γ (PPARG). Adenoviral vectors were used to express transgenes or small hairpin (sh) RNAs in cultured hepatocytes and in mice. Liver tissue samples were collected from ethanol-fed mice or from 31 patients with alcoholic hepatitis (AH) with biopsy-proved ASH and analyzed histologically and immunohistochemically and by transcriptome, immunoblotting, and real-time PCR analyses.

Results: Chronic-plus-binge ethanol feeding of mice, which mimics the drinking pattern of patients with AH, produced severe ASH and mild fibrosis. Microarray analyses revealed similar alterations in expression of many hepatic genes in ethanol-fed mice and humans with ASH, including up-regulation of mouse Fsp27 (also called Cidec) and human CIDEC. Fsp27(Hep-/-) mice and mice given injections of adenovirus-Fsp27shRNA had markedly reduced ASH following chronic-plus-binge ethanol feeding. Inhibition of PPARG and cyclic AMP-responsive element binding protein H (CREBH) prevented the increases in Fsp27α and FSP27β mRNAs, respectively, and reduced liver injury in this chronic-plus-binge ethanol feeding model. Overexpression of FSP27 and ethanol exposure had synergistic effects in inducing production of mitochondrial reactive oxygen species and damage to hepatocytes in mice. Hepatic CIDEC mRNA expression was increased in patients with AH and correlated with the degree of hepatic steatosis and disease severity including mortality.

Conclusions: In mice, chronic-plus-binge ethanol feeding induces ASH that mimics some histological and molecular features observed in patients with AH. Hepatic expression of FSP27/CIDEC is highly up-regulated in mice following chronic-plus-binge ethanol feeding and in patients with AH; this up-regulation contributes to alcohol-induced liver damage.

Keywords: Endoplasmic Reticulum; Inflammation; Lipid Metabolism; Translational Research.

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Conflict of interest statement

Conflicts of interest: All authors disclose no conflicts.

Figures

Figure 1
Figure 1. Chronic-plus-binge ethanol feeding induces ASH
A–C, Eight- to ten-week-old male C57BL6N mice were fed a liquid diet containing 5% ethanol for 10 days or for 4 to 12 weeks-plus-one binge (E10d+1B, E4-12w+1B) or with multiple binges (E8w+nB), for 8 weeks without a binge (E8w), or pair-fed (P8w); the mice were euthanized at 9 hours post-binge. (A) Representative images of livers. (B) Representative images of H&E staining. (C) Serum ALT and AST levels. Values represent the mean ± SEM (n=5–30 mice per group). *P<0.05 in comparison with E8w; #P<0.05 as indicated.
Figure 2
Figure 2. Characterization of chronic-plus-binge feeding model (E8w+1B and E8w+nB)
(A) Representative photographs (200x) of various stainings of liver tissues. (B) Neutrophil infiltration per field (100×), number of inflammatory foci per 100 fields, Sirius red positive area, and hepatic expression of collagens were determined. (C) Real-time quantitative PCR analyses of pro-inflammatory genes. *P<0.05, **P<0.01, ***P<0.001 in E8w+1B and E8w+nB in comparison with P8w or E8w. #P<0.05 as indicated. (n=5–8 mice per group).
Figure 3
Figure 3. Chronic-plus-binge feeding upregulates hepatic Fsp27α and Fsp27β expression through PPARγ- and CREBH-N-dependent mechanisms, respectively
Mice from different feeding models were used. (A, B, C) Real-time PCR analyses of liver tissues. (D) Western blot analyses of liver tissues. The density of the protein bands was quantified on the right panel. (E, F) Mice were fed E8w+1B, and vehicle or the PPAR-γ inhibitor GW9662 was administered i.p. 15 minutes before the binge treatment, or infected with Ad-control-shRNA or Ad-shCREBH 5 days before ethanol gavage. Hepatic Fsp27α and Fsp27β mRNA levels were determined by real-time PCR analyses. *P<0.05, **P<0.01, ***P<0.001; #P<0.05 as indicated. (n=3–12 mice per group).
Figure 4
Figure 4. Inhibition of hepatic Fsp27 ameliorates chronic-plus-binge ethanol-induced liver injury
(A–C) Mice were fed E8w+1B and administered Ad-control-shRNA or Ad-shFsp27 for the final 5 days. Real-time PCR analyses, serum ALT and AST levels, and representative H&E staining of the liver are shown. (D–F) Fsp27Hep−/− and WT mice were fed E8w+1B or E8w+nB. Mice were euthanized 9 hours post the last binge. Serum ALT and AST levels, liver triglyceride levels, representative H&E staining (E8w+1B), and real-time PCR analyses are shown. *P<0.05, **P<0.01, ***P<0.001 as indicated. (n=6–8 mice per group).
Figure 5
Figure 5. Ethanol and FSP27 synergistically induce mitochondrial ROS production and hepatocyte death
(A, B, C) Mice were fed E8w+1B and administered Ad-control-shRNA or Ad-shFsp27 for the final 5 days, and were euthanized 9 hours post binge. Liver tissues were subjected to MDA staining (panel A) and western blotting for 4-HNE (panel B). Cytoplasm and mitochondria were isolated from fresh liver tissues and subjected to western blot analysis (panel C). (D, E) Hepatocytes were infected with Adcontrol or Ad-Fsp27-HA for 36 hours, then incubated with ethanol for 6 hours. Cytoplasm and mitochondria were isolated for western blotting (panel D). Mitochondrial ROS generation was measured (panel E). ROS production was also measured in hepatocytes from WT and Fsp27Hep−/− mice (panel E). (F) Ad-control-vector- or Ad-Fsp27-HA-treated hepatocytes were incubated with various concentrations of ethanol for 6 hours, ALT levels in the supernatant were measured. *P<0.05, **P<0.01.
Figure 6
Figure 6. Hepatic CIDEC mRNA levels are increased and positively correlated with the severity of disease in AH patients
(A) Real-time PCR analyses of liver samples from patients and from healthy controls. (B) Correlation between CIDEC mRNA and the degree of steatosis. (C) Correlation between CIDEC mRNA and MELD score, ABIC score, and HVPG. (D) Hepatic CIDEC mRNA levels in alive and deceased AH patients. (E) Hepatic CIDEC mRNA level and 90-day mortality. P values are presented in the figure.
Figure 7
Figure 7. A proposed model for a critical role of FSP27 in chronic-plus-binge ethanol-induced ASH
ERS: endoplasmic reticulum stress; LD: lipid droplet.
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References

    1. Gao B, Bataller R. Alcoholic liver disease: pathogenesis and new therapeutic targets. Gastroenterology. 2011;141:1572–1585. - PMC - PubMed
    1. Tsukamoto H, Lu SC. Current concepts in the pathogenesis of alcoholic liver injury. FASEB J. 2001;15:1335–1349. - PubMed
    1. Williams JA, Manley S, Ding WX. New advances in molecular mechanisms and emerging therapeutic targets in alcoholic liver diseases. World J Gastroenterol. 2014;20:12908–12933. - PMC - PubMed
    1. Lucey MR, Mathurin P, Morgan TR. Alcoholic hepatitis. N Engl J Med. 2009;360:2758–2769. - PubMed
    1. Singal AK, Kamath PS, Gores GJ, et al. Alcoholic hepatitis: current challenges and future directions. Clin Gastroenterol Hepatol. 2014;12:555–564. quiz e31–2. - PMC - PubMed

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