Role of CHOP in Hepatic Apoptosis in the Murine Model of Intragastric Ethanol Feeding

Overview

Journal Alcohol Clin Exp Res

Specialty Psychiatry

Date 2005 Sep 1

PMID 16131858

Citations 86

Authors

Cheng Ji

Ruty Mehrian-Shai

Christine Chan

Ya-Hsuan Hsu

Neil Kaplowitz

Affiliations

Soon will be listed here.

Abstract

Background: CHOP is a transcriptional regulator involved in apoptosis caused by endoplasmic reticulum (ER) stress. We previously reported that CHOP as well as other ER stress response genes is induced in the liver of a murine model of intragastric ethanol feeding. This study was undertaken to determine the role of CHOP in hepatocellular apoptosis and liver injury in this model.

Methods: CHOP wild-type (+/+) mice and CHOP null (-/-) mice were fed alcohol for four weeks with glucose as control. Hematoxylin-eosin staining, TUNEL, and caspase 3 staining of liver tissues were performed for assessment of fatty liver, necroinflammation, and apoptosis. Total RNA was extracted for microarray and reverse transcription-PCR analyses, and proteins were used for western blotting.

Results: Significant increased liver/body ratio, steatosis, liver triglyceride levels, and plasma homocysteine concentrations were observed in alcohol-fed mice as compared with controls in both genotypes. There was no significant difference between wild-type and CHOP null (-/-) mice in the parameters related to fatty liver. Alcohol-induced increased serum alanine aminotransferase levels and necroinflammatory foci were not significantly reduced in CHOP null (-/-) mice. However, apoptosis was present in alcohol-fed wild-type mice but virtually absent in alcohol-fed CHOP null (-/-) mice. The ER stress response indicated by increased Grp78 mRNA was observed in both types of mice fed alcohol. Of 12,423 transcripts analyzed for >or= two-fold changes, several related to apoptosis were influenced by CHOP: Gadd45 and cathepsin B were up-regulated in ethanol-fed wild-type mice but not in CHOP null (-/-) mice, whereas Jun D and Bcl-xL were down-regulated in ethanol-fed wild-type mice but not in ethanol-fed CHOP null (-/-) mice.

Conclusions: CHOP null (-/-) mice have remarkable absence of hepatocellular apoptosis in response to alcohol feeding but no protection against hyperhomocysteinemia, ER stress, and fatty liver. Thus, CHOP up-regulation occurs downstream of and contributes to one manifestation of ER stress, namely, apoptosis. Microarray studies confirmed by PCR analysis and western blotting indicate that genes affected by CHOP are both proapoptotic and antiapoptotic and CHOP induction by ethanol may tip the balance of cell survival and death toward apoptosis.

Citing Articles

Association of serum and local GRP78 and CHOP expressions with disease progression in patients with non-traumatic osteonecrosis of femoral head.

Zhang P, Ye Q, Zhu W, Zhao Y, Zhu H, Wei B J Orthop Surg Res. 2025; 20(1):108.

PMID: 39881366 PMC: 11776197. DOI: 10.1186/s13018-025-05541-5.

Oral supplementation of choline attenuates the development of alcohol-related liver disease (ALD).

Sanchez V, Baumann A, Kromm F, Yergaliyev T, Brandt A, Scholda J Mol Med. 2024; 30(1):181.

PMID: 39425011 PMC: 11488139. DOI: 10.1186/s10020-024-00950-4.

Characterizing alcohol-related and metabolic dysfunction-associated steatotic liver disease cirrhosis via fibrotic pattern analysis.

Fukushima M, Miyaaki H, Nakao Y, Sasaki R, Haraguchi M, Takahashi K Sci Rep. 2024; 14(1):23679.

PMID: 39390024 PMC: 11466976. DOI: 10.1038/s41598-024-73739-4.

How to Use the Cuprizone Model to Study De- and Remyelination.

Kipp M Int J Mol Sci. 2024; 25(3).

PMID: 38338724 PMC: 10855335. DOI: 10.3390/ijms25031445.

Lumpy Skin Disease Virus Infection Activates Autophagy and Endoplasmic Reticulum Stress-Related Cell Apoptosis in Primary Bovine Embryonic Fibroblast Cells.

Tan J, Liu Y, Li W, Zhang Y, Chen G, Fang Y Microorganisms. 2023; 11(8).

PMID: 37630443 PMC: 10456949. DOI: 10.3390/microorganisms11081883.

References

Baroni G, Marucci L, Benedetti A, Mancini R, JEZEQUEL A, Orlandi F . Chronic ethanol feeding increases apoptosis and cell proliferation in rat liver. J Hepatol. 1994; 20(4):508-13. DOI: 10.1016/s0168-8278(05)80498-2. View

Boise L, Gonzalez-Garcia M, Postema C, Ding L, LINDSTEN T, Turka L . bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell. 1993; 74(4):597-608. DOI: 10.1016/0092-8674(93)90508-n. View

Kearsey J, Coates P, Prescott A, Warbrick E, Hall P . Gadd45 is a nuclear cell cycle regulated protein which interacts with p21Cip1. Oncogene. 1995; 11(9):1675-83. View

BARAK A, Beckenhauer H, Tuma D . Betaine, ethanol, and the liver: a review. Alcohol. 1996; 13(4):395-8. DOI: 10.1016/0741-8329(96)00030-4. View

Kaplowitz N, Tsukamoto H . Oxidative stress and liver disease. Prog Liver Dis. 1996; 14:131-59. View

Tsujimoto Y, Shimizu S, Eguchi Y, Kamiike W, Matsuda H . Bcl-2 and Bcl-xL block apoptosis as well as necrosis: possible involvement of common mediators in apoptotic and necrotic signal transduction pathways. Leukemia. 1997; 11 Suppl 3:380-2. View

Zinszner H, Kuroda M, Wang X, Batchvarova N, Lightfoot R, Remotti H . CHOP is implicated in programmed cell death in response to impaired function of the endoplasmic reticulum. Genes Dev. 1998; 12(7):982-95. PMC: 316680. DOI: 10.1101/gad.12.7.982. View

Sok J, Wang X, Batchvarova N, Kuroda M, Harding H, Ron D . CHOP-Dependent stress-inducible expression of a novel form of carbonic anhydrase VI. Mol Cell Biol. 1998; 19(1):495-504. PMC: 83907. DOI: 10.1128/MCB.19.1.495. View

Fawcett T, Martindale J, Guyton K, Hai T, Holbrook N . Complexes containing activating transcription factor (ATF)/cAMP-responsive-element-binding protein (CREB) interact with the CCAAT/enhancer-binding protein (C/EBP)-ATF composite site to regulate Gadd153 expression during the stress response. Biochem J. 1999; 339 ( Pt 1):135-41. PMC: 1220137. View

10.

Artuso M, Esteve A, Bresil H, VUILLAUME M, Hall J . The role of the Ataxia telangiectasia gene in the p53, WAF1/CIP1(p21)- and GADD45-mediated response to DNA damage produced by ionising radiation. Oncogene. 1995; 11(8):1427-35. View

11.

Ron D, Habener J . CHOP, a novel developmentally regulated nuclear protein that dimerizes with transcription factors C/EBP and LAP and functions as a dominant-negative inhibitor of gene transcription. Genes Dev. 1992; 6(3):439-53. DOI: 10.1101/gad.6.3.439. View

12.

Benedetti A, Brunelli E, Risicato R, Cilluffo T, JEZEQUEL A, Orlandi F . Subcellular changes and apoptosis induced by ethanol in rat liver. J Hepatol. 1988; 6(2):137-43. DOI: 10.1016/s0168-8278(88)80024-2. View

13.

Guicciardi M, Deussing J, Miyoshi H, Bronk S, Svingen P, Peters C . Cathepsin B contributes to TNF-alpha-mediated hepatocyte apoptosis by promoting mitochondrial release of cytochrome c. J Clin Invest. 2000; 106(9):1127-37. PMC: 301415. DOI: 10.1172/JCI9914. View

14.

Avila M, Berasain C, Torres L, Martin-Duce A, Corrales F, Yang H . Reduced mRNA abundance of the main enzymes involved in methionine metabolism in human liver cirrhosis and hepatocellular carcinoma. J Hepatol. 2000; 33(6):907-14. DOI: 10.1016/s0168-8278(00)80122-1. View

15.

Zhang C, Cai Y, Adachi M, Oshiro S, Aso T, Kaufman R . Homocysteine induces programmed cell death in human vascular endothelial cells through activation of the unfolded protein response. J Biol Chem. 2001; 276(38):35867-74. DOI: 10.1074/jbc.M100747200. View

16.

Zerbini L, Libermann T . Life and death in cancer. GADD45 alpha and gamma are critical regulators of NF-kappaB mediated escape from programmed cell death. Cell Cycle. 2004; 4(1):18-20. DOI: 10.4161/cc.4.1.1363. View

17.

McCullough K, Martindale J, Klotz L, Aw T, Holbrook N . Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bcl2 and perturbing the cellular redox state. Mol Cell Biol. 2001; 21(4):1249-59. PMC: 99578. DOI: 10.1128/MCB.21.4.1249-1259.2001. View

18.

Neuman M . Apoptosis in diseases of the liver. Crit Rev Clin Lab Sci. 2001; 38(2):109-66. DOI: 10.1080/20014091084182. View

19.

Werstuck G, Lentz S, Dayal S, Hossain G, Sood S, Shi Y . Homocysteine-induced endoplasmic reticulum stress causes dysregulation of the cholesterol and triglyceride biosynthetic pathways. J Clin Invest. 2001; 107(10):1263-73. PMC: 209295. DOI: 10.1172/JCI11596. View

20.

Deaciuc I, DSouza N, de Villiers W, Burikhanov R, Sarphie T, Hill D . Inhibition of caspases in vivo protects the rat liver against alcohol-induced sensitization to bacterial lipopolysaccharide. Alcohol Clin Exp Res. 2001; 25(6):935-43. View