Hepatocyte CREBH deficiency aggravates inflammatory liver injury following chemokine-dependent neutrophil infiltration through upregulation of NF-κB p65 in mice
- 62 Downloads
Fulminant hepatitis is a serious inflammatory condition of the liver characterized by massive necrosis of liver parenchyma following excessive immune cell infiltration into the liver, and possibly causing sudden hepatic failure and medical emergency. However, the underlying mechanisms are not fully understood. Here, we investigated the role of cyclic AMP-responsive element-binding protein, hepatocyte specific (CREBH) in concanavalin A (ConA)-driven hepatitis-evoked liver injury. C57BL/6J (WT) and Crebh knockout (KO) mice injected with ConA (7.5 or 25 mg/kg) and bone marrow (BM) chimeric mice, generated by injection of BM cells into sub-lethally irradiated recipients followed by ConA injection (22.5 or 27.5 mg/kg) 8 weeks later, were used for in vivo study. Primary mouse hepatocytes and HEK293T cells were used for a comparative in vitro study. Crebh KO mice are highly susceptible to ConA-induced liver injury and prone to death due to increased neutrophil infiltration driven by enhanced hepatic expression of neutrophil-attracting chemokines. Notably, BM chimera experiment demonstrated that Crebh-deficient hepatocytes have an enhanced ability of recruiting neutrophils to the liver, thereby promoting hepatotoxicity by ConA. Intriguingly, in vitro assays showed that p65, a subunit of NF-κB and common transcription factor for various chemokines, dependent transactivation was inhibited by CREBH. Furthermore, p65 expression was inversely correlated with CREBH level in ConA-treated mice liver and TNFα-stimulated primary mouse hepatocytes. This is the first demonstration that CREBH deficiency aggravates inflammatory liver injury following chemokine-dependent neutrophil infiltration via NF-κB p65 upregulation. CREBH is suggested to be a novel therapeutic target for treatment of fulminant hepatitis.
KeywordsCREBH Concanavalin A Inflammation Liver injury Neutrophil NF-κB
This work was supported by the KRIBB Research Initiative Program of the Republic of Korea, by the Development of Platform Technology for Innovative Medical Measurements Program (No. KRISS-2019-GP2019-0013) from the Korea Research Institute of Standards and Science and by the National Research Foundation of Korea (NRF) and the Korean government (MSIP) (NRF-2019R1C1C1005319).
JRN, JHK, YHK and CHL designed the study, and drafted the manuscript. JRN, JHK, SYN, IBL, YJS, JHC, YS and YHK performed experiments, and collected and analyzed data for the study. TGL, HSC and CHL contributed to critical revisions of the text.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
All animal experiments were approved by the Institutional Animal Care and Use Committee of the KRIBB.
- De Plaen IG, Han XB, Liu X, Hsueh W, Ghosh S, May MJ (2006) Lipopolysaccharide induces CXCL2/macrophage inflammatory protein-2 gene expression in enterocytes via NF-kappaB activation: independence from endogenous TNF-alpha and platelet-activating factor. Immunology 118:153–163PubMedCentralCrossRefGoogle Scholar
- Dominici S, Visvikis A, Pieri L, Paolicchi A, Valentini MA, Comporti M, Pompella A (2003) Redox modulation of NF-kappaB nuclear translocation and DNA binding in metastatic melanoma. The role of endogenous and gamma-glutamyl transferase-dependent oxidative stress. Tumori 89:426–433PubMedCrossRefPubMedCentralGoogle Scholar
- Marra F, Tacke F (2014) Roles for chemokines in liver disease. Gastroenterology 147(577–594):e571Google Scholar
- Misra J, Chanda D, Kim DK, Li T, Koo SH, Back SH, Chiang JY, Choi HS (2011) Curcumin differentially regulates endoplasmic reticulum stress through transcriptional corepressor SMILE (small heterodimer partner-interacting leucine zipper protein)-mediated inhibition of CREBH (cAMP responsive element-binding protein H). J Biol Chem 286:41972–41984PubMedPubMedCentralCrossRefGoogle Scholar
- Noh JR, Kim YH, Kim DK, Hwang JH, Kim KS, Choi DH, Lee SJ, Lee HG, Lee TG, Weng HL et al (2018a) Small heterodimer partner deficiency increases inflammatory liver injury through C–X–C motif chemokine ligand 2-driven neutrophil recruitment in mice. Toxicol Sci 163:254–264PubMedCrossRefPubMedCentralGoogle Scholar
- Penido C, Vieira-de-Abreu A, Bozza MT, Castro-Faria-Neto HC, Bozza PT (2003) Role of monocyte chemotactic protein-1/CC chemokine ligand 2 on gamma delta T lymphocyte trafficking during inflammation induced by lipopolysaccharide or Mycobacterium bovis bacille Calmette-Guerin. J Immunol 171:6788–6794PubMedCrossRefPubMedCentralGoogle Scholar
- Reichel CA, Rehberg M, Lerchenberger M, Berberich N, Bihari P, Khandoga AG, Zahler S, Krombach F (2009) Ccl2 and Ccl3 mediate neutrophil recruitment via induction of protein synthesis and generation of lipid mediators. Arterioscler Thromb Vasc Biol 29:1787–1793PubMedCrossRefPubMedCentralGoogle Scholar
- Rezzonico R, Imbert V, Chicheportiche R, Dayer JM (2001) Ligation of CD11b and CD11c beta(2) integrins by antibodies or soluble CD23 induces macrophage inflammatory protein 1alpha (MIP-1alpha) and MIP-1beta production in primary human monocytes through a pathway dependent on nuclear factor-kappaB. Blood 97:2932–2940PubMedCrossRefGoogle Scholar
- Zhang C, Wang G, Zheng Z, Maddipati KR, Zhang X, Dyson G, Williams P, Duncan SA, Kaufman RJ, Zhang K (2012) Endoplasmic reticulum-tethered transcription factor cAMP responsive element-binding protein, hepatocyte specific, regulates hepatic lipogenesis, fatty acid oxidation, and lipolysis upon metabolic stress in mice. Hepatology 55:1070–1082PubMedPubMedCentralCrossRefGoogle Scholar