Abstract
Liver fibrosis is the final common pathway for almost all causes of chronic liver injury. This chronic disease is characterized by excessive deposition of extracellular matrix components mainly due to transdifferentiation of quiescent hepatic stellate cell into myofibroblasts-like cells, which in turn is driven by cell death and inflammation. In the last few years, paracrine signaling through pannexin1 channels has emerged as a key player in the latter processes. The current study was set up to investigate the role of pannexin1 signaling in liver fibrosis. Wild-type and whole body pannexin1 knock-out mice were treated with carbon tetrachloride or subjected to bile duct ligation. Evaluation of the effects of pannexin1 deletion was based on a number of clinically relevant read-outs, including markers of liver damage, histopathological analysis, oxidative stress, inflammation and regenerative capacity. In parallel, to elucidate the molecular pathways affected by pannexin1 deletion as well as to mechanistically anchor the clinical observations, whole transcriptome analysis of liver tissue was performed. While pannexin1 knock-out mice treated with carbon tetrachloride displayed reduced collagen content, hepatic stellate cell activation, inflammation and hepatic regeneration, bile duct ligated counterparts showed increased hepatocellular injury and antioxidant enzyme activity with a predominant immune response. Gene expression profiling revealed a downregulation of fibrotic and immune responses in pannexin1 knock-out mice treated with carbon tetrachloride, whereas bile duct ligated pannexin1-deficient animals showed a pronounced inflammatory profile. This study shows for the first time an etiology-dependent role for pannexin1 signaling in experimental liver fibrosis.
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Abbreviations
- α-SMA:
-
Alpha smooth muscle actin
- ALT:
-
Alanine aminotransferase
- ALP:
-
Alkaline phosphatase
- ANOVA:
-
Analysis of variance
- ASC:
-
Apoptosis-associated speck-like protein containing a C-terminal caspase-recruitment domain
- AST:
-
Aspartate aminotransferase
- ATP:
-
Adenosine-5′-triphosphate
- BDL:
-
Bile duct ligation
- Casp1:
-
Caspase 1
- CCl4 :
-
Carbon tetrachloride
- Clec7a:
-
Dectin-1
- Col1a1:
-
Collagen type 1 alpha 1
- Cxcl14:
-
Chemokine ligand 14
- GPx:
-
Glutathione peroxidase
- GR:
-
Glutathione reductase
- HSCs:
-
Hepatic stellate cells
- IL:
-
Interleukin
- ip:
-
Intraperitoneally
- Lbp:
-
Lipopolysaccharide-binding protein
- Loxl2:
-
Lysyl oxidase-like 2
- Ly96:
-
Lymphocyte antigen 96
- MCP-1:
-
Monocyte chemoattractant protein 1
- MIP1γ:
-
Macrophage protein 1 gamma
- Nalp3:
-
NACHT, LRR, and pyrin domain-containing protein 3
- n :
-
Number or repeats
- NF-κB:
-
Nuclear factor kappa B
- Panx:
-
Pannexin
- p :
-
Probability
- RT-qPCR:
-
Reverse transcription quantitative real-time polymerase chain reaction
- S100a9:
-
S100 calcium-binding protein A9
- SEM:
-
Standard error of mean
- SOD:
-
Superoxide dismutase
- sTNF-RI/-RII:
-
Soluble TNF receptor 1/2
- TCA-3:
-
T-cell activation protein 3
- TECK:
-
Thymus-expressed chemokine
- TLR:
-
Toll-like receptor
- TNFα:
-
Tumor necrosis factor alpha
- WT:
-
Wild-type
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Acknowledgements
This work was supported by the grants of the “Fundação de Auxílio à Pesquisa do Estado de São Paulo” (FAPESP Grants 14/23890-4; 14/23887-3 and SPEC 13/50420-6), the European Research Council (ERC Starting Grant 335476), the Fund for Scientific Research-Flanders (FWO Grants G009514N and G010214N) and the University Hospital of the Vrije Universiteit Brussel-Belgium (“Willy Gepts Fonds” UZ-VUB). The authors thank Miss Tineke Vanhalewyn, Miss Dinja De Win and Mr Steven Branson for their dedicated technical assistance.
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204_2018_2255_MOESM1_ESM.pdf
Western blots from p-NF-κBp65 S536 protein expression in liver fibrosis. WT and Panx1-/- mice (n = 10-15) were administered 20% CCl4 ip at a gradually increased dose for 8 weeks or subjected to BDL for 20 days. Representative blot membranes expressing p-NF-κBp65 S536 proteins, total protein loading and β-actin proteins from a the CCl4 model and b the BDL model (PDF 230 KB)
204_2018_2255_MOESM3_ESM.docx
Gene modulation by the CCl4 treatment independent of background: comparison between WT CCl4 vs WT-oil and Panx1-/- CCl4 vs WT-oil (DOCX 25 KB)
204_2018_2255_MOESM5_ESM.docx
Gene modulation by the BDL procedure independent of background: comparison between WT BDL vs WT-sham and Panx1-/- BDL vs WT-sham (DOCX 45 KB)
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Crespo Yanguas, S., da Silva, T.C., Pereira, I.V.A. et al. Genetic ablation of pannexin1 counteracts liver fibrosis in a chemical, but not in a surgical mouse model. Arch Toxicol 92, 2607–2627 (2018). https://doi.org/10.1007/s00204-018-2255-3
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DOI: https://doi.org/10.1007/s00204-018-2255-3