Chymotrypsin activity signals to intestinal epithelium by protease-activated receptor-dependent mechanisms.

Autor: Guignard S; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Saifeddine M; Department of Physiology and Pharmacology, University of Calgary Cumming School of Medicine, Calgary, Alberta, Canada.; Department of Medicine, University of Calgary Cumming School of Medicine, Calgary, Alberta, Canada., Mihara K; Department of Physiology and Pharmacology, University of Calgary Cumming School of Medicine, Calgary, Alberta, Canada.; Department of Medicine, University of Calgary Cumming School of Medicine, Calgary, Alberta, Canada., Motahhary M; Department of Physiology and Pharmacology, University of Calgary Cumming School of Medicine, Calgary, Alberta, Canada.; Department of Medicine, University of Calgary Cumming School of Medicine, Calgary, Alberta, Canada., Savignac M; Toulouse Institute for Infectious and Inflammatory Diseases (Infinity) INSERM UMR1291-Centre National de la Recherche Scientifique UMR5051, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Guiraud L; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Sagnat D; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Sebbag M; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Khou S; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Rolland C; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Edir A; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Bournet B; Department of Gastroenterology, Toulouse University Hospital, Toulouse, France., Buscail L; Department of Gastroenterology, Toulouse University Hospital, Toulouse, France., Buscail E; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France.; Department of Digestive Surgery, Toulouse University Hospital, Toulouse, France., Alric L; Department of Internal Medicine and Digestive Diseases, Rangueil, Toulouse III University Hospital, University of Toulouse, Toulouse, France., Camare C; Department of Clinical Biochemistry, Toulouse University Hospital, Toulouse, France.; University of Toulouse, UMR1297, INSERM/Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Ambli M; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Vergnolle N; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France.; Department of Physiology and Pharmacology, University of Calgary Cumming School of Medicine, Calgary, Alberta, Canada., Hollenberg MD; Department of Physiology and Pharmacology, University of Calgary Cumming School of Medicine, Calgary, Alberta, Canada.; Department of Medicine, University of Calgary Cumming School of Medicine, Calgary, Alberta, Canada., Deraison C; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France., Bonnart C; IRSD, University of Toulouse, INSERM, INRAE, ENVT, Université Toulouse III-Paul Sabatier (UPS), Toulouse, France.
Jazyk: angličtina
Zdroj: British journal of pharmacology [Br J Pharmacol] 2024 Aug; Vol. 181 (16), pp. 2725-2749. Date of Electronic Publication: 2024 Apr 18.
DOI: 10.1111/bph.16341
Abstrakt: Background and Purpose: Chymotrypsin is a pancreatic protease secreted into the lumen of the small intestine to digest food proteins. We hypothesized that chymotrypsin activity may be found close to epithelial cells and that chymotrypsin signals to them via protease-activated receptors (PARs). We deciphered molecular pharmacological mechanisms and gene expression regulation for chymotrypsin signalling in intestinal epithelial cells.
Experimental Approach: The presence and activity of chymotrypsin were evaluated by Western blot and enzymatic activity tests in the luminal and mucosal compartments of murine and human gut samples. The ability of chymotrypsin to cleave the extracellular domain of PAR1 or PAR2 was assessed using cell lines expressing N-terminally tagged receptors. The cleavage site of chymotrypsin on PAR1 and PAR2 was determined by HPLC-MS analysis. The chymotrypsin signalling mechanism was investigated in CMT93 intestinal epithelial cells by calcium mobilization assays and Western blot analyses of (ERK1/2) phosphorylation. The transcriptional consequences of chymotrypsin signalling were analysed on colonic organoids.
Key Results: We found that chymotrypsin was present and active in the vicinity of the colonic epithelium. Molecular pharmacological studies have shown that chymotrypsin cleaves both PAR1 and PAR2 receptors. Chymotrypsin activated calcium and ERK1/2 signalling pathways through PAR2, and this pathway promoted interleukin-10 (IL-10) up-regulation in colonic organoids. In contrast, chymotrypsin disarmed PAR1, preventing further activation by its canonical agonist, thrombin.
Conclusion and Implications: Our results highlight the ability of chymotrypsin to signal to intestinal epithelial cells via PARs, which may have important physiological consequences in gut homeostasis.
(© 2024 The Authors. British Journal of Pharmacology published by John Wiley & Sons Ltd on behalf of British Pharmacological Society.)
Databáze: MEDLINE
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