Activation of Myofibroblast TRPA1 by Steroids and Pirfenidone Ameliorates Fibrosis in Experimental Crohn's Disease
| Autor: | Daibo Kojima, Hidetoshi Takedatsu, Kunihiko Aoyagi, Kaori Koga, Lin Hai Kurahara, Yuwen Jian, Miki Onitsuka, Mayumi Doi, Yaopeng Hu, Ryuji Inoue, Keizo Hiraishi, Yoshitaka Fujihara |
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| Rok vydání: | 2018 |
| Předmět: |
RT-PCR
reverse-transcription polymerase chain reaction 0301 basic medicine TNBS trinitrobenzene sulfonic acid TRPA1 transient receptor potential ankyrin 1 PCR polymerase chain reaction Fibrosis TGF transforming growth factor Original Research Heat shock protein 47 TNF tumor necrosis factor biology Gastroenterology food and beverages Pirfenidone mRNA messenger RNA Pathophysiology HSP47 heat shock protein 47 AITC allyl isothiocyanate Knockout mouse InMyoFib intestinal myofibroblast cell line medicine.symptom Myofibroblast psychological phenomena and processes TRP transient receptor potential Crohn’s Disease medicine.drug sgRNA single-guide RNA Intestinal Fibrosis PBS phosphate-buffered saline Inflammation 03 medical and health sciences MT Masson trichrome CD Crohn’s disease medicine lcsh:RC799-869 KO knockout Hepatology business.industry medicine.disease WT wild-type Transient Receptor Potential Ankyrin 1 030104 developmental biology siRNA small interfering RNA Myocardin biology.protein Cancer research lcsh:Diseases of the digestive system. Gastroenterology α-SMA α smooth muscle actin EGTA ethylene glycol-bis(β-aminoethyl ether)-N N N′ N′-tetraacetic acid TRPC transient receptor potential canonical business |
| Zdroj: | Cellular and Molecular Gastroenterology and Hepatology, Vol 5, Iss 3, Pp 299-318 (2018) Cellular and Molecular Gastroenterology and Hepatology |
| ISSN: | 2352-345X |
| DOI: | 10.1016/j.jcmgh.2017.12.005 |
| Popis: | Background & Aims The transient receptor potential ankyrin 1 (TRPA1) channel is highly expressed in the intestinal lamina propria, but its contribution to gut physiology/pathophysiology is unclear. Here, we evaluated the function of myofibroblast TRPA1 channels in intestinal remodeling. Methods An intestinal myofibroblast cell line (InMyoFibs) was stimulated by transforming growth factor-β1 to induce in vitro fibrosis. Trpa1 knockout mice were generated using the Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system. A murine chronic colitis model was established by weekly intrarectal trinitrobenzene sulfonic acid (TNBS) administration. Samples from the intestines of Crohn’s disease (CD) patients were used for pathologic staining and quantitative analyses. Results In InMyoFibs, TRPA1 showed the highest expression among TRP family members. In TNBS chronic colitis model mice, the extents of inflammation and fibrotic changes were more prominent in TRPA1-/- knockout than in wild-type mice. One-week enema administration of prednisolone suppressed fibrotic lesions in wild-type mice, but not in TRPA1 knockout mice. Steroids and pirfenidone induced Ca2+ influx in InMyoFibs, which was antagonized by the selective TRPA1 channel blocker HC-030031. Steroids and pirfenidone counteracted transforming growth factor-β1–induced expression of heat shock protein 47, type 1 collagen, and α-smooth muscle actin, and reduced Smad-2 phosphorylation and myocardin expression in InMyoFibs. In stenotic intestinal regions of CD patients, TRPA1 expression was increased significantly. TRPA1/heat shock protein 47 double-positive cells accumulated in the stenotic intestinal regions of both CD patients and TNBS-treated mice. Conclusions TRPA1, in addition to its anti-inflammatory actions, may protect against intestinal fibrosis, thus being a novel therapeutic target for highly incurable inflammatory/fibrotic disorders. Graphical abstract |
| Databáze: | OpenAIRE |
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