BCL2 Regulates Differentiation of Intestinal Fibroblasts

Autor: Céline Mamie, Bradford L. McRae, Gerhard Rogler, Pedro A. Ruiz, Stephen H. Clarke, Martin Hausmann, Bruce Weder
Přispěvatelé: University of Zurich, Hausmann, Martin
Rok vydání: 2017
Předmět:
0301 basic medicine
Cell Culture Techniques
610 Medicine & health
Piperazines
Nitrophenols
03 medical and health sciences
Hydroxyproline
chemistry.chemical_compound
Mice
0302 clinical medicine
Western blot
Fibrosis
hemic and lymphatic diseases
medicine
Immunology and Allergy
Animals
Humans
2715 Gastroenterology
Intestinal Mucosa
Aged
Sulfonamides
medicine.diagnostic_test
Chemistry
Biphenyl Compounds
Dextran Sulfate
Gastroenterology
Cell Differentiation
Fibroblasts
Middle Aged
medicine.disease
Molecular biology
3. Good health
Transplantation
Intestines
Mice
Inbred C57BL
10219 Clinic for Gastroenterology and Hepatology
030104 developmental biology
Real-time polymerase chain reaction
Proto-Oncogene Proteins c-bcl-2
Apoptosis
2723 Immunology and Allergy
030211 gastroenterology & hepatology
Female
Myofibroblast
Transforming growth factor
Zdroj: Web of Science
ISSN: 1536-4844
Popis: Background Fibrosis in patients with Crohn's disease (CD) results from an imbalance toward excessive fibrous tissue formation driven by fibroblasts. Activation of fibroblasts is linked to the B-cell lymphoma 2 (BCL2) family, which is involved in the induction of apoptosis. We investigated the impact of BCL2 repression on fibrogenesis. Methods The model of dextran sodium sulfate (DSS)-induced chronic colitis and the heterotopic transplantation model of fibrosis were used. Following the administration of the BCL2 antagonist (ABT-737, 50 mg/kg/d), collagen layer thickness and hydroxyproline (HYP) content were determined. Fibroblasts were stimulated with the BCL2 antagonist (0.01-100 µM). BCL2, alpha smooth muscle actin (αSMA), and collagen I (COL1A1) were determined by quantitative polymerase chain reaction (qPCR), immunofluorescence microscopy (IF), and western blot (WB). mRNA expression pattern was determined by next-generation sequencing (NGS). Results Collagen layer thickness was significantly decreased in both DSS-induced chronic colitis and the transplantation model of fibrosis upon BCL2 antagonist administration compared with vehicle. Decreased HYP content confirmed the preventive effects of the BCL2 antagonist on fibrosis. In vitro, a significant increase in PI+/annexin V+ human colonic fibroblasts was determined by fluorescence-activated cell sorting upon treatment with high-dose BCL2 antagonist; at a lower dose, αSMA, COL1A1, and TGF were decreased. NGS, IF, and qPCR revealed decreased expression and nuclear translocation of GATA6 and SOX9, known for reprogramming fibroblasts. Conclusion BCL2 antagonist administration partially prevented fibrogenesis in both fibrosis models. The BCL2 antagonist reduced the expression of TGFβ-induced factors involved in differentiation of myofibroblasts, and therefore might represent a potential treatment option against CD-associated fibrosis.
Databáze: OpenAIRE
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