Inhibition of RIPK3 Pathway Attenuates Intestinal Inflammation and Cell Death of Inflammatory Bowel Disease and Suppresses Necroptosis in Peripheral Mononuclear Cells of Ulcerative Colitis Patients
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Receptor-interacting serine/threonine-protein kinase (RIPK) 3 is a member of the TNF receptor-I signaling complex and mediates necroptosis, an inflammatory cell death. Ulcerative colitis (UC) is an excessive inflammatory disease caused by uncontrolled T cell activation. The current study is aimed to determine whether RIPK3 inhibitor attenuates UC development inhibiting inflammation and necroptosis using experimental colitis mice model. Dextran sulfate sodium-induced colitis mice were administered RIPK3 inhibitor (3 mg/ml) 3 times and their tissues were analyzed by immunohistochemistry. RIPK3, mixed lineage kinase domain-like (MLKL), phosphorylated MLKL, IL-17, and CD4 in colitis patient colon tissues were detected using confocal microscopy. Protein levels were measured using immunohistochemistry and ELISA. The differentiation of Th17 cells was evaluated using flow cytometry. The expression of proinflammatory cytokines and necroptosis in peripheral blood mononuclear cells from UC patients was decreased markedly by RIPK3 inhibitor treatment. We also observed that the injection of RIPK3 inhibitor improves colitis severity and protects intestinal destruction. RIPK3 inhibitor reduced necroptosis factors and proinflammatory cytokines in the colon and consequently protected colon devastation. The expression of inflammatory mediators in experimental colitis mice splenocytes was decreased significantly by RIPK3 inhibitor treatment. These results suggest that RIPK3 inhibitor ameliorates severity of experimental colitis and reduces inflammation through the inhibition of inflammatory response and necroptosis and support RIPK3-targeting substances for treatment of UC.
Abstract
Receptor-interacting serine/threonine-protein kinase (RIPK) 3 is a member of the TNF receptor-I signaling complex and mediates necroptosis, an inflammatory cell death. Ulcerative colitis (UC) is an excessive inflammatory disease caused by uncontrolled T cell activation. The current study is aimed to determine whether RIPK3 inhibitor attenuates UC development inhibiting inflammation and necroptosis using experimental colitis mice model. Dextran sulfate sodium-induced colitis mice were administered RIPK3 inhibitor (3 mg/ml) 3 times and their tissues were analyzed by immunohistochemistry. RIPK3, mixed lineage kinase domain-like (MLKL), phosphorylated MLKL, IL-17, and CD4 in colitis patient colon tissues were detected using confocal microscopy. Protein levels were measured using immunohistochemistry and ELISA. The differentiation of Th17 cells was evaluated using flow cytometry. The expression of proinflammatory cytokines and necroptosis in peripheral blood mononuclear cells from UC patients was decreased markedly by RIPK3 inhibitor treatment. We also observed that the injection of RIPK3 inhibitor improves colitis severity and protects intestinal destruction. RIPK3 inhibitor reduced necroptosis factors and proinflammatory cytokines in the colon and consequently protected colon devastation. The expression of inflammatory mediators in experimental colitis mice splenocytes was decreased significantly by RIPK3 inhibitor treatment. These results suggest that RIPK3 inhibitor ameliorates severity of experimental colitis and reduces inflammation through the inhibition of inflammatory response and necroptosis and support RIPK3-targeting substances for treatment of UC.
참고문헌 (25)
- Galluzzi L, "Essential versus accessory aspects of cell death : recommendations of the NCCD 2015", Cell Death Differ, 22, pp.58~73 (2015)
- Pasparakis M, "Necroptosis and its role in inflammation", Nature, 517, pp.311~320 (2015)
- Vandenabeele P, "Molecular mechanisms of necroptosis : an ordered cellular explosion", Nat Rev Mol Cell Biol, 11, pp.700~714 (2010)
- Linkermann A, "Necroptosis", N Engl J Med, 370, pp.455~465 (2014)
- Vanden Berghe T, "Regulated necrosis : the expanding network of non-apoptotic cell death pathways", Nat Rev Mol Cell Biol, 15, pp.135~147 (2014)
- Ford AC, "Ulcerative colitis", BMJ, 346, pp.f432~ (2013)
- Baumgart DC, "Inflammatory bowel disease : clinical aspects and established and evolving therapies", Lancet, 369, pp.1641~1657 (2007)
- Dignass A, "Second European evidence-based consensus on the diagnosis and management of ulcerative colitis part 1 : definitions and diagnosis", J Crohns Colitis, 6, pp.965~990 (2012)
- Orholm M, "Concordance of inflammatory bowel disease among Danish twins. Results of a nationwide study", Scand J Gastroenterol, 35, pp.1075~1081 (2000)
- Ko IK, "Targeting improves MSC treatment of inflammatory bowel disease", Mol Ther, 18, pp.1365~1372 (2010)
- Geerling BJ, "Diet as a risk factor for the development of ulcerative colitis", Am J Gastroenterol, 95, pp.1008~1013 (2000)
- Lees CW, "New IBD genetics : common pathways with other diseases", Gut, 60, pp.1739~1753 (2011)
- Pierdomenico M, "Necroptosis is active in children with inflammatory bowel disease and contributes to heighten intestinal inflammation", Am J Gastroenterol, 109, pp.279~287 (2014)
- Geboes K, "A reproducible grading scale for histological assessment of inflammation in ulcerative colitis", Gut, 47, pp.404~409 (2000)
- Ito R, "Interferon-gamma is causatively involved in experimental inflammatory bowel disease in mice", Clin Exp Immunol, 146, pp.330~338 (2006)
- Feng T, "Th17 cells induce colitis and promote Th1 cell responses through IL-17 induction of innate IL-12 and IL-23 production", J Immunol, 186, pp.6313~6318 (2011)
- Lee SY, "Metformin ameliorates inflammatory bowel disease by suppression of the STAT3 signaling pathway and regulation of the between Th17", Treg balance, 2015, pp.10, e0135858~
- Moriwaki K, "The necroptosis adaptor RIPK3 promotes injury-induced cytokine expression and tissue repair", Immunity, 41, pp.567~578 (2014)
- Newton K, "RIPK3 deficiency or catalytically inactive RIPK1 provides greater benefit than MLKL deficiency in mouse models of inflammation and tissue injury", Cell Death Differ, 23, pp.1565~1576 (2016)
- Lau A, "RIPK3-mediated necroptosis promotes donor kidney inflammatory injury and reduces allograft survival", Am J Transplant, 13, pp.2805~2818 (2013)
- Liu ZY, "Necrostatin-1 reduces intestinal inflammation and colitis-associated tumorigenesis in mice", Am J Cancer Res, 5, pp.3174~3185 (2015)
- Sartor RB, "Mechanisms of disease : pathogenesis of Crohn's disease and ulcerative colitis", Nat Clin Pract Gastroenterol Hepatol, 3, pp.390~407 (2006)
- Brasseit J, "CD4 T cells are required for both development and maintenance of disease in a new mouse model of reversible colitis", Mucosal Immunol, 9, pp.689~701 (2016)
- Lee SH, "Interferon-gamma regulates inflammatory cell death by targeting necroptosis in experimental autoimmune arthritis", Sci Rep, 7, pp.10133~ (2017)
- Miossec P, "Targeting IL-17 and TH17 cells in chronic inflammation", Nat Rev Drug Discov, 11, pp.763~776 (2012)