EFFECT OF INSULIN ON LPS-INDUCED INFLAMMATORY MARKERS IN MOUSE COLON SMOOTH MUSCLE CELLS IN VITRO
Â Objective: The aim of the research is to determine the effect of supraphysiological doses of insulin on mouse colon smooth muscle cells (CSMCs) expression and secretion of pro-inflammatory cytokines interleukin 1 alpha (IL-1Î±) and tumor necrosis factor alpha (TNF-Î±) in vitro.
Methods: Freshly isolated CSMCs from BALBc mice were cultured in Dulbeccoâ€™s Modified Eagle Medium and treated with various doses of insulin (0, 1, 5, 10, and 20 nM) for 48 h, with/without lipopolysaccharides (LPS; 1 ug/mL) to induce inflammation. The levels of IL-1Î± and TNF-Î± in the cell homogenates and conditioned media were measured using ELISA.
Results: Insulin alone (1, 5, 10, and 20 nM) did not elicit a significant change in the expression or secretion of IL-1Î± or TNF-Î± form CSMCs; however, insulin (10 and 20 nM) significantly (p<0.05) increased the expression and secretion (~1.3â€“1.6-fold) of both IL-1Î± and TNF-Î± from CSMCs in the presence of inflammatory stimulus LPS when compared to LPS alone.
Conclusion: This study highlights the role of hyperinsulinemia on CSMC inflammation and its potential role in the pathogenesis of inflammatory bowel disease (IBD) during obesity. Measures that prevent obesity may protect against the development of IBD since the worldwide incidence of both obesity and IBD is increasing in a parallel fashion.
2. Singh S, Dulai PS, Zarrinpar A, Ramamoorthy S, Sandborn WJ. Obesity in IBD: Epidemiology, pathogenesis, disease course and treatment outcomes. Nat Rev Gastroenterol Hepatol 2017;14:110-21.
3. Bregenzer N, Hartmann A, Strauch U, SchÃ¶lmerich J, Andus T, Bollheimer LC, et al. Increased insulin resistance and beta cell activity in patients with Crohnâ€™s disease. Inflamm Bowel Dis 2006;12:53-6.
4. Nerkar D, Mukherjee A, Mehta BK, Banerjee S. Metabolic syndrome associated complications. Int J Pharm Pharm Sci 2015;4:22-5.
5. Bollapragada MK, Shantaram M, Kumar RS. Obesity: Development, epidemiology, factors affecting, quantity, health hazards, management and natural treatment - A review. Int J Pharm Pharm Sci 2017;9:12 26.
6. Harper JW, Zisman TL. Interaction of obesity and inflammatory bowel disease. World J Gastroenterol 2016;22:7868-81.
7. Baig MM, Fatima S, Fatima M, Siddiqui S, Ali SA, Ilyas MN. Prescription pattern and cost of illness (COI) of inflammatory bowel disease (IBD) in a tertiary care hospital. Int J Pharm Pharm Sci 2016;9:44-7.
8. Nascimento AT, Rocha R, Coqueiro FG, Santana GO, Lyra AC. Does obesity complicate inflammatory bowel diseases? J Crohns Colitis 2012;6:1041.
9. Seminerio JL, Koutroubakis IE, Ramos-Rivers C, Hashash JG, Dudekula A, Regueiro M, et al. Impact of obesity on the management and clinical course of patients with inflammatory bowel disease. Inflamm Bowel Dis 2015;21:2857-63.
10. Long MD, Crandall WV, Leibowitz IH, Duffy L, del Rosario F, Kim SC, et al. Prevalence and epidemiology of overweight and obesity in children with inflammatory bowel disease. Inflamm Bowel Dis 2011;17:2162-8.
11. Yorulmaz E, Adali G, Yorulmaz H, Ulasoglu C, Tasan G, Tuncer I, et al. Metabolic syndrome frequency in inflammatory bowel diseases. Saudi J Gastroenterol 2011;17:376-82.
12. Heymsfield SB, Wadden TA. Mechanisms, pathophysiology, and management of obesity. N Engl J Med 2017;376:254-66.
13. Iwasaki Y, Nishiyama M, Taguchi T, Asai M, Yoshida M, Kambayashi M, et al. Insulin exhibits short-term anti-inflammatory but long-term proinflammatory effects in vitro. Mol Cell Endocrinol 2009;298:25-32.
14. Perkins JM, Joy NG, Tate DB, Davis SN. Acute effects of hyperinsulinemia and hyperglycemia on vascular inflammatory biomarkers and endothelial function in overweight and obese humans. Am J Physiol Endocrinol Metab 2015;309:E168-76.
15. Sun Q, Li J, Gao F. New insights into insulin: The anti-inflammatory effect and its clinical relevance. World J Diabetes 2014;5:89-96.
16. Brundage SI, Kirilcuk NN, Lam JC, Spain DA, Zautke NA. Insulin increases the release of proinflammatory mediators. J Trauma 2008;65:367-72.
17. Sekine O, Nishio Y, Egawa K, Nakamura T, Maegawa H, Kashiwagi A. Insulin activates CCAAT/enhancer binding proteins and proinflammatory gene expression through the phosphatidylinositol 3-kinase pathway in vascular smooth muscle cells. J Biol Chem 2002;277:36631-9.
18. Manowsky J, Camargo RG, Kipp AP, Henkel J, PÃ¼schel GP. Insulin induced cytokine production in macrophages causes insulin resistance in hepatocytes. Am J Physiol Endocrinol Metab 2016;310:E938-46.
19. Guan Q, Zhang J. Recent advances: The imbalance of cytokines in the pathogenesis of inflammatory bowel disease. Mediators Inflamm 2017;2017:4810258.
20. Neurath MF. Cytokines in inflammatory bowel disease. Nat Rev Immunol 2014;14:329-42.
21. Lv R, Qiao W, Wu Z, Wang Y, Dai S, Liu Q, et al. Tumor necrosis factor alpha blocking agents as treatment for ulcerative colitis intolerant or refractory to conventional medical therapy: A meta-analysis. PLoS One 2014;9:e86692.
22. Cominelli F, Pizarro TT. Interleukin-1 and interleukin-1 receptor antagonist in inflammatory bowel disease. Aliment Pharmacol Ther 1996;10 Suppl 2:49-53.
23. Cardani D, Dusio GF, Luchini P, Sciarabba M, Solimene U, Rumio C, et al. Oral administration of interleukin-10 and anti IL-1 antibody ameliorates experimental intestinal inflammation. Gastroenterology Res 2013;6:124-33.
24. Scirocco A, Matarrese P, Carabotti M, Ascione B, Malorni W, Severi C, et al. Cellular and molecular mechanisms of phenotypic switch in gastrointestinal smooth muscle. J Cell Physiol 2016;231:295 302.
25. Vermillion DL, Huizinga JD, Riddell RH, Collins SM. Altered small intestinal smooth muscle function in Crohnâ€™s disease. Gastroenterology 1993;104:1692-9.
26. Snape WJ Jr., Williams R, Hyman PE. Defect in colonic smooth muscle contraction in patients with ulcerative colitis. Am J Physiol 1991;261:G987-91.
27. Nair DG, Han TY, Lourenssen S, Blennerhassett MG. Proliferation modulates intestinal smooth muscle phenotype in vitro and in colitis in vivo. Am J Physiol Gastrointest Liver Physiol 2011;300:G903-13.
28. Nair DG, Miller KG, Lourenssen SR, Blennerhassett MG. Inflammatory cytokines promote growth of intestinal smooth muscle cells by induced expression of PDGF-rÎ². J Cell Mol Med 2014;18:444-54.
29. Abraham C, Cho JH. Inflammatory bowel disease. N Engl J Med 2009;361:2066-78.
30. Klein AV, Kiat H. The mechanisms underlying fructose-induced hypertension: A review. J Hypertens 2015;33:912-20.
31. Dandona P, Chaudhuri A, Mohanty P, Ghanim H. Anti-inflammatory effects of insulin. Curr Opin Clin Nutr Metab Care 2007;10:511-7.
32. Scarpa M, Kessler S, Sadler T, West G, Homer C, McDonald C, et al. The epithelial danger signal IL-1alpha is a potent activator of fibroblasts and reactivator of intestinal inflammation. Am J Pathol 2015;185:1624 37.
33. Papadakis KA, Targan SR. Role of cytokines in the pathogenesis of inflammatory bowel disease. Annu Rev Med 2000;51:289-98.
34. Dionne S, Dâ€™Agata DD, Hiscott J, Vanounou T, Seidman EG. Colonic explant production of IL-1 and its receptor antagonist is imbalanced in inflammatory bowel disease (IBD). Clin Exp Immunol 1998;112:435 42.
35. Brynskov J, Foegh P, Pedersen G, Ellervik C, Kirkegaard T, Bingham A, et al. Tumour necrosis factor Î± converting enzyme (TACE) activity in the colonic mucosa of patients with inflammatory bowel disease. Gut 2002;51:37-43.
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