Metformin exerts anti‑inflammatory effects on mouse colon smooth muscle cells in vitro

Al‑Dwairi,Ahmed; Alqudah,Mohammad; Al‑Shboul,Othman; Alfaqih,Mahmoud; Alomari,Dana

doi:10.3892/etm.2018.6222

Metformin exerts anti‑inflammatory effects on mouse colon smooth muscle cells in vitro

Authors:
- Ahmed Al‑Dwairi
- Mohammad Alqudah
- Othman Al‑Shboul
- Mahmoud Alfaqih
- Dana Alomari
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Affiliations: Department of Physiology and Biochemistry, Faculty of Medicine, Jordan University of Science and Technology, Irbid 22110, Jordan, Faculty of Medicine, Jordan University of Science and Technology, Irbid 22110, Jordan
Published online on: May 25, 2018 https://doi.org/10.3892/etm.2018.6222
Pages: 985-992

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Abstract

Inflammatory bowel disease (IBD) is a chronic incurable condition characterized by relapsing inflammation of the gut. Intestinal smooth muscle cells (SMCs) are affected structurally and functionally during IBD due to excessive production of different inflammatory mediators. Metformin is a widely used antidiabetic agent known to exert several anti‑inflammatory effects in different tissues independently from its hypoglycemic effect. The aim of the present study was to investigate the effect of metformin on expression and secretion of different cytokines and chemokines from mouse colon SMCs (CSMCs) following induction of inflammation with lipopolysaccharide (LPS) in vitro. CSMCs from male BALB/c mice were isolated and cultured in Dulbecco's modified Eagle's medium and treated with LPS (1 µg/ml) and 0, 5, 10 or 20 mM metformin for 24 h. Expression and secretion of tumor necrosis factor‑α (TNF‑α), interleukin‑1α (IL‑1α), macrophage colony stimulating factor (M‑CSF), T cell activation gene‑3 (TCA‑3) and stromal cell‑derived factor‑1 (SDF‑1) was evaluated by ELISA. LPS‑treated CSMCs demonstrated significantly increased expression of TNF‑α, IL‑1α, M‑CSF, TCA‑3 and SDF‑1 when compared with the control group (P<0.05). Co‑treatment with metformin (5 and 10 mM) significantly reduced their expression by ~20‑40% when compared with LPS treatment alone (P<0.05). Furthermore, secretion of TNF‑α, IL‑1α, M‑CSF and TCA‑3 into the conditioned media was significantly decreased by metformin (5 and 10 mM; P<0.05). In addition, metformin decreased levels of LPS‑induced nuclear factor‑κB phosphorylation. These data suggest that metformin may provide beneficial anti‑inflammatory effects on CSMCs and it may be utilized as an adjunct therapy for patients suffering from IBD.

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1	Snape WJ Jr, Williams R and Hyman PE: Defect in colonic smooth muscle contraction in patients with ulcerative colitis. Am J Physiol. 261:G987–G991. 1991.PubMed/NCBI
2	Severi C, Sferra R, Scirocco A, Vetuschi A, Pallotta N, Pronio A, Caronna R, Di Rocco G, Gaudio E, Corazziari E and Onori P: Contribution of intestinal smooth muscle to Crohn's disease fibrogenesis. Eur J Histochem. 58:24572014. View Article : Google Scholar : PubMed/NCBI
3	Vermillion DL, Huizinga JD, Riddell RH and Collins SM: Altered small intestinal smooth muscle function in Crohn's disease. Gastroenterology. 104:1692–1699. 1993. View Article : Google Scholar : PubMed/NCBI
4	Vrees MD, Pricolo VE, Potenti FM and Cao W: Abnormal motility in patients with ulcerative colitis: The role of inflammatory cytokines. Arch Surg. 137:439–446. 2002. View Article : Google Scholar : PubMed/NCBI
5	Shea-Donohue T, Notari L, Sun R and Zhao A: Mechanisms of smooth muscle responses to inflammation. Neurogastroenterol Motil. 24:802–811. 2012. View Article : Google Scholar : PubMed/NCBI
6	Shi XZ and Sarna SK: Transcriptional regulation of inflammatory mediators secreted by human colonic circular smooth muscle cells. Am J Physiol Gastrointest Liver Physiol. 289:G274–G284. 2005. View Article : Google Scholar : PubMed/NCBI
7	Salinthone S, Singer CA and Gerthoffer WT: Inflammatory gene expression by human colonic smooth muscle cells. Am J Physiol Gastrointest Liver Physiol. 287:G627–G637. 2004. View Article : Google Scholar : PubMed/NCBI
8	Abraham C and Cho JH: Inflammatory bowel disease. N Engl J Med. 361:2066–2078. 2009. View Article : Google Scholar : PubMed/NCBI
9	Bähler C, Schoepfer AM, Vavricka SR, Brüngger B and Reich O: Chronic comorbidities associated with inflammatory bowel disease: Prevalence and impact on healthcare costs in Switzerland. Eur J Gastroenterol Hepatol. 29:916–925. 2017. View Article : Google Scholar : PubMed/NCBI
10	Tamez-Pérez HE, Quintanilla-Flores DL, Rodríguez-Gutiérrez R, González-González JG and Tamez-Peña AL: Steroid hyperglycemia: Prevalence, early detection and therapeutic recommendations: A narrative review. World J Diabetes. 6:1073–1081. 2015. View Article : Google Scholar : PubMed/NCBI
11	Fakhoury M, Negrulj R, Mooranian A and Al-Salami H: Inflammatory bowel disease: Clinical aspects and treatments. J Inflamm Res. 7:113–120. 2014. View Article : Google Scholar : PubMed/NCBI
12	Pernicova I and Korbonits M: Metformin-mode of action and clinical implications for diabetes and cancer. Nat Rev Endocrinol. 10:143–156. 2014. View Article : Google Scholar : PubMed/NCBI
13	Cameron AR, Morrison VL, Levin D, Mohan M, Forteath C, Beall C, McNeilly AD, Balfour DJ, Savinko T, Wong AK, et al: Anti-inflammatory effects of metformin irrespective of diabetes status. Circ Res. 119:652–665. 2016. View Article : Google Scholar : PubMed/NCBI
14	Hirsch HA, Iliopoulos D and Struhl K: Metformin inhibits the inflammatory response associated with cellular transformation and cancer stem cell growth. Proc Natl Acad Sci USA. 110:972–977. 2013. View Article : Google Scholar : PubMed/NCBI
15	Kim SA and Choi HC: Metformin inhibits inflammatory response via AMPK-PTEN pathway in vascular smooth muscle cells. Biochem Biophys Res Commun. 425:866–872. 2012. View Article : Google Scholar : PubMed/NCBI
16	Kita Y, Takamura T, Misu H, Ota T, Kurita S, Takeshita Y, Uno M, Matsuzawa-Nagata N, Kato K, Ando H, et al: Metformin prevents and reverses inflammation in a non-diabetic mouse model of nonalcoholic steatohepatitis. PLoS One. 7:e430562012. View Article : Google Scholar : PubMed/NCBI
17	Cao X, Li H, Tao H, Wu N, Yu L, Zhang D, Lu X, Zhu J, Lu Z and Zhu Q: Metformin inhibits vascular calcification in female rat aortic smooth muscle cells via the AMPK-eNOS-NO pathway. Endocrinology. 154:3680–3689. 2013. View Article : Google Scholar : PubMed/NCBI
18	Lee SY, Lee SH, Yang EJ, Kim EK, Kim JK, Shin DY and Cho ML: Metformin ameliorates inflammatory bowel disease by suppression of the STAT3 signaling pathway and regulation of the between Th17/Treg balance. PLoS One. 10:e01358582015. View Article : Google Scholar : PubMed/NCBI
19	Isoda K, Young JL, Zirlik A, MacFarlane LA, Tsuboi N, Gerdes N, Schönbeck U and Libby P: Metformin inhibits proinflammatory responses and nuclear factor-kappaB in human vascular wall cells. Arterioscler Thromb Vasc Biol. 26:611–617. 2006. View Article : Google Scholar : PubMed/NCBI
20	Al-Dwairi A, Alqudah TE, Al-Shboul O, Alqudah M, Mustafa AG and Alfaqih MA: Glucagon-like peptide-1 exerts anti-inflammatory effects on mouse colon smooth muscle cells through the cyclic adenosine monophosphate/nuclear factor-κB pathway in vitro. J Inflamm Res. 11:95–109. 2018. View Article : Google Scholar : PubMed/NCBI
21	Berkes J, Viswanathan VK, Savkovic SD and Hecht G: Intestinal epithelial responses to enteric pathogens: Effects on the tight junction barrier, ion transport, and inflammation. Gut. 52:439–451. 2003. View Article : Google Scholar : PubMed/NCBI
22	Brynskov J, Foegh P, Pedersen G, Ellervik C, Kirkegaard T, Bingham A and Saermark T: Tumour necrosis factor alpha converting enzyme (TACE) activity in the colonic mucosa of patients with inflammatory bowel disease. Gut. 51:37–43. 2002. View Article : Google Scholar : PubMed/NCBI
23	Neurath MF: Cytokines in inflammatory bowel disease. Nat Rev Immunol. 14:329–342. 2014. View Article : Google Scholar : PubMed/NCBI
24	Rizzo G, Pugliese D, Armuzzi A and Coco C: Anti-TNF alpha in the treatment of ulcerative colitis: A valid approach for organ-sparing or an expensive option to delay surgery? World J Gastroenterol. 20:4839–4845. 2014. View Article : Google Scholar : PubMed/NCBI
25	Lv R, Qiao W, Wu Z, Wang Y, Dai S, Liu Q and Zheng X: Tumor necrosis factor alpha blocking agents as treatment for ulcerative colitis intolerant or refractory to conventional medical therapy: A meta-analysis. PLoS One. 9:e866922014. View Article : Google Scholar : PubMed/NCBI
26	Scarpa M, Kessler S, Sadler T, West G, Homer C, McDonald C, de la Motte C, Fiocchi C and Stylianou E: The epithelial danger signal IL-1α is a potent activator of fibroblasts and reactivator of intestinal inflammation. Am J Pathol. 185:1624–1637. 2015. View Article : Google Scholar : PubMed/NCBI
27	Papadakis KA and Targan SR: Role of cytokines in the pathogenesis of inflammatory bowel disease. Annu Rev Med. 51:289–298. 2000. View Article : Google Scholar : PubMed/NCBI
28	Dionne S, D'Agata DD, Hiscott J, Vanounou T and Seidman EG: Colonic explant production of IL-1 and its receptor antagonist is imbalanced in inflammatory bowel disease (IBD). Clin Exp Immunol. 112:435–442. 1998. View Article : Google Scholar : PubMed/NCBI
29	Cominelli F and Pizarro TT: Interleukin-1 and interleukin-1 receptor antagonist in inflammatory bowel disease. Aliment Pharmacol Ther. 10:49–53. 1996. View Article : Google Scholar : PubMed/NCBI
30	Ghia JE, Galeazzi F, Ford DC, Hogaboam CM, Vallance BA and Collins S: Role of M-CSF-dependent macrophages in colitis is driven by the nature of the inflammatory stimulus. Am J Physiol Gastrointest Liver Physiol. 294:G770–G777. 2008. View Article : Google Scholar : PubMed/NCBI
31	Marshall D, Cameron J, Lightwood D and Lawson AD: Blockade of colony stimulating factor-1 (CSF-I) leads to inhibition of DSS-induced colitis. Inflamm Bowel Dis. 13:219–224. 2007. View Article : Google Scholar : PubMed/NCBI
32	Franzè E, Marafini I, de Simone V, Monteleone I, Caprioli F, Colantoni A, Ortenzi A, Crescenzi F, Izzo R, Sica G, et al: Interleukin-34 induces Cc-chemokine ligand 20 in gut epithelial cells. J Crohns Colitis. 10:87–94. 2016. View Article : Google Scholar : PubMed/NCBI
33	Werner L, Guzner-Gur H and Dotan I: Involvement of CXCR4/CXCR7/CXCL12 interactions in inflammatory bowel disease. Theranostics. 3:40–46. 2013. View Article : Google Scholar : PubMed/NCBI
34	Danese S and Gasbarrini A: Chemokines in inflammatory bowel disease. J Clin Pathol. 58:1025–1027. 2005. View Article : Google Scholar : PubMed/NCBI
35	Mazzucchelli L, Hauser C, Zgraggen K, Zgraggen K, Wagner HE, Hess MW, Laissue JA and Mueller C: Differential in situ expression of the genes encoding the chemokines MCP-1 and RANTES in human inflammatory bowel disease. J Pathol. 178:201–206. 1996. View Article : Google Scholar : PubMed/NCBI
36	Luo Y and Dorf ME: Beta-chemokine TCA3 binds to mesangial cells and induces adhesion, chemotaxis, and proliferation. J Immunol. 156:742–748. 1996.PubMed/NCBI
37	Scheerens H, Hessel E, de Waal-Malefyt R, Leach MW and Rennick D: Characterization of chemokines and chemokine receptors in two murine models of inflammatory bowel disease: IL-10-/-mice and Rag-2-/-mice reconstituted with CD4+ CD45RBhigh T cells. Eur J Immunol. 31:1465–1474. 2001. View Article : Google Scholar : PubMed/NCBI
38	Xia XM, Wang FY, Zhou J, Hu KF, Li SW and Zou BB: CXCR4 antagonist AMD3100 modulates claudin expression and intestinal barrier function in experimental colitis. PLoS One. 6:e272822011. View Article : Google Scholar : PubMed/NCBI
39	Koh SJ, Kim JM, Kim IK, Ko SH and Kim JS: Anti-inflammatory mechanism of metformin and its effects in intestinal inflammation and colitis-associated colon cancer. J Gastroenterol Hepatol. 29:502–510. 2014. View Article : Google Scholar : PubMed/NCBI
40	Kothari V, Galdo JA and Mathews ST: Hypoglycemic agents and potential anti-inflammatory activity. J Inflamm Res. 9:27–38. 2016.PubMed/NCBI
41	Salminen A, Hyttinen JM and Kaarniranta K: AMP-activated protein kinase inhibits NF-κB signaling and inflammation: Impact on healthspan and lifespan. J Mol Med (Berl). 89:667–676. 2011. View Article : Google Scholar : PubMed/NCBI
42	He C, Li H, Viollet B, Zou MH and Xie Z: AMPK suppresses vascular inflammation in vivo by inhibiting signal transducer and activator of transcription-1. Diabetes. 64:4285–4297. 2015. View Article : Google Scholar : PubMed/NCBI
43	Atreya I, Atreya R and Neurath MF: NF-kappaB in inflammatory bowel disease. J Intern Med. 263:591–596. 2008. View Article : Google Scholar : PubMed/NCBI
44	Nair DG, Miller KG, Lourenssen SR and Blennerhassett MG: Inflammatory cytokines promote growth of intestinal smooth muscle cells by induced expression of PDGF-Rβ. J Cell Mol Med. 18:444–454. 2014. View Article : Google Scholar : PubMed/NCBI
45	Guan Q and Zhang J: Recent advances: The imbalance of cytokines in the pathogenesis of inflammatory bowel disease. Mediators Inflamm. 2017:48102582017. View Article : Google Scholar : PubMed/NCBI