Involvement of MAPK/NF-κB signaling in the activation of the cholinergic anti-inflammatory pathway in experimental colitis by chronic vagus nerve stimulation
Peng Sun, Kewen Zhou, Sheng Wang, Ping Li, Sijuan Chen, Guiping Lin, Yan Zhao, Tinghuai Wang, Peng Sun, Kewen Zhou, Sheng Wang, Ping Li, Sijuan Chen, Guiping Lin, Yan Zhao, Tinghuai Wang
Abstract
Background: Autonomic nervous system dysfunction is implicated in the etiopathogenesis of inflammatory bowel diseases (IBD). Therapies that increase cardiovagal activity, such as Mind-Body interventions, are currently confirmed to be effective in clinical trials in IBD. However, a poor understanding of pathophysiological mechanisms limits the popularization of therapies in clinical practice. The aim of the present study was to explore the mechanisms of these therapies against 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis in rats using a chronic vagus nerve stimulation model in vivo, as well as the lipopolysaccharide (LPS)-induced inflammatory response in human epithelial colorectal adenocarcinoma cells (Caco-2) by acetylcholine in vitro.
Methods and results: Colitis was induced in rats with rectal instillation of TNBS, and the effect of chronic VNS (0.25 mA, 20 Hz, 500 ms) on colonic inflammation was evaluated. Inflammatory responses were assessed by disease activity index (DAI), histological scores, myeloperoxidase (MPO) activity, inducible nitric oxide synthase (iNOS), TNF-α and IL-6 production. The expression of Mitogen-activated protein kinases (MAPK) family members, IκB-α, and nuclear NF-κB p65 were studied by immunoblotting. Heart rate variability (HRV) analysis was also applied to assess the sympathetic-vagal balance. DAI, histological scores, MPO activity, iNOS, TNF-α and IL-6 levels were significantly decreased by chronic VNS. Moreover, both VNS and acetylcholine reduced the phosphorylation of MAPKs and prevented the nuclear translocation of NF-κB p65. Methyllycaconitine (MLA) only reversed the inhibitory effect on p-ERK and intranuclear NF-κB p65 expression by ACh in vitro, no significant change was observed in the expression of p-p38 MAPK or p-JNK by MLA.
Conclusion: Vagal activity modification contributes to the beneficial effects of the cholinergic anti-inflammatory pathway in IBD-related inflamed colonic mucosa based on the activation of MAPKs and nuclear translocation of NF-κB. Our work may provide key pathophysiological mechanistic evidence for novel therapeutic strategies that increase the cardiovagal activity in IBD patients.
Conflict of interest statement
Competing Interests: The authors have declared that no competing interests exist.
Figures
References
- Sands BE (2007) Inflammatory bowel disease: past, present, and future. J Gastroenterol 42: 16–25. doi:. PubMed: .
- Nathan C (2002) Points of control in inflammation. Nature 420: 846–852. doi:. PubMed: .
- O’Shea JJ, Murray PJ (2008) Cytokine signaling modules in inflammatory responses. Immunity 28: 477–487. doi:. PubMed: .
- Nathan CF (1989) Respiratory burst in adherent human neutrophils: triggering by colony-stimulating factors CSF-GM and CSF-G. Blood 73: 301–306. PubMed: .
- Babior BM (2000) Phagocytes and oxidative stress. Am J Med 109: 33–44. doi:. PubMed: .
- Ley K, Laudanna C, Cybulsky MI, Nourshargh S (2007) Getting to the site of inflammation: the leukocyte adhesion cascade updated. Nat Rev Immunol 7: 678–689. doi:. PubMed: .
- Podolsky DK (2002) Inflammatory bowel disease. N Engl J Med 347: 417-429. doi:. PubMed: .
- Rutgeerts P, Vermeire S, Van Assche G (2009) Biological therapies for inflammatory bowel diseases. Gastroenterology 136: 1182-1197. doi:. PubMed: .
- Langmead L, Rampton DS (2006) Review article: complementary and alternative therapies for inflammatory bowel disease. Aliment Pharmacol Ther 23: 341–349. doi:. PubMed: .
- Petersen AM, Pedersen BK (2005) The anti-inflammatory effect of exercise. J Appl Physiol 98: 1154-1162. doi:. PubMed: .
- Xu XY, Gao J, Ling D, Wang TH (2007) Biofeedback treatment of prehypertension: Analyses of efficacy, heart rate variability and EEG approximate entropy. J Hum Hypertens 12: 973–975. PubMed: .
- Wang SZ, Li S, Xu XY, Lin GP, Shao L et al. (2010) Effect of slow abdominal breathing combined with biofeedback on blood pressure and heart rate variability in pre-hypertension. J Altern Complement Med 16: 1039-1045. doi:. PubMed: .
- Ollsson B, Sundkvist G, Lindgren S (2007) Subclinical sympathetic neuropathy appears early in the course of Crohn’s disease. BMC Gastroenterol 7: 1-6. doi:. PubMed: .
- Taylor CT, Keely SJ (2007) The autonomic nervous system and inflammatory bowel disease. Auton Neurosci 133: 104-114. doi:. PubMed: .
- Maunder RG, Greenberg GR (2012) Relationship between social support and autonomic function during a stress protocol in ulcerative colitis patients in remission. Inflamm Bowel Dis 18: 737-742. doi:. PubMed: .
- Pellissier S, Dantzer C, Canini F, Mathieu N, Bonaz B (2010) Psychological adjustment and autonomic disturbances in inflammatory bowel diseases and irritable bowel syndrome. Psychoneuroendocrinology 35: 653-662. doi:. PubMed: .
- Borovikova LV, Ivanova S, Zhang M, Yang H, Botchkina GI et al. (2000) Vagus nerve stimulation attenuates the systemic inflammatory response to endotoxin. Nature 405: 458–462. doi:. PubMed: .
- Tracey KJ (2002) The inflammatory reflex. Nature 420: 853–859. doi:. PubMed: .
- Tracey KJ (2007) Physiology and immunology of the cholinergic anti-inflammatory pathway. J Clin Invest 117: 289–296. doi:. PubMed: .
- Tracey KJ (2009) Reflex control of immunity. Nat Rev Immunol 9: 418–428. doi:. PubMed: .
- Rosas-Ballina M, Tracey KJ (2009) Cholinergic control of inflammation. J Intern Med 265: 663–679. doi:. PubMed: .
- de Jonge WJ, Ulloa L (2007) The alpha7 nicotinic acetylcholine receptor as a pharmacological target for inflammation. Br J Pharmacol 151:915-529.
- Ghia JE, Collins SM (2006) The vagus nerve: a tonic inhibitory influence associated with inflammatory bowel disease in a murine model. Gastroenterology 131: 1122-1130. doi:. PubMed: .
- Ghia JE, Collins SM (2007) The protective effect of the vagus nerve in a murine model of chronic relapsing colitis. Am J Physiol Gastrointest Liver Physiol 293: 711-718. doi:.
- Meregnani J, Bonaz B (2011) Anti-inflammatory effect of vagus nerve stimulation in a rat model of inflammatory bowel disease. Auton Neurosci 160: 82–89. doi:. PubMed: .
- Coskun M, Olsen J, Seidelin JB, Nielsen OH (2010) MAP kinases in inflammatory bowel disease. Clin Chim Acta 412: 513–520. PubMed: .
- Rosillo MA, de la Lastra CA (2012) Dietary supplementation of an ellagic acid-enriched pomegranate extract attenuates chronic colonic inflammation in rats. Pharmacol Res 66: 235–242. doi:. PubMed: .
- Atreya I, Atreya R, Neurath MF (2008) NF-kappaB in inflammatory bowel disease. J Intern Med 263: 591–596. doi:. PubMed: .
- Vallabhapurapu S, Karin M (2009) Regulation and function of NF-kappaB transcription factors in the immune system. Annu Rev Immunol 27: 693–733. doi:. PubMed: .
- Bonaz BL, Bernstein CN (2013) Brain-gut interactions in inflammatory bowel disease. Gastroenterology 144: 36-49. doi:. PubMed: .
- Nolan RP, Floras JS, Ahmed L, Harvey PJ, Hiscock N et al. (2012) Behavioural modification of cholinergic anti-inflammatory response to C-reactive protein in patients with hypertension. J Intern Med 272: 161-169. doi:. PubMed: .
- Tracey KJ (2007) Physiology and immunology of the cholinergic anti-inflammatory pathway. J Clin Invest 117: 289–296. doi:. PubMed: .
- Reyt S, Picq C, Sinniger V, Clarençon D, Bonaz B et al. (2010) Dynamic Causal Modelling and physiological confounds: a functional MRI study of vagus nerve stimulation. NeuroImage 52: 1456–1464. doi:. PubMed: .
- Morris GP, Beck PL, Herridge MS, Depew WT, Szewczuk MR et al. (1989) Hapten-induced model of chronic inflammation and ulceration in the rat colon. Gastrogenterology 96: 795-803. PubMed: .
- Brenna Ø, Furnes MW, Drozdov I, van Beelen Granlund A, Flatberg A et al. (2013) Relevance of TNBS-Colitis in Rats: A Methodological Study with Endoscopic, Historical and Transcripttomic Characterization and Correlation to IBD. PLOS ONE (in press).
- Rosillo MA, Sánchez-Hidalgo M, Cárdeno A, Aparicio-Soto M, Sánchez-Fidalgo S et al. (2012) Supplementation of an ellagic acid-enriched pomegranate extract attenuates chronic colonic inflammation in rats. Pharmacol Res 66: 235-242. doi:. PubMed: .
- Roncucci L, Mora E, Mariani F, Bursi S, Pezzi A et al. (2008) Myeloperoxidasepositive cell infiltration in colorectal carcinogenesis as indicator of colorectal cancer risk. Cancer Epidemiol Biomarkers Prev 17: 2291–2297. doi:. PubMed: .
- Van Der Zanden EP, Boeckxstaens GE, de Jonge WJ (2009) The vagus nerve as a modulator of intestinal inflammation. Neurogastroenterol Motil 21: 6-17. doi:. PubMed: .
- Huston JM, Ochani M, Rosas-Ballina M, Liao H, Ochani K et al. (2006) Splenectomy inactivates the cholinergic anti-inflammatory pathway during lethal endotoxemia and polymicrobial sepsis. J Exp Med 203: 1623-1628. doi:. PubMed: .
- Rosas-Ballina M, Ochani M, Parrish WR, Ochani K, Harris YT et al. (2008) Splenic nerve is required for cholinergic anti-inflammatory pathway control of TNF in endotoxemia. Proc Natl Acad Sci U S A 105: 11008-11013. doi:. PubMed: .
- Rosas-Ballina M, Olofsson PS, Tracey KJ (2011) Acetylcholine-synthesizing T cells relay neural signals in a vagus nerve circuit. Science 334: 98-101. doi:. PubMed: .
- Huang J, Wang Y, Jiang D, Zhou J, Huang X (2010) The sympathetic-vagal balance against endotoxemia. J Neural Transm 117: 729–735. doi:. PubMed: .
- Billman GE (2011) Heart rate variability–a historical perspective. Front Physiol 2: 86 PubMed: .
- Tak PP, Firestein GS (2001) NF-κB: a key role in inflammatory diseases. J Clin Invest 107: 8-10.
- Broom OJ, Widjaya B, Troelsen J, Olsen J, Nielsen OH (2009) Mitogen activated protein kinases: a role in inflammatory bowel disease? Clin Exp Immunol 158: 272–280. doi:. PubMed: .
- Yoshikawa H, Kurokawa M, Ozaki N, Nara K, Atou K et al. (2006) Nicotine inhibits the production of proinflammatory mediators in human monocytes by suppression of I-κB phosphorylation and nuclear factor-κB transcriptional activity through nicotinic acetylcholine receptor α7. Clin Exp Immunol 146: 116-123. doi:. PubMed: .
- Snoek SA, Verstege MI, van der Zanden EP, Deeks N, Bulmer DC et al. (2010) Selective alpha7 nicotinic acetylcholine receptor agonists worsen disease in experimental colitis. Br J Pharmacol 160: 322-333. doi:. PubMed: .
- Nishitani Y, Zhang L, Yoshida M, Azuma T, Kanazawa K et al. (2013) Intestinal Anti-Inflammatory Activity of Lentinan: Influence on IL-8 and TNFR1 Expression in Intestinal Epithelial Cells. PLOS ONE, 8: e62441. doi:. PubMed: . PubMed:
- Bevilacqua MA, Faniello MC, Iovine B, Russo T, Cimino F et al. (2002) Transcription factor NF-Y regulates differentiation of CaCo-2 cells. Arch Biochem Biophys 407: 39-44. doi:. PubMed: .
- Schroecksnadel S, Gostner J, Jenny M, Kurz K, Schennach H et al. (2013) Immunomodulatory effects in vitro of vitamin K antagonist acenocoumarol. Thromb Res, 131: e264–9. doi:. PubMed: . PubMed:
- Chen Z, Hagler J, Palombella VJ, Melandri F, Scherer D et al. (1995) Signal-induced site-specific phosphorylation targets I kappa B alpha to the ubiquitin-proteasome pathway. Genes Dev 9: 1586–1597. doi:. PubMed: .
- Li DL, Liu JJ, Liu BH, Hu H, Sun L et al. (2011) Acetylcholine inhibits hypoxia-induced tumor necrosis factor-α production via regulation of MAPKs phosphorylation in cardiomyocytes. J Cell Physiol 226: 1052-1059. doi:. PubMed: .
- Gubbins EJ, Gopalakrishnan M, Li J (2010). Alpha 7 nAChR-mediated activation of MAP kinase pathways in PC12 cells. Brain Res 1328:1-11.
- Orr-Urtreger A, Kedmi M, Rosner S, Karmeli F, Rachmilewitz D (2005) Increased severity of experimental colitis in alpha 5 nicotinic acetylcholine receptor subunit-deficient mice. Neuroreport 16: 1123-1127. doi:. PubMed: .
- Cucina A, Dinicola S, Coluccia P, Proietti S, D’Anselmi F et al. (2012) Nicotine stimulates proliferation and inhibits apoptosis in colon cancer cell lines through activation of survival pathways. J Surg Res 178: 233-241. doi:. PubMed: .
- Handforth A, Krahl SE (2001) Suppression of harmaline-induced tremor in rats by vagus nerve stimulation. Mov Disord 16: 84-88. doi:. PubMed: .
- Manta S, El Mansari M, Debonnel G, Blier P (2013) Electrophysiological and neurochemical effects of long-term vagus nerve stimulation on the rat monoaminergic systems. Int J Neuropsychopharmacol 16: 459-470. doi:. PubMed: .
- Morris GP, Beck PL, Herridge MS, Depew WT, Szewczuk MR et al. (1989) Hapten-induced model of chronic inflammation and ulceration in the rat colon. Gastroenterology 96: 795-803. PubMed: .
- Sackeim HA, Rush AJ, George MS, Marangell LB, Husain MM et al. (2001) Vagus nerve stimulation (VNS) for treatmentresistant depression: efficacy, side effects, and predictors of outcome. Neuropsychopharmacology 25: 713-728. doi:. PubMed: .
- Lo Giudice P, Careddu A, Magni G, Quagliata T, Pacifici L et al. (2002) Autonomic neuropathy in streptozotocin diabetic rats: effect of acetyl-L-carnitine. Diabetes Res Clin Pract 56: 173-180. doi:. PubMed: .
- Cooper HS, Murthy SN, Shah RS, Sedergran DJ (1999) Clinicopathologic study of dextran sulfate sodium experimental murine colitis. Lab Invest 69: 238–249. PubMed: .
- Tsune I, Ikejima K, Hirose M, Yoshikawa M, Enomoto N et al. (2003) Dietary glycine prevents chemical-induced experimental colitis in the rat. Gastroenterology 125: 775-785. doi:. PubMed: .
- Hahm KB, Im YH, Parks TW, Park SH, Markowitz S et al. (2001) Loss of transforming growth factor beta signalling in the intestine contributes to tissue injury in inflammatory bowel disease. Gut 49: 190-198. doi:. PubMed: .
- Bozkurt A, Cakir B, Ercan F, Yeğen BC (2003) Anti-inflammatory effects of leptin and cholecystokinin on acetic acid-induced colitis in rats: role of capsaicin-sensitive vagal afferent fibers. Regul Pept 116: 109-118. doi:. PubMed: .
- Ryoyama K, Nomura T, Nakamura S (1993) Inhibition of macrophage nitric-oxide production by arachidonate-cascade inhibitors. Cancer Immunol Immun 37: 385-391. doi:.
- Patil VK, David M (2007) Hepatotoxic potential of Malathion in the freshwater teleost, Cirrhinus mrigala (Hamilton). J Basic Clin Physiol Pharmacol 18: 307-314. PubMed: .
Source: PubMed