Essential role of mast cells in the visceral hyperalgesia induced by T. spiralis infection and stress in rats

Chang-Qing Yang, Yan-Yu Wei, Chan-Juan Zhong, Li-Ping Duan, Chang-Qing Yang, Yan-Yu Wei, Chan-Juan Zhong, Li-Ping Duan

Abstract

Mast cells (MCs) deficient rats (Ws/Ws) were used to investigate the roles of MCs in visceral hyperalgesia. Ws/Ws and wild control (+/+) rats were exposed to T. spiralis or submitted to acute cold restraint stress (ACRS). Levels of proteinase-activated receptor 2 (PAR2) and nerve growth factor (NGF) were determined by immunoblots and RT-PCR analysis, and the putative signal pathways including phosphorylated extracellular-regulated kinase (pERK1/2) and transient receptor potential vanilloid receptor 1 (TRPV1) were further identified. Visceral hyperalgesia triggered by ACRS was observed only in +/+ rats. The increased expression of PAR2 and NGF was observed only in +/+ rats induced by T. spiralis and ACRS. The activation of pERK1/2 induced by ACRS occurred only in +/+ rats. However, a significant increase of TRPV1 induced by T. spiralis and ACRS was observed only in +/+ rats. The activation of PAR2 and NGF via both TRPV1 and pERK1/2 signal pathway is dependent on MCs in ACRS-induced visceral hyperalgesia rats.

Figures

Figure 1
Figure 1
The distension volumes needed to reach an abdominal withdrawal reflex (AWR) score of 3 were significantly lower in postinfection (PI) rats, rats induced by acute cold restraint stress (ACRS), and PI rats received ACRS procedures (PI + ACRS). Each group represents the mean ± SEM of 6 rats. **P < 0.01.
Figure 2
Figure 2
The increased PAR2 expression induced by intestinal infection (PI) and acute cold restraint stress (ACRS) in distal colon of +/+, but not Ws/Ws, rats. (a) Representative western blotting for PAR2 in extracts from colon tissue. (b) Quantitative analysis of PAR2 protein. Data was expressed as normalized density to β-actin. (c) Relative levels of PAR2 mRNA in colon tissue. Data was normalized to 18S ribosomal RNA and expressed using the 2−ΔΔCt method. *P < 0.05; **P < 0.01.
Figure 3
Figure 3
Relative levels of NGF mRNA were increased in the distal colon in +/+ but not Ws/Ws rats induced by intestinal infection (PI) and acute cold restraint stress (ACRS). Data was normalized to 18S ribosomal RNA and expressed using the 2−ΔΔCt method. *P < 0.05.
Figure 4
Figure 4
Role of ERK phosphorylation in +/+ and Ws/Ws rats induced by intestinal infection (PI) and acute cold restraint stress (ACRS). (a–f) Representative immunofluorescence images of pERK1/2 immunoreactivity- (IR-) positive neurons in L6S1 DRGs in +/+ rats (a–c) and Ws/Ws rats (d–f). Scale bar: 100 μm. (g) Quantification of pERK1/2 IR labelling intensity in L6S1 DRGs. (h) Representative western blot for phosphorylated ERK1/2 in L6S1 DRG extracts using a phospho-ERK specific Ab (pERK1/2, upper panel). Protein loading was confirmed by reprobing the membrane with ERK1/2 Ab (lower panel). (i) Quantitative analysis of phosphorylated ERK1/2 protein. Data was expressed as normalized density to ERK1/2. *P < 0.05; **P < 0.01.
Figure 5
Figure 5
The expression of TRPV1 in L6S1 DRG neurons in +/+ and Ws/Ws rats induced by intestinal infection (PI) and acute cold restraint stress (ACRS). (a–f) Representative immunofluorescence images of TRPV1 immunoreactivity (IR)-positive neurons in L6S1 DRGs in +/+ rats (a–c) and Ws/Ws rats (d–f). Scare bar: 100 μm. (g) Quantification of TRPV1 IR labelling intensity in L6S1 DRGs. (h) Representative western blotting for TRPV1 in extracts from L6S1 DRG extracts in +/+ and Ws/Ws rats. (i) Quantitative analysis of TRPV1 protein. Data was expressed as normalized density to β-actin. *P < 0.05; **P < 0.01.

References

    1. Mayer EA, Gebhart GF. Basic and clinical aspects of visceral hyperalgesia. Gastroenterology. 1994;107(1):271–293.
    1. Ritchie J. Pain from distension of the pelvic colon by inflating a balloon in the irritable colon syndrome. Gut. 1973;14(2):125–132.
    1. Guilarte M, Santos J, de Torres I, et al. Diarrhoea-predominant IBS patients show mast cell activation and hyperplasia in the jejunum. Gut. 2007;56(2):203–209.
    1. Klooker TK, Braak B, Koopman KE, et al. The mast cell stabiliser ketotifen decreases visceral hypersensitivity and improves intestinal symptoms in patients with irritable bowel syndrome. Gut. 2010;59(9):1213–1221.
    1. Chadwick VS, Chen W, Shu D, et al. Activation of the mucosal immune system in irritable bowel syndrome. Gastroenterology. 2002;122(7):1778–1783.
    1. Barbara G, Stanghellini V, De Giorgio R, et al. Activated mast cells in proximity to colonic nerves correlate with abdominal pain in irritable bowel syndrome. Gastroenterology. 2004;126(3):693–702.
    1. Reed DE, Barajas-Lopez C, Cottrell G, et al. Mast cell tryptase and proteinase-activated receptor 2 induce hyperexcitability of guinea-pig submucosal neurons. The Journal of Physiology. 2003;547(2):531–542.
    1. Delafoy L, Gelot A, Ardid D, et al. Interactive involvement of brain derived neurotrophic factor, nerve growth factor, and calcitonin gene related peptide in colonic hypersensitivity in the rat. Gut. 2006;55(7):940–945.
    1. Dai Y, Moriyama T, Higashi T, et al. Proteinase-activated receptor 2-mediated potentiation of transient receptor potential vanilloid subfamily 1 activity reveals a mechanism for proteinase-induced inflammatory pain. The Journal of Neuroscience. 2004;24(18):4293–4299.
    1. Gu Q, Lee LY. Effect of protease-activated receptor 2 activation on single TRPV1 channel activities in rat vagal pulmonary sensory neurons. Experimental Physiology. 2009;94(8):928–936.
    1. van den Wijngaard RM, Klooker TK, Welting O, et al. Essential role for TRPV1 in stress-induced (mast cell-dependent) colonic hypersensitivity in maternally separated rats. Neurogastroenterology and Motility. 2009;21(10):p. 1107-e94.
    1. Fukushima O, Nishimura S, Matsunami M, et al. Phosphorylation of ERK in the spinal dorsal horn following pancreatic pronociceptive stimuli with proteinase-activated receptor-2 agonists and hydrogen sulfide in rats: evidence for involvement of distinct mechanisms. Journal of Neuroscience Research. 2010;88(14):3198–3205.
    1. Stamboulian S, Choi JS, Ahn HS, et al. ERK1/2 mitogen-activated protein kinase phosphorylates sodium channel Nav1.7 and alters its gating properties. The Journal of Neuroscience. 2010;30(5):1637–1647.
    1. Barone FC, Deegan JF, Price WJ, Fowler PJ, Fondacaro JD, Ormsbee HS. Cold-restraint stress increases rat fecal pellet output and colonic transit. American Journal of Physiology. 1990;258(3, part 1):G329–G337.
    1. Leng YX, Wei YY, Chen H, Zhou SP, Yang YL, Duan LP. Alteration of cholinergic and peptidergic neurotransmitters in rat ileum induced by acute stress following transient intestinal infection is mast cell dependent. Chinese Medical Journal. 2010;123(2):227–233.
    1. Ford AC, Talley NJ. Mucosal inflammation as a potential etiological factor in irritable bowel syndrome: a systematic review. Journal of Gastroenterology. 2011;46(4):421–431.
    1. Kim DH, Cho YJ, Kim JH, Kim YB, Lee KJ. Stress-induced alterations in mast cell numbers and proteinaseactivated receptor-2 expression of the colon: role of corticotrophin-releasing factor. Journal of Korean Medical Science. 2010;25(9):1330–1335.
    1. Kawabata A, Matsunami M, Sekiguchi F. Gastrointestinal roles for proteinase-activated receptors in health and disease. British Journal of Pharmacology. 2008;153(supplement 1):S230–S240.
    1. Kayssi A, Amadesi S, Bautista F, Bunnett NW, Vanner S. Mechanisms of protease-activated receptor 2-evoked hyperexcitability of nociceptive neurons innervating the mouse colon. The Journal of Physiology. 2007;580(part 3):977–991.
    1. Gecse K, Róka R, Ferrier L, et al. Increased faecal serine protease activity in diarrhoeic IBS patients: a colonic lumenal factor impairing colonic permeability and sensitivity. Gut. 2008;57(5):591–598.
    1. Aloe L, Bracci-Laudiero L, Bonini S, Manni L. The expanding role of nerve growth factor: from neurotrophic activity to immunologic diseases. Allergy. 1997;52(9):883–894.
    1. Winston JH, Xu GY, Sarna SK. Adrenergic stimulation mediates visceral hypersensitivity to colorectal distension following heterotypic chronic stress. Gastroenterology. 2010;138(1):294–304.e3.
    1. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 2001;25(4):402–408.
    1. Xiang Z, Nilsson G. IgE receptor-mediated release of nerve growth factor by mast cells. Clinical and Experimental Allergy. 2000;30(10):1379–1386.
    1. Horigome K, Pryor JC, Bullock ED, Johnson EM. Mediator release from mast cells by nerve growth factor. Neurotrophin specificity and receptor mediation. The Journal of Biological Chemistry. 1993;268(20):14881–14887.
    1. Torrents D, Torres R, De Mora F, Vergara P. Antinerve growth factor treatment prevents intestinal dysmotility in Trichinella spiralis-infected rats. Journal of Pharmacology and Experimental Therapeutics. 2002;302(2):659–665.
    1. Galan A, Cervero F, Laird JM. Extracellular signaling-regulated kinase-1 and -2 (ERK 1/2) mediate referred hyperalgesia in a murine model of visceral pain. Molecular Brain Research. 2003;116(1-2):126–134.
    1. Cruz CD, Cruz F. The ERK 1 and 2 pathway in the nervous system: from basic aspects to possible clinical applications in pain and visceral dysfunction. Current Neuropharmacology. 2007;5(4):244–252.
    1. Lai HH, Qiu CS, Crock LW, et al. Activation of spinal extracellular signal-regulated kinases (ERK) 1/2 is associated with the development of visceral hyperalgesia of the bladder. Pain. 2011;152(9):2117–2124.
    1. Hong S, Fan J, Kemmerer ES, Evans S, Li Y, Wiley JW. Reciprocal changes in vanilloid (TRPV1) and endocannabinoid (CB1) receptors contribute to visceral hyperalgesia in the water avoidance stressed rat. Gut. 2009;58(2):202–210.
    1. Amadesi S, Nie J, Vergnolle N, et al. Protease-activated receptor 2 sensitizes the capsaicin receptor transient receptor potential vanilloid receptor 1 to induce hyperalgesia. The Journal of Neuroscience. 2004;24(18):4300–4312.

Source: PubMed

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