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Highly selective CB2 receptor agonist A836339 has gastroprotective effect on experimentally induced gastric ulcers in mice

Abstract

Cannabinoid type 2 (CB2) receptors are distributed in central and peripheral tissues, including immunocytes and the gastrointestinal (GI) tract, suggesting that CB2 receptor agonists represent potential therapeutics in GI inflammatory states. In this study, we investigated the effect of highly selective CB2 agonist, A836339, on the development of gastric lesions. We used two models of gastric ulcer (GU) induced by ethanol (EtOH) and diclofenac. To confirm the involvement of CB2 receptors, a selective CB2 antagonist, AM630 was used. Clinical parameters for gastroprotection were assessed based on inhibition of the gastric lesion area. To investigate the anti-inflammatory effect of A836339, the expression of TNF-α and IL-1β was assessed. To establish the mechanism of gastroprotective action, catalase (CAT), superoxide dismutase (SOD) activity and H2O2 and glutathione (GSH) levels were measured. Moreover, expression of CB2 and cyclooxygenase-2 (COX-2) was characterized using immunohistochemistry (IHC). A836339 reduced ulcer index in a dose-dependent manner in both EtOH- and diclofenac-induced GU models. This effect was reversed by the CB2 antagonist AM630. Administration of A836339 reduced TNF-α and IL-1β levels in gastric tissue. Furthermore, A836339 exhibited potent anti-oxidant activity, as demonstrated by reduced H2O2 levels and increased CAT and SOD activities. IHC studies revealed a co-localization of CB2 receptors and COX-2 in the gastric tissue. Activation of CB2 receptors exhibited gastroprotective effect through enhancement of anti-oxidative pathways in the stomach. Activation of CB2 receptors may thus become a novel therapeutic approach in the treatment of GU.

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References

  • Alexander SPH, Benson HE, Faccenda E, Pawson AJ, Sharman JL, McGrath JC et al (2013) The concise guide to PHARMACOLOGY 2013/14: overview. Br J Pharmacol 170:1449–1458

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Alican İ, Coşkun T, Çorak A, Yeğen BÇ, Oktay Ş, Kurtel H (1995) Role of neutrophils in indomethacin-induced gastric mucosal lesions in rats. Inflamm Res 44:164–168

    CAS  Article  PubMed  Google Scholar 

  • Andrade SF, Antoniolli D, Comunello E, Cardoso LGV, Carvalho JCT, Bastos JK (2006) Antiulcerogenic activity of crude extract, fractions and populnoic acid isolated from Austroplenckia populnea (Celastraceae). Z Naturforsch C 61:329–333

    CAS  Article  PubMed  Google Scholar 

  • Antonisamy P, Arasu MV, Dhanasekaran M, Choi KC, Aravinthan A, Kim NS et al (2016) Protective effects of trigonelline against indomethacin-induced gastric ulcer in rats and potential underlying mechanisms. Food Funct 7:398–408

    CAS  Article  PubMed  Google Scholar 

  • Asmari AA, Omani SA, Otaibi MA, Abdulaaly A-AA, Elfaki I, Yahya KA et al (2014) Gastroprotective effect of minocycline in experimentally induced gastric ulcers in rats. Int J Clin Exp Med 7:586–596

    PubMed  PubMed Central  Google Scholar 

  • Blandizzi C, Tuccori M, Colucci R, Fornai M, Antonioli L, Ghisu N et al (2009) Role of coxibs in the strategies for gastrointestinal protection in patients requiring chronic non-steroidal anti-inflammatory therapy. Pharmacol Res 59:90–100

    CAS  Article  PubMed  Google Scholar 

  • Brzozowski T, Konturek PC, Konturek SJ, Kwiecién S, Pajdo R, Brzozowska I et al (1998) Involvement of endogenous cholecystokinin and somatostatin in gastroprotection induced by intraduodenal fat. J Clin Gastroenterol 27(Suppl 1):S125–S137

    Article  PubMed  Google Scholar 

  • Curtis MJ, Bond RA, Spina D, Ahluwalia A, Alexander SPA, Giembycz MA et al (2015) Experimental design and analysis and their reporting: new guidance for publication in BJP. Br J Pharmacol 172:3461–3471

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Fichna J, Bawa M, Thakur GA, Tichkule R, Makriyannis A, McCafferty D-M et al (2014a) Cannabinoids alleviate experimentally induced intestinal inflammation by acting at central and peripheral receptors. PLoS One 9:e109115

    Article  PubMed  PubMed Central  Google Scholar 

  • Fichna J, Sałaga M, Stuart J, Saur D, Sobczak M, Zatorski H et al (2014b) Selective inhibition of FAAH produces antidiarrheal and antinociceptive effect mediated by endocannabinoids and cannabinoid-like fatty acid amides. Neurogastroenterol Motil 26:470–481

    CAS  Article  PubMed  Google Scholar 

  • Fridovich I (1986) Biological effects of the superoxide radical. Arch Biochem Biophys 247:1–11

    CAS  Article  PubMed  Google Scholar 

  • Germanò MP, D’Angelo V, Mondello MR, Pergolizzi S, Capasso F, Capasso R et al (2001) Cannabinoid CB1-mediated inhibition of stress-induced gastric ulcers in rats. Naunyn Schmiedeberg's Arch Pharmacol 363:241–244

    Article  Google Scholar 

  • Glavin GB, Szabo S (1992) Experimental gastric mucosal injury: laboratory models reveal mechanisms of pathogenesis and new therapeutic strategies. FASEB J 6:825–831

    CAS  PubMed  Google Scholar 

  • Graham ES, Ashton JC, Glass M (2009) Cannabinoid receptors: a brief history and “what’s hot”. Front Biosci Landmark Ed 14:944–957

    CAS  Article  PubMed  Google Scholar 

  • Gui H, Sun Y, Luo Z-M, Su D-F, Dai S-M, Liu X (2013) Cannabinoid receptor 2 protects against acute experimental sepsis in mice. Mediat Inflamm 2013:1–10

    Article  Google Scholar 

  • Howlett AC, Barth F, Bonner TI, Cabral G, Casellas P, Devane WA et al (2002) International Union of Pharmacology XXVII Classification of cannabinoid receptors. Pharmacol Rev 54:161–202

    CAS  Article  PubMed  Google Scholar 

  • Hsieh GC, Pai M, Chandran P, Hooker BA, Zhu CZ, Salyers AK et al (2011) Central and peripheral sites of action for CB2 receptor mediated analgesic activity in chronic inflammatory and neuropathic pain models in rats. Br J Pharmacol 162:428–440

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Ihenetu K, Molleman A, Parsons ME, Whelan CJ (2003) Inhibition of interleukin-8 release in the human colonic epithelial cell line HT-29 by cannabinoids. Eur J Pharmacol 458:207–215

    CAS  Article  PubMed  Google Scholar 

  • Izzo AA, Sharkey KA (2010) Cannabinoids and the gut: new developments and emerging concepts. Pharmacol Ther 126:21–38

    CAS  Article  PubMed  Google Scholar 

  • Kimball ES, Schneider CR, Wallace NH, Hornby PJ (2006) Agonists of cannabinoid receptor 1 and 2 inhibit experimental colitis induced by oil of mustard and by dextran sulfate sodium. AJP Gastrointest Liver Physiol 291:G364–G371

    CAS  Article  Google Scholar 

  • Kinsey SG, Nomura DK, O’Neal ST, Long JZ, Mahadevan A, Cravatt BF et al (2011) Inhibition of monoacylglycerol lipase attenuates nonsteroidal anti-inflammatory drug-induced gastric hemorrhages in mice. J Pharmacol Exp Ther 338:795–802

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Konturek PC, Celinski K, Slomka M, Cichoz-Lach H, Burnat G, Naegel A et al (2008) Melatonin and its precursor L-tryptophan prevent acute gastric mucosal damage induced by aspirin in humans. J Physiol Pharmacol 59(Suppl 2):67–75

    PubMed  Google Scholar 

  • Laine L, Weinstein WM (1988) Histology of alcoholic hemorrhagic “gastritis”: a prospective evaluation. Gastroenterology 94:1254–1262

    CAS  Article  PubMed  Google Scholar 

  • Laloo D, Prasad SK, Krishnamurthy S, Hemalatha S (2013) Gastroprotective activity of ethanolic root extract of Potentilla fulgens Wall. ex Hook. J Ethnopharmacol 146:505–514

    Article  PubMed  Google Scholar 

  • Lee M-Y, Shin I-S, Jeon W-Y, Seo C-S, Ha H, Huh J-I et al (2012) Protective effect of Bojungikki-tang, a traditional herbal formula, against alcohol-induced gastric injury in rats. J Ethnopharmacol 142:346–353

    Article  PubMed  Google Scholar 

  • McGrath J, Drummond G, McLachlan E, Kilkenny C, Wainwright C (2010) Guidelines for reporting experiments involving animals: the ARRIVE guidelines. Br J Pharmacol 160:1573–1576

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Munro S, Thomas KL, Abu-Shaar M (1993) Molecular characterization of a peripheral receptor for cannabinoids. Nature 365:61–65

    CAS  Article  PubMed  Google Scholar 

  • Naidu PS, Booker L, Cravatt BF, Lichtman AH (2009) Synergy between enzyme inhibitors of fatty acid amide hydrolase and cyclooxygenase in visceral nociception. J Pharmacol Exp Ther 329:48–56

    CAS  Article  PubMed  Google Scholar 

  • Pertwee RG (2006) The pharmacology of cannabinoid receptors and their ligands: an overview. Int J Obes 30:S13–S18

    CAS  Article  Google Scholar 

  • Piechota-Polanczyk A, Fichna J (2014) Review article: the role of oxidative stress in pathogenesis and treatment of inflammatory bowel diseases. Naunyn Schmiedeberg's Arch Pharmacol 387:605–620

    CAS  Article  Google Scholar 

  • Robert A, Nezamis JE, Lancaster C, Hanchar AJ (1979) Cytoprotection by prostaglandins in rats. Prevention of gastric necrosis produced by alcohol, HCl, NaOH, hypertonic NaCl, and thermal injury. Gastroenterology 77:433–443

    CAS  PubMed  Google Scholar 

  • Rutkowska M, Fereniec-Gołebiewska L, Rutkowska M (2006) ACEA (arachidonyl-2-chloroethylamide), the selective cannabinoid CB 1 receptor agonist, protects against aspirin-induced gastric ulceration. Pharmazie 61:341–342

    CAS  PubMed  Google Scholar 

  • Rutkowska M, Fereniec-Gołebiewska L (2009) Involvement of nitric oxide in the gastroprotective effect of ACEA, a selective cannabinoid CB1 receptor agonist, on aspirin-induced gastric ulceration. Pharmazie 64:595–597

    CAS  PubMed  Google Scholar 

  • Sałaga M, Lewandowska U, Sosnowska D, Zakrzewski PK, Cygankiewicz AI, Piechota-Polańczyk A et al (2014a) Polyphenol extract from evening primrose pomace alleviates experimental colitis after intracolonic and oral administration in mice. Naunyn Schmiedeberg's Arch Pharmacol 387:1069–1078

    Article  Google Scholar 

  • Sałaga M, Mokrowiecka A, Zakrzewski PK, Cygankiewicz A, Leishman E, Sobczak M et al (2014b) Experimental colitis in mice is attenuated by changes in the levels of endocannabinoid metabolites induced by selective inhibition of fatty acid amide hydrolase (FAAH). J. Crohn’s Colitis 8:998–1009

    Article  Google Scholar 

  • Sałaga M, Sobczak M, Fichna J (2014c) Inhibition of fatty acid amide hydrolase (FAAH) as a novel therapeutic strategy in the treatment of pain and inflammatory diseases in the gastrointestinal tract. Eur J Pharm Sci 52:173–179

    Article  PubMed  Google Scholar 

  • Sedlak J, Lindsay RH (1968) Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Anal Biochem 25:192–205

    CAS  Article  PubMed  Google Scholar 

  • Shimoyama AT, Santin JR, Machado ID, de Oliveira e Silva AM, de Melo ILP, Mancini-Filho J et al (2013) Antiulcerogenic activity of chlorogenic acid in different models of gastric ulcer. Naunyn Schmiedeberg's Arch Pharmacol 386:5–14

    CAS  Article  Google Scholar 

  • Shujaa N, Zadori ZS, Ronai AZ, Barna I, Mergl Z, Mozes MM et al (2009) Analysis of the effect of neuropeptides and cannabinoids in gastric mucosal defense initiated centrally in the rat. J Physiol Pharmacol 60(Suppl 7):93–100

    PubMed  Google Scholar 

  • Singh LP, Mishra A, Saha D, Swarnakar S (2011) Doxycycline blocks gastric ulcer by regulating matrix metalloproteinase-2 activity and oxidative stress. World J Gastroenterol 17:3310–3321

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Storr M, Gaffal E, Saur D, Schusdziarra V, Allescher HD (2002) Effect of cannabinoids on neural transmission in rat gastric fundus. Can J Physiol Pharmacol 80:67–76

    CAS  Article  PubMed  Google Scholar 

  • Tanaka A, Araki H, Hase S, Komoike Y, Takeuchi K (2002a) Up-regulation of COX-2 by inhibition of COX-1 in the rat: a key to NSAID-induced gastric injury. Aliment Pharmacol Ther 16(Suppl 2):90–101

    CAS  Article  PubMed  Google Scholar 

  • Tanaka A, Hase S, Miyazawa T, Ohno R, Takeuchi K (2002b) Role of cyclooxygenase (COX)-1 and COX-2 inhibition in nonsteroidal anti-inflammatory drug-induced intestinal damage in rats: relation to various pathogenic events. J Pharmacol Exp Ther 303:1248–1254

    CAS  Article  PubMed  Google Scholar 

  • Tanaka A, Hase S, Miyazawa T, Takeuchi K (2002c) Up-regulation of cyclooxygenase-2 by inhibition of cyclooxygenase-1: a key to nonsteroidal anti-inflammatory drug-induced intestinal damage. J Pharmacol Exp Ther 300:754–761

    CAS  Article  PubMed  Google Scholar 

  • Tuluce Y, Ozkol H, Koyuncu I, Ine H (2011) Gastroprotective effect of small centaury (Centaurium erythraea L) on aspirin-induced gastric damage in rats. Toxicol Ind Health 27:760–768

    Article  PubMed  Google Scholar 

  • Wallace JL, McKnight W, Reuter BK, Vergnolle N (2000) NSAID-induced gastric damage in rats: requirement for inhibition of both cyclooxygenase 1 and 2. Gastroenterology 119:706–714

    CAS  Article  PubMed  Google Scholar 

  • Wang T, Zhao S, Wang Y, Yang Y, Yao L, Chu L et al (2014) Protective effects of escin against indomethacin-induced gastric ulcer in mice. Toxicol Mech Methods 24:560–566

    Article  PubMed  Google Scholar 

  • Warzecha Z, Dembinski A, Ceranowicz P, Dembinski M, Cieszkowski J, Kownacki P et al (2011) Role of sensory nerves in gastroprotective effect of anandamide in rats. J Physiol Pharmacol 62:207–217

    CAS  PubMed  Google Scholar 

  • Wright K, Rooney N, Feeney M, Tate J, Robertson D, Welham M et al (2005) Differential expression of cannabinoid receptors in the human colon: cannabinoids promote epithelial wound healing. Gastroenterology 129:437–453

    Article  PubMed  Google Scholar 

  • Yao BB, Hsieh G, Daza AV, Fan Y, Grayson GK, Garrison TR et al (2009) Characterization of a cannabinoid CB2 receptor-selective agonist, A-836339 [2,2,3,3-tetramethyl-cyclopropanecarboxylic acid [3-(2-methoxy-ethyl)-4,5-dimethyl-3H-thiazol-(2Z)-ylidene]-amide], using in vitro pharmacological assays, in vivo pain models, and pharmacological magnetic resonance imaging. J Pharmacol Exp Ther 328:141–151

    CAS  Article  PubMed  Google Scholar 

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Acknowledgments

This study was supported by the Iuventus Plus program of the Polish Ministry of Science and Higher Education (IP2015 068774 to MS), with grants from the Medical University of Lodz (502-03/1-156-04/502-14-298 to M Salaga, 502-03/1-156-02/502-14-141 to MZ and 503/1-156-04/503-11-001 to JF) and the National Science Centre (UMO-2013/11/N/NZ7/02354 to M Salaga, UMO-2013/11/N/NZ7/00724 to MZ, UMO-2013/11/B/NZ7/01301 to JF and UMO-2014/13/B/NZ4/01179 to JF). Hubert Zatorski is the recipient of Diamentowy Grant Program of the Polish Ministry of Science and Higher Education (No. 0202/DIA/2015/44).

Authors’ contributions

M Salaga, H Zatorski, M Zielinska, P Mosinska, J-P Timmermans and R Kordek performed the research. M Salaga, H Zatorski and J Fichna designed the research study. M Storr and J Fichna contributed essential tools and reagents. M Salaga, H Zatorski, M Zielińska, J-P Timmermans and R Kordek analysed the data. J-P Timmermans and M Storr critically reviewed the manuscript. M Salaga, H Zatorski and J Fichna wrote the paper.

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Correspondence to J. Fichna.

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The study was carried out in accordance with the recommendations described in the Guide for the Care and Use of Laboratory Animals of the Medical University of Lodz, Poland. All experiments on animals were approved by the Local Ethical Committee for Animal Experiments (Protocol #17/LB654/2013).

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The authors declare that they have no conflict of interest.

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Salaga, M., Zatorski, H., Zielińska, M. et al. Highly selective CB2 receptor agonist A836339 has gastroprotective effect on experimentally induced gastric ulcers in mice. Naunyn-Schmiedeberg's Arch Pharmacol 390, 1015–1027 (2017). https://doi.org/10.1007/s00210-017-1402-3

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  • DOI: https://doi.org/10.1007/s00210-017-1402-3

Keywords

  • Gastric ulcer
  • Endocannabinoid system
  • CB2 receptors
  • Oxidative stress