{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T03:16:21Z","timestamp":1790046981417,"version":"4.0.1"},"reference-count":48,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2021,10,28]],"date-time":"2021-10-28T00:00:00Z","timestamp":1635379200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"National Science Center","doi-asserted-by":"publisher","award":["2018\/29\/N\/NZ4\/00348"],"award-info":[{"award-number":["2018\/29\/N\/NZ4\/00348"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Side effects associated with nonsteroidal anti-inflammatory drugs (NSAIDs) treatment are a serious limitation of their use in anti-inflammatory therapy. The negative effects of taking NSAIDs include abdominal pain, indigestion nausea as well as serious complications such as bleeding and perforation. The enteric nervous system is involved in regulation of gastrointestinal functions through the release of neurotransmitters. The present study was designed to determine, for the first time, the changes in pituitary adenylate cyclase-activating polypeptide (PACAP), substance P (SP) and galanin (GAL) expression in porcine jejunum after long-term treatment with aspirin, indomethacin and naproxen. The study was performed on 16 immature pigs. The animals were randomly divided into four experimental groups: control, aspirin, indomethacin and naproxen. Control animals were given empty gelatin capsules, while animals in the test groups received selected NSAIDs for 28 days. Next, animals from each group were euthanized. Frozen sections were prepared from collected jejunum and subjected to double immunofluorescence staining. NSAIDs supplementation caused a significant increase in the population of PACAP-, SP- and GAL-containing enteric neurons in the porcine jejunum. Our results suggest the participation of the selected neurotransmitters in regulatory processes of the gastrointestinal function and may indicate the direct toxic effect of NSAIDs on the ENS neurons.<\/jats:p>","DOI":"10.3390\/ijms222111689","type":"journal-article","created":{"date-parts":[[2021,10,28]],"date-time":"2021-10-28T23:51:36Z","timestamp":1635465096000},"page":"11689","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Effect of NSAIDs Supplementation on the PACAP-, SP- and GAL-Immunoreactive Neurons in the Porcine Jejunum"],"prefix":"10.3390","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7233-4187","authenticated-orcid":false,"given":"Marta","family":"Brzozowska","sequence":"first","affiliation":[{"name":"Department of Clinical Physiology, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Oczapowskiego Str. 13, 10-718 Olsztyn, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Jana","sequence":"additional","affiliation":[{"name":"Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Tuwima Str. 10, 10-748 Olsztyn, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaros\u0142aw","family":"Ca\u0142ka","sequence":"additional","affiliation":[{"name":"Department of Clinical Physiology, Faculty of Veterinary Medicine, University of Warmia and Mazury in Olsztyn, Oczapowskiego Str. 13, 10-718 Olsztyn, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,10,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1111\/j.1743-3150.2004.00475.x","article-title":"Lessons from the porcine enteric nervous system","volume":"16","author":"Brown","year":"2004","journal-title":"Neurogastroenterol. Motil."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"286","DOI":"10.1038\/nrgastro.2012.32","article-title":"The enteric nervous system and neurogastroenterology","volume":"9","author":"Furness","year":"2012","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"567","DOI":"10.1016\/0006-291X(89)91757-9","article-title":"Isolation of a novel 38 residue-hypothalamic polypeptide which stimulates adenylate cyclase in pituitary cells","volume":"164","author":"Miyata","year":"1989","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1016\/0006-291X(90)92140-U","article-title":"Isolation of a neuropeptide corresponding to the N-terminal 27 residues of the pituitary adenylate cyclase activating polypeptide with 38 residues (PACAP38)","volume":"170","author":"Miyata","year":"1990","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1124\/pr.109.001370","article-title":"Pituitary adenylate cyclase-activating polypeptide and its receptors: 20 years after the discovery","volume":"61","author":"Vaudry","year":"2009","journal-title":"Pharmacol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"377","DOI":"10.3389\/fendo.2020.00377","article-title":"Protective Effects of PACAP in Peripheral Organs","volume":"11","author":"Toth","year":"2020","journal-title":"Front. Endocrinol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1007\/s00018-007-7148-1","article-title":"Tachykinins and their functions in the gastrointestinal tract","volume":"65","author":"Shimizu","year":"2008","journal-title":"Cell Mol. Life Sci."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Bombardi, C., Rambaldi, A.M., Galiazzo, G., Giancola, F., Gra\u00efc, J.M., Salamanca, G., Cozzi, B., and Chiocchetti, R. (2021). Nitrergic and Substance P Immunoreactive Neurons in the Enteric Nervous System of the Bottlenose Dolphin (Tursiops truncatus) Intestine. Animals, 11.","DOI":"10.3390\/ani11041057"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"602070","DOI":"10.3389\/fimmu.2020.602070","article-title":"Review: Occurrence and Distribution of Galanin in the Physiological and Inflammatory States in the Mammalian Gastrointestinal Tract","volume":"11","author":"Brzozowska","year":"2021","journal-title":"Front. Immunol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/j.pharmthera.2007.05.009","article-title":"The galanin peptide family: Receptor pharmacology, pleiotropic biological actions, and implications in health and disease","volume":"115","author":"Lang","year":"2007","journal-title":"Pharmacol. Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.autneu.2007.04.005","article-title":"Innervation of the gastrointestinal tract: Patterns of aging","volume":"136","author":"Phillips","year":"2007","journal-title":"Auton. Neurosci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1038\/s41366-018-0166-2","article-title":"Interplay between colonic inflammation and tachykininergic pathways in the onset of colonic dysmotility in a mouse model of diet-induced obesity","volume":"43","author":"Antonioli","year":"2019","journal-title":"Int. J. Obes."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1556","DOI":"10.1007\/s12031-019-01473-y","article-title":"Changes in the Neurochemical Characterization of Enteric Neurons in the Porcine Duodenum After Administration of Low-Dose Salmonella Enteritidis Lipopolysaccharides","volume":"71","author":"Rytel","year":"2021","journal-title":"J. Mol. Neurosci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1136\/gut.40.6.767","article-title":"Neurochemical coding in the small intestine of patients with Crohn\u2019s disease","volume":"40","author":"Belai","year":"1997","journal-title":"Gut"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1017\/S0031182000066415","article-title":"Nippostrongylus brasiliensis infection evokes neuronal abnormalities and alterations in neurally regulated electrolyte transport in rat jejunum","volume":"113","author":"Masson","year":"1996","journal-title":"Parasitology"},{"key":"ref_16","first-page":"475","article-title":"Somatostatin, substance P and calcitonin gene-related peptide-positive intramural nerve structures of the human large intestine affected by carcinoma","volume":"30","author":"Godlewski","year":"2010","journal-title":"Folia Histochem. Cytobiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1096","DOI":"10.1136\/gut.2003.021154","article-title":"Bacillary dysentery as a causative factor of irritable bowel syndrome and its pathogenesis","volume":"53","author":"Wang","year":"2004","journal-title":"Gut."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e13580","DOI":"10.1111\/nmo.13580","article-title":"Neurochemical characterization of the enteric neurons within the porcine jejunum in physiological conditions and under the influence of bisphenol A (BPA)","volume":"31","author":"Szymanska","year":"2019","journal-title":"Neurogastroenterol. Motil."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"836","DOI":"10.1136\/gut.42.6.836","article-title":"Enteric neuronal plasticity and a reduced number of interstitial cells of Cajal in hypertrophic rat ileum","volume":"42","author":"Ekblad","year":"1998","journal-title":"Gut"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"114147","DOI":"10.1016\/j.bcp.2020.114147","article-title":"Non-steroidal anti-inflammatory drugs (NSAIDs) and organ damage: A current perspective","volume":"180","author":"Bindu","year":"2020","journal-title":"Biochem. Pharmacol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"216","DOI":"10.14740\/gr658w","article-title":"Cross-Sectional Study for Prevalence of Non-Steroidal Anti-Inflammatory Drug-Induced Gastrointestinal Cardiac and Renal Complications in India: Interim Report","volume":"8","author":"Chatterjee","year":"2015","journal-title":"Gastroenterol. Res."},{"key":"ref_22","first-page":"218","article-title":"Frequency of NSAID induced peptic ulcer disease","volume":"56","author":"Hamid","year":"2006","journal-title":"J. Pak. Med. Assoc."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"151425","DOI":"10.1016\/j.aanat.2019.151425","article-title":"Long-term treatment with naproxen changes the chemical coding of the porcine intramural duodenum neurons","volume":"227","author":"Czajkowska","year":"2020","journal-title":"Ann. Anat."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1007\/s00535-009-0102-2","article-title":"Present status and strategy of NSAIDs-induced small bowel injury","volume":"44","author":"Higuchi","year":"2009","journal-title":"J. Gastroenterol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"564457","DOI":"10.3389\/fphar.2020.564457","article-title":"Neurochemistry of Enteric Neurons Following Prolonged Indomethacin Administration in the Porcine Duodenum","volume":"11","author":"Czajkowska","year":"2020","journal-title":"Front. Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"90","DOI":"10.3389\/fendo.2018.00090","article-title":"Presence and Effects of Pituitary Adenylate Cyclase Activating Polypeptide Under Physiological and Pathological Conditions in the Stomach","volume":"9","author":"Reglodi","year":"2018","journal-title":"Front. Endocrinol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1016\/j.pneurobio.2015.10.004","article-title":"Novel tactics for neuroprotection in Parkinson\u2019s disease: Role of antibiotics, polyphenols and neuropeptides","volume":"155","author":"Reglodi","year":"2017","journal-title":"Prog. Neurobiol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1007\/s12031-014-0273-z","article-title":"PACAP27 is protective against tat-induced neurotoxicity","volume":"54","author":"Rozzi","year":"2014","journal-title":"J. Mol. Neurosci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/S0024-3205(98)00280-X","article-title":"Effects of pituitary adenylate cyclase activating polypeptide-27 on alkaline secretory and mucosal ulcerogenic responses in rat duodenum","volume":"63","author":"Yagi","year":"1998","journal-title":"Life Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2012","DOI":"10.1073\/pnas.0812257106","article-title":"Pituitary adenylyl cyclase-activating polypeptide is an intrinsic regulator of Treg abundance and protects against experimental autoimmune encephalomyelitis","volume":"106","author":"Tan","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1097\/MED.0b013e328342568a","article-title":"VIP and PACAP: Recent insights into their functions\/roles in physiology and disease from molecular and genetic studies","volume":"18","author":"Moody","year":"2011","journal-title":"Curr. Opin. Endocrinol. Diabetes Obes."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1111\/j.1749-6632.2010.05512.x","article-title":"Pituitary adenylate cyclase activating polypeptide in the retina: Focus on the retinoprotective effects","volume":"1200","author":"Atlasz","year":"2010","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1803","DOI":"10.1002\/ijc.23308","article-title":"Induction of colitis and rapid development of colorectal tumors in mice deficient in the neuropeptide PACAP","volume":"122","author":"Nemetz","year":"2008","journal-title":"Int. J. Cancer"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/S0167-0115(02)00194-5","article-title":"PAC1 and PACAP expression, signaling, and effect on the growth of HCT8, human colonic tumor cells","volume":"109","author":"Le","year":"2002","journal-title":"Regul. Pept."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/S0898-6568(97)00067-3","article-title":"Differential expression and function of PACAP and VIP receptors in four human colonic adenocarcinoma cell lines","volume":"10","author":"Meunier","year":"1998","journal-title":"Cell Signal."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1002\/eji.201848061","article-title":"The role of galanin in the differentiation of mucosal mast cells in mice","volume":"50","author":"Yamaguchi","year":"2020","journal-title":"Eur J. Immunol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1016\/j.regpep.2006.12.029","article-title":"Chronic administration of galanin attenuates the TNBS-induced colitis in rats","volume":"141","author":"Talero","year":"2007","journal-title":"Regul. Pept."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1048","DOI":"10.1038\/79563","article-title":"Galanin-1 receptor up-regulation mediates the excess colonic fluid production caused by infection with enteric pathogens","volume":"6","author":"Matkowskyj","year":"2000","journal-title":"Nat. Med."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1404","DOI":"10.1016\/j.intimp.2006.04.016","article-title":"Galanin in the trinitrobenzene sulfonic acid rat model of experimental colitis","volume":"6","author":"Talero","year":"2006","journal-title":"Int. Immunopharmacol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"179","DOI":"10.5603\/FHC.a2019.0020","article-title":"Increased expression of CART, nNOS, VIP, PACAP, SP and GAL in enteric neurons of the porcine stomach prepyloric region following hydrochloric acid infusion","volume":"57","author":"Calka","year":"2019","journal-title":"Folia Histochem. Cytobiol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Palus, K., Makowska, K., and Ca\u0142ka, J. (2019). Alterations in Galanin-Like Immunoreactivity in the Enteric Nervous System of the Porcine Stomach Following Acrylamide Supplementation. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20133345"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1159\/000007504","article-title":"Implications of tachykinins and calcitonin gene-related peptide in inflammatory bowel disease","volume":"59","author":"Holzer","year":"1998","journal-title":"Digestion"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1016\/S1357-2725(01)00031-0","article-title":"Substance P","volume":"33","author":"Harrison","year":"2001","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1556\/004.2020.00028","article-title":"Changes in the expression of substance P in nerve fibres of the colonic mucosa in dogs suffering from inflammatory bowel disease","volume":"68","author":"Rychlik","year":"2020","journal-title":"Acta Vet. Hung."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1159\/000084625","article-title":"Gastric perception thresholds are low and sensory neuropeptide levels high in helicobacter pylori-positive functional dyspepsia","volume":"71","author":"Wollenberg","year":"2005","journal-title":"Digestion"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"2864","DOI":"10.1007\/s10620-014-3263-7","article-title":"Tachykinin peptide, substance P, and its receptor NK-1R play an important role in alimentary tract mucosal inflammation during cytotoxic therapy","volume":"59","author":"Satheeshkumar","year":"2014","journal-title":"Dig. Dis. Sci."},{"key":"ref_47","first-page":"75","article-title":"Substance P induces TNF-alpha and IL-6 production through NF kappa B in peritoneal mast cells. Biochim Biophys","volume":"1643","author":"Azzolina","year":"2003","journal-title":"Acta Mol. Cell Res."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Rz\u0105p, D., Czajkowska, M., and Ca\u0142ka, J. (2020). Neurochemical Plasticity of nNOS-, VIP- and CART-Immunoreactive Neurons Following Prolonged Acetylsalicylic Acid Supplementation in the Porcine Jejunum. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21062157"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/21\/11689\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:22:08Z","timestamp":1760167328000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/21\/11689"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,28]]},"references-count":48,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2021,11]]}},"alternative-id":["ijms222111689"],"URL":"https:\/\/doi.org\/10.3390\/ijms222111689","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,28]]}}}