Study of the effect of experimental ulcerative colitis on the enteric nervous system of P2X7 receptor deficient mice (P2X7-/-)

• Grant number: 18/07862-1
• Support Opportunities: Regular Research Grants
• Duration: March 01, 2019 - August 31, 2021
• Field of knowledge: Biological Sciences - Morphology - Anatomy
• Principal Investigator: Patricia Castelucci
• Grantee: Patricia Castelucci
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

5'-adenosine triphosphate (ATP) is known as a neurotransmitter and its receptors are the P2X1-7. The presence of P2X2, 3, 7 purinergic receptor in the enteric nervous system has been demonstrated. Ulcerative colitis and Crohn's disease present pathophysiological processes, necrosis of the gut and the enteric neurons and enteric glial cells. It has been demonstrated the participation of the P2X7 receptor in the inflammatory process and cell necrosis. This project aims to study the role of the P2X7 receptor in the myenteric plexus and enteric glial cells in the ulcerative colitis of Wild Type and mice deficient in the P2X7 receptor (P2X7 -/-, P2X7KO). 2, 4, 6, trinitrobenzene sulfonic acid (TNBS) will be injected into the large intestine of C57BL/6 mice and P2X7KO, in the sham group will be injected vehicle. It will be analyzed: a) colocalization of the P2X7 receptor with nitric oxide synthase (NOS), choline acetyl transferase (ChAT), PGP9.5 (pan neuronal) and S100B/GFAP (mark for enteric glial cells); b) neuron per ganglia and area of the cellular profile (micrometro2) of the immunoreactive neurons to the P2X7 receptor, NOS, ChAT, PGP9.5 and enteric glias; c) histology of the distal colon; d) intestinal motility; e) the presence of inflammatory cytokines; f) the MAPK signaling pathways: ERK-1/2, p38 MAPK and caspase-3; g) protein and gene expression of the P2X7 receptor.For this purpose, double- and triple-immunofluorescence techniques will be used, qualitative and quantitative analyzes will be obtained from the fluorescence microscopes and Confocal Laser Scanning. For the study of protein expression, will use Western Blotting; for the investigation of intestinal motility will be used organ bath system; for the study of the cytokines will be used the methods of Elisa and for the morphological analysis of the large intestine will be used the method of conventional histology. After that, statistical analysis will be done. The project has the justification to elucidate the mechanisms involved in neuronal death by the P2X7 receptor in the intestinal inflammation model and to verify the potential therapeutic targets. (AU)

Scientific publications

(8)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MENDES, CRISTINA EUSEBIO; PALOMBIT, KELLY; PEREIRA, THAIRA THALITA ALVES; MAGALHAES, HENRIQUE INHAUSER RICETI; CAETANO, MARCOS ANTONTIO FERREIRA; CASTELUCCI, PATRICIA. Effects of probenecid and brilliant blue G on rat enteric glial cells following intestinal ischemia and reperfusion. ACTA HISTOCHEMICA, v. 125, n. 1, p. 13-pg., 2022-12-07. (15/22299-3, 12/00259-1, 18/07862-1, 14/25927-2)

SOUZA, ROBERTA FIGUEIROA; CAETANO, MARCOS ANTONIO FERREIRA; MAGALHAES, HENRIQUE INHAUSER RICETI; CASTELUCCI, PATRICIA. Study of tumor necrosis factor receptor in the inflammatory bowel disease. WORLD JOURNAL OF GASTROENTEROLOGY, v. 29, n. 18, p. 14-pg., 2023-05-14. (18/07862-1, 14/25927-2)

EVANGELINELLIS, MARIA MUNHOZ; SOUZA, ROBERTA FIGUEIROA; MENDES, CRISTINA EUSEBIO; CASTELUCCI, PATRICIA. Effects of a P2X7 receptor antagonist on myenteric neurons in the distal colon of an experimental rat model of ulcerative colitis. Histochemistry and Cell Biology, v. 157, n. 1, OCT 2021. (18/07862-1, 12/00259-1, 14/25927-2)

SANTOS, ANA A. Q. A.; COSTA, DEIZIANE V. S.; FOSCHETTI, DANIELLE A.; DUARTE, ANTONIELLA S. G.; MARTINS, CONCEICAO S.; SOARES, PEDRO M. G.; CASTELUCCI, PATRICIA; BRITO, GERLY A. C.. P2X7 receptor blockade decreases inflammation, apoptosis, and enteric neuron loss during Clostridioides difficile toxin A-induced ileitis in mice. WORLD JOURNAL OF GASTROENTEROLOGY, v. 28, n. 30, p. 14-pg., 2022-08-14. (14/25927-2, 18/07862-1)

CAETANO, MARCOS A. F.; MAGALHAES, HENRIQUE I. R.; DUARTE, JHENIFFER R. L.; CONCEICAO, LAURA B.; CASTELUCCI, PATRICIA. Butyrate Protects Myenteric Neurons Loss in Mice Following Experimental Ulcerative Colitis. CELLS, v. 12, n. 13, p. 23-pg., 2023-07-01. (18/07862-1, 14/25927-2)

MAGALHAES, HENRIQUE INHAUSER RICETI; MACHADO, FELIPE ALEXANDRE; SOUZA, ROBERTA FIGUEIROA; CAETANO, MARCOS ANTONIO FERREIRA; FIGLIUOLO, VANESSA RIBEIRO; COUTINHO-SILVA, ROBSON; CASTELUCCI, PATRICIA. Study of the roles of caspase-3 and nuclear factor kappa B in myenteric neurons in a P2X7 receptor knockout mouse model of ulcerative colitis. WORLD JOURNAL OF GASTROENTEROLOGY, v. 29, n. 22, p. 29-pg., 2023-06-14. (18/07862-1, 14/25927-2)

MACHADO, FELIPE ALEXANDRE; SOUZA, ROBERTA FIGUEIROA; FIGLIUOLO, VANESSA RIBEIRO; COUTINHO-SILVA, ROBSON; CASTELUCCI, PATRICIA. Effects of experimental ulcerative colitis on myenteric neurons in P2X7-knockout mice. Histochemistry and Cell Biology, v. 160, n. 4, p. 19-pg., 2023-06-12. (18/07862-1, 14/25927-2)

CAETANO, MARCOS ANTONIO FERREIRA; CASTELUCCI, PATRICIA. Role of short chain fatty acids in gut health and possible therapeutic approaches in inflammatory bowel diseases. WORLD JOURNAL OF CLINICAL CASES, v. 10, n. 28, p. 19-pg., 2022-10-06. (14/25927-2, 18/07862-1)