Application of in vivo imaging, a central technique for pre-clinical studies, in a model of pulmonary fibrosis

• Grant number: 14/09292-7
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree
• Start date: August 11, 2014
• End date: September 10, 2014
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Rodolfo de Paula Vieira
• Grantee: Ricardo Wesley Alberca Custódio
• Supervisor: Oliver Eickelberg
• Host Institution: Universidade Nove de Julho (UNINOVE). Campus Vergueiro. São Paulo , SP, Brazil
• Institution abroad: Ludwig Maximilian University of Munich (LMU Munich), Germany
• Associated to the scholarship: 12/21519-1 - Is the leukotrienes via contributes to the anti-inflammatory effects of aerobic exercise in asthma?, BP.MS

Abstract

In the last years a significant technological advance was reached in the direction of development of non-invasive methods in the animals' experimentation aiming to evaluate the inflammatory and the fibrotic process withouth the needing to submit the animals to euthanasia. In this way, new technologies, like the in vivo image systems are rising up, enabling a very detailed, even at molecular level, of the inflammatory and fibrotic process. These systems permit the capture and the analysis of fluorescence, multiplex fluorescence, luminescence, radioisotopic and also X-ray images. In the respiratory field these systems are being employed more and more, for instance, for detection of pulmonary levels of adenosine triphosphate (ATP) and for detection of the expression of NF-kB. In summary, these systems use basically marked/transfected cells with luciferin or fluorochromes for specific proteins and also transgenic mice for specific proteins, for instance, the GFP mice (green fluorescent protein). Thus, the present study aims to evaluate the pulmonary levels of ATP in a model of pulmonary fibrosis induced through the administration of bleomycin using an in vivo imaging technique. Male C57Bl/6 mice with 8 weeks (n = 8/group) will receive the bleomycin sulfate (1,5 U/kg) into the trachea and will be evaluated on days 0, 1, 2, 7, 14, 30 and 42 after the bleomycin administration. In addition to the in vivo detection of ATP levels will be also evaluated the number of inflammatory cells in the bronchoalveolar lavage (total and differential cells count), beyond the number of neutrophils and the quantification of collagen fibers in the lung parenchyma. For the in vivo imaging measurements of ATP, mice will receive the HEK293-pmeLUC cells into the trachea, which will allow us the bioluminescence through the luciferin. In a complementary way, will be also evaluated the ATP levels in the bronchoalveolar lavage by chemiluminescence, aiming to compare the methods for ATP measurements into the lungs, since that ATP is an important mediator of inflammatory and of fibrotic process in the lungs. (AU)

Scientific publications

(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)

ALBERCA-CUSTODIO, RICARDO WESLEY; GREIFFO, FLAVIA REGINA; MACKENZIE, BREANNE; OLIVEIRA-JUNIOR, MANOEL CARNEIRO; ANDRADE-SOUSA, ADILSON SANTOS; GRAUDENZ, GUSTAVO SILVEIRA; GOMES SANTOS, ANGELA BATISTA; DAMACENO-RODRIGUES, NILSA REGINA; CASTRO-FARIA-NETO, HUGO CAIRE; ARANTES-COSTA, FERNANDA MAGALHAES; et al. Aerobic Exercise Reduces Asthma Phenotype by Modulation of the Leukotriene Pathway. FRONTIERS IN IMMUNOLOGY, v. 7, JUN 14 2016. (12/15165-2, 14/23196-0, 13/23763-0, 12/23305-9, 14/14604-8, 12/21519-1, 14/09292-7)