DANIELA APARECIDA DE BRITO CERVILHA
Projetos de Pesquisa
Unidades Organizacionais
LIM/20 - Laboratório de Terapêutica Experimental, Hospital das Clínicas, Faculdade de Medicina
13 resultados
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Agora exibindo 1 - 10 de 13
- The tick-derived rBmTI-A protease inhibitor attenuates the histological and functional changes induced by cigarette smoke exposure(2018) LOURENCO, Juliana D.; ITO, Juliana T.; CERVILHA, Daniela A. B.; SALES, Davi S.; RIANI, Alyne; SUEHIRO, Camila L.; GENARO, Isabella S.; DURAN, Adriana; PUZER, Luciano; MARTINS, Milton A.; SASAKI, Sergio D.; LOPES, Fernanda D. T. Q. S.Introduction. Smoking is the main risk factor for chronic obstructive pulmonary disease development and cigarette smoke (CS) exposure is considered an important approach to reproduce in rodents this human disease. We have previously shown that in an elastase induced model of emphysema, the administration of a protease inhibitor (rBmTI-A) prevented and attenuated tissue destruction in mice. Thus, in this study we aimed to verify the effects of rBmTI-A administration on the physiopathological mechanisms of CS induced emphysema. Methods. Mice (C57BL/6) were exposed to CS or room air for 12 weeks. In this period, 3 nasal instillations of rBmTI-A inhibitor or its vehicle were performed. After euthanasia, respiratory mechanics were evaluated and lungs removed for analysis of mean linear intercept, volume proportion of collagen and elastic fibers, density of polymorphonuclear cells, macrophages, and density of positive cells for MMP-12, MMP-9, TIMP-1 and gp91phox. Results. The rBmTI-A administration improved tissue elastance, decreased alveolar enlargement and collagen fibers accumulation to control levels and attenuated elastic fibers accumulation in animals exposed to CS. There was an increase of MMP-12, MMP-9 and macrophages in CS groups and the rBmTIA only decreased the number of MMP-12 positive cells. Also, we demonstrated an increase in gp91phox in CS treated group and in TIMP-1 levels in both rBmTI-A treated groups. Conclusion. In summary, the rBmTI-A administration attenuated emphysema development by an increase of gp91phox and TIMP-1, accompanied by a decrease in MMP-12 levels.
conferenceObject Cigarette Smoke Exposure Combined To LPS Instillation: A New Experimental Model Of Exacerbation In Chronic Obstructive Pulmonary Disease In Mice(2016) CERVILHA, D. A. B.; ITO, J. T.; LOURENCO, J. D.; OLIVO, C. R.; SALES, D. S.; SARAIVA, B. M.; OLIVEIRA-JUNIOR, M. C.; MARTINS, M. A.; VIEIRA, R. P.; LOPES, F. D. T. Q. S.- Temporal analysis of the intracellular signaling pathways involved in Th17/Treg response in COPD development(2019) SILVA, Larissa Emidio de Franca; LOURENCO, Juliana Dias; SILVA, Kaique Rodrigues Da; KOHLER, Julia Benini; SANTANA, Fernanda Paula Roncon; MOREIRA, Alyne Riani; CERVILHA, Daniela Aparecida De Brito; HAMAGUCHI, Sara Sumie Sobral; PRADO, Carla Maximo; VIEIRA, Rodolfo De Paula; VELOSA, Ana Paula Pereira; ITO, Juliana Tiyaki; LOPES, Fernanda Degobbi Tenorio Quirino Dos Santos
- Decreased Bone Type I Collagen in the Early Stages of Chronic Obstructive Pulmonary Disease (COPD)(2020) JUNQUEIRA, Jader Joel Machado; LOURENCO, Juliana Dias; SILVA, Kaique Rodrigues da; CERVILHA, Daniela Aparecida de Brito; SILVEIRA, Lizandre Keren Ramos da; CORREIA, Aristides Tadeu; SILVA, Larissa Emidio de Franca; TEODORO, Walcy Rosolia; TIBERIO, Iolanda de Fatima Lopes Calvo; BARBOSA, Alexandre Povoa; LOPES, Fernanda Degobbi Tenorio Quirino dos SantosSmoking is the main risk factor for the development of chronic obstructive pulmonary disease (COPD) and is known to have deleterious effects on bone metabolism. However, the effects on bone collagen matrix during the development of COPD are unclear. The aim of this study was to evaluate the temporal effect of cigarette smoke exposure on bone type I collagen during COPD development in a cigarette smoke-induced model. C57BL/6 mice were allocated to three groups: control (C), animals exposed to filtered air for 1, 3 and 6 months; cigarette smoke (S), animals exposed to cigarette smoke for 1, 3 and 6 months; provisional smoking (PS), animals exposed to cigarette smoke for 3 months, followed by another 3 months of filtered air exposure. Evaluation of the respiratory mechanics and alveolar enlargement were performed. Femoral and tibial extraction was also performed to evaluate the type I collagen by immunofluorescence andCOL1A1gene expression. Exposure to cigarette smoke led to an alveolar enlargement and progressive reduction in lung tissue resistance and elastance, progressive reduction of type I collagen and reduction inCOL1A1gene expression. Although we did not observe any improvement in the functional and histological parameters in the provisional smoking group, we detected an increase inCOL1A1gene expression. A worsening in bone collagen matrix is part of the initial physiopathological events during COPD development and the smoking cessation induced an evident recovery ofCOL1A1expression, possibly to attempt at tissue repair.
- Microenvironmental stimuli induce different macrophage polarizations in experimental models of emphysema(2019) KOHLER, Julia Benini; CERVILHA, Daniela Aparecida de Brito; MOREIRA, Alyne Riani; SANTANA, Fernanda Roncon; FARIAS, Talita M.; VALE, Maria Isabel Cardoso Alonso; MARTINS, Milton de Arruda; PRADO, Carla Maximo; TIBERIO, Iolanda Calvo; ITO, Juliana Tiyaki; LOPES, Fernanda Degobbi Tenorio Quirino dos SantosMacrophages play a pivotal role in the development of emphysema and depending on the microenvironment stimuli can be polarized into M1- or M2-like macrophage phenotypes. We compared macrophage polarizations in cigarette smoke (CS)- and porcine pancreatic elastase (PPE)-induced emphysema models. C57BL/6 mice were subdivided into four experimental groups. In the PPE group, animals received an intranasal instillation of PPE (0.677 IU); in the saline group, animals received an intranasal instillation of saline (0.9%). Animals from both groups were euthanized on day 28. In the CS group, animals were exposed to CS for 30 min, twice a day, 5 days per week for 12 weeks. In the control group, animals received filtered air. We observed an increase in total macrophages for both experimental models. For M1-like macrophage markers, we observed an increase in TNF-alpha(+) and IFN-gamma(+) cells, Cxcl-9 and Cxcl-10 expressions in PPE and CS groups. Only in the CS group, we detected an increased expression of IL-12b. For M2-like macrophages markers we observed a down regulation in IL-10, IL-4, IL-13, Arg1 and Fizz1 and an increase of TGF-beta(+) cells in the PPE group, while for the CS group there was an increase in TGF-beta(+) cells and IL-10 expression. All exposure groups were compared to their respective controls. In summary, we demonstrated that CS- and PPE-induced models resulted in different microenvironmental stimuli. CS exposure induced an environmental stimulus related to M1- and M2-like macrophage phenotypes similar to previous results described in COPD patients, whereas the elastase-induced model provided an environmental stimulus related only to the M1 phenotype.
- Regulatory T cells in COPD development: How the animal model resembles the human pathophysiological features(2017) ITO, Juliana Tiyaki; CERVILHA, Daniela Aparecida de Brito; SILVA, Larissa Emidio de Franca; LOURENCO, Juliana Dias; MOREIRA, Alyne Riani; KOHLER, Julia Benini; NEGRI, Elnara Marcia; MACCHIONE, Mariangela; MAUAD, Thais; MARTINS, Milton Arruda; LOPES, Fernanda Degobbi Tenorio Quirino dos Santos
conferenceObject Th17/Treg Imbalance in Chronic Obstructive Pulmonary Disease (COPD) Development: The Role of Suppressors of Cytokine Signaling (SOCS) and Signal Transducers and Activators of Transcription (STAT) Proteins(2019) SILVA, L. E.; LOURENCO, J. D.; SILVA, K. R.; KOHLER, J. B.; SANTANA, F. P. R.; MOREIRA, A. R.; CERVILHA, D. A. B.; PRADO, C. M.; VELOSA, A. P. P.; VIEIRA, R. P.; ITO, J. T.; LOPES, F. D. Q. S.- A murine model of elastase and cigarette smoke induced emphysema(2017) RODRIGUES, Rubia; OLIVO, Clarice Rosa; LOURENCO, Juliana Dias; RIANE, Alyne; CERVILHA, Daniela Aparecida de Brito; LOPES, Fernanda Degobbi Tenrio Quirino dos SantosObjective: To describe a murine model of emphysema induced by a combination of exposure to cigarette smoke (CS) and instillation of porcine pancreatic elastase (PPE). Methods: A total of 38 C57BL/6 mice were randomly divided into four groups: control (one intranasal instillation of 0.9% saline solution); PPE (two intranasal instillations of PPE); CS (CS exposure for 60 days); and CS + PPE (two intranasal instillations of PPE + CS exposure for 60 days). At the end of the experimental protocol, all animals were anesthetized and tracheostomized for calculation of respiratory mechanics parameters. Subsequently, all animals were euthanized and their lungs were removed for measurement of the mean linear intercept (Lm) and determination of the numbers of cells that were immunoreactive to macrophage (MAC)-2 antigen, matrix metalloproteinase (MMP)-12, and glycosylated 91-kDa glycoprotein (gp91phox) in the distal lung parenchyma and peribronchial region. Results: Although there were no differences among the four groups regarding the respiratory mechanics parameters assessed, there was an increase in the Lm in the CS + PPE group. The numbers of MAC-2-positive cells in the peribronchial region and distal lung parenchyma were higher in the CS + PPE group than in the other groups, as were the numbers of cells that were positive for MMP-12 and gp91phox, although only in the distal lung parenchyma. Conclusions: Our model of emphysema induced by a combination of PPE instillation and CS exposure results in a significant degree of parenchymal destruction in a shorter time frame than that employed in other models of CS-induced emphysema, reinforcing the importance of protease-antiprotease imbalance and oxidant-antioxidant imbalance in the pathogenesis of emphysema.
- Th17/Treg imbalance in COPD progression: A temporal analysis using a CS-induced model(2019) ITO, Juliana Tiyaki; CERVILHA, Daniela Aparecida de Brito; LOURENCO, Juliana Dias; GONCALVES, Natalia Gomes; VOLPINI, Rildo Aparecido; CALDINI, Elia Garcia; LANDMAN, Gilles; LIN, Chin Jia; VELOSA, Ana Paula Pereira; TEODORO, Walcy Paganelli Rosolia; TIBERIO, Iolanda de Fatima Lopes Calvo; MAUAD, Thais; MARTINS, Milton de Arruda; MACCHIONE, Mariangela; LOPES, Fernanda Degobbi Tenorio Quirino dos SantosBackground The imbalance between pro- and anti-inflammatory immune responses plays a pivotal role in chronic obstructive pulmonary disease (COPD) development and progression. To clarify the pathophysiological mechanisms of this disease, we performed a temporal analysis of immune response-mediated inflammatory progression in a cigarette smoke (CS)-induced mouse model with a focus on the balance between Th17 and Treg responses. Methods C57BL/6 mice were exposed to CS for 1, 3 or 6 months to induce COPD, and the control groups were maintained under filtered air conditions for the same time intervals. We then performed functional (respiratory mechanics) and structural (alveolar enlargement) analyses. We also quantified the NF-kappa B, TNF-alpha, CD4, CD8, CD20, IL-17, IL-6, FOXP3, IL-10, or TGF-beta positive cells in peribronchovascular areas and assessed FOXP3 and IL-10 expression through double-label immunofluorescence. Additionally, we evaluated the gene expression of NF-kappa B and TNF in bronchiolar epithelial cells. Results Our CS-induced COPD model exhibited an increased proinflammatory immune response (increased expression of the NF-kappa B, TNF-alpha, CD4, CD8, CD20, IL-17, and IL-6 markers) with a concomitantly decreased anti-inflammatory immune response (FOXP3, IL-10, and TGF-beta markers) compared with the control mice. These changes in the immune responses were associated with increased alveolar enlargement and impaired lung function starting on the first month and third month of CS exposure, respectively, compared with the control mice. Conclusion Our results showed that the microenvironmental stimuli produced by the release of cyto-kines during COPD progression lead to a Th17/Treg imbalance.
- Regulatory T-Cell Distribution within Lung Compartments in COPD(2017) SALES, Davi S.; ITO, Juliana T.; ZANCHETTA, Ivy A.; ANNONI, Raquel; AUN, Marcelo V.; FERRAZ, Luiz Fernando S.; CERVILHA, Daniela A. B.; NEGRI, Elnara; MAUAD, Thais; MARTINS, Milton A.; LOPES, Fernanda D. T. Q. S.The importance of the adaptive immune response, specifically the role of regulatory T (Treg) cells in controlling the obstruction progression in smokers, has been highlighted. To quantify the adaptive immune cells in different lung compartments, we used lung tissues from 21 never-smokers without lung disease, 22 current and/or ex-smokers without lung disease (NOS) and 13 current and/or ex-smokers with chronic obstructive pulmonary disease (COPD) for histological analysis. We observed increased T, B, IL-17 and BAFF(+) cells in small and large airways of COPD individuals; however, in the NOS, we only observed increase in T and IL-17(+) cells only in small airways. A decrease in the density of Treg(+), TGF-beta(+) and IL-10(+) in small and large airways was observed only in COPD individuals. In the lymphoid tissues, Treg, T,B-cells and BAFF(+) cells were also increased in COPD; however, changes in Treg inhibitory associated cytokineswere not observed in this compartment. Therefore, our results suggest that difference in Treg(+) cell distributions in lung compartments and the decrease in TGF-beta(+) and IL-10(+) cells in the airways may lead to the obstruction in smokers.