FIBROGÊNESE PULMONAR NA PARACOCCIDIOIDOMICOSE: COMPARAÇÃO DO PERFIL PROTÊOMICO DE DIFERENTES MODELOS EXPERIMENTAIS DE FIBROSES PULMONAR E HEPÁTICA .

• Ano de defesa: 2022
• Autor(a) principal: Amanda Ribeiro dos Santos
• Orientador(a): James Venturini
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Fundação Universidade Federal de Mato Grosso do Sul
• País: Brasil
• Palavras-chave em Português: Paracoccidioides brasiliensis, fibrose pulmonar, enfisema, proteômica, fibrose hepática, paracoccidioidomicose.
• Link de acesso: https://repositorio.ufms.br/handle/123456789/5393

Introduction. Paracoccidioidomycosis (PCM) is a systemic mycosis caused by fungi of the genus Paracoccidioides. It is an endemic disease in Latin America, being the leading cause of death among all systemic mycoses in Brazil. This disease mainly affects rural workers and induces severe sequelae, including pulmonary fibrosis (PF) and pulmonary emphysema. In general, fibrogenesis is characterized by myofibroblast hyperplasia and intense collagen deposition in the parenchyma and blood vessels. Over time, this process induces changes in the architecture of the affected organ, inducing a decline in function. In general, knowledge of the mechanisms involved in fibrogenesis is based on studies, for example, of non-infectious hepatic and pulmonary fibrosis, such as idiopathic pulmonary fibrosis (IPF). In PCM, the mechanisms involved in PF remain unclear. Objective. Identify signaling pathways relevant to FP in experimental PCM by comparing the pulmonary proteome of mice infected with P. brasiliensis and mice with bleomycin-induced FP (BLM), as well as relevant target proteins that are common in FP models (PCM and BLM) and in the model of FH induced by carbon tetrachloride (CCl4). Material and Methods. The FP-PCM model consisted of adult male BALB/c mice inoculated with P. brasiliensis yeasts (Pb326 isolate) by the intratracheal route and evaluated after eight weeks. The FP-BLM model consisted of adult male BALB/c mice, which were administered three doses of bleomycin intraperitoneally and were evaluated two weeks after the last dose. The FH model consisted of adult male C57Bl/6 mice in which 12 doses of CCl4 were administered intraperitoneally twice a week for six weeks. Control groups consisted of BALB/c and C57Bl/6 mice submitted to the same inoculum conditions, using sterile saline solution. Lungs and liver were collected according to each model and submitted to histopathological analysis and recovery of viable fungi for the FP-PCM model. Proteomic analysis was performed using liquid nanochromatography coupled to an electrospray ionization mass spectrophotometer (nano-LC-ESI MS/MS). The Protein Lynx Global Service (PLGS) software was used to identify the difference in the expression of the identified proteins, where p <0.05 and 1 - p> 0.95, were used for the determination of under- or over expressed proteins respectively. Bioinformatics analyzes were performed to identify the significantly enriched pathways in which proteins with similar or different expression between groups participated. For this analysis, the Reactome pathway database was used through the Cluego v2.0.7 + Clupedia v1.0.8 plug-in in the Cytoscape software. Results and discussion. Proteomic analysis revealed 919 proteins differentially expressed between the lungs of the FP-PCM model and the healthy lung of the control group. It was also observed that P. brasiliensis infection induced overexpression in pathways related to pro-fibrotic signaling, including: neutrophilic response, cellular response to stress, pro-fibrotic response mediated by TGF-β receptor signaling in cell transition from epithelial to mesenchymal, attenuation phase and secretory phenotype associated with senescence, platelet degranulation and PI3K signaling. A total of 355 proteins were identified in common with the three fibrosis models evaluated. The analysis of the expression pattern of these proteins through heatmap revealed that the fibrogenesis observed in the experimental PCM showed greater similarity between the proteins differentially expressed by the model of FH induced by CCl4. Rho GTPases, HSP-90 and vimentin proteins were overexpressed in the fibrotic tissues of these two groups. Conclusion. Our findings contribute to identify more specific molecular mechanisms involved in PCM fibrogenesis, as well as to identify protein targets for possible anti-fibrotic drugs in PCM