Banca de QUALIFICAÇÃO: GRAZIELE CRISTINA FERREIRA

Uma banca de QUALIFICAÇÃO de DOUTORADO foi cadastrada pelo programa.
DISCENTE : GRAZIELE CRISTINA FERREIRA
DATA : 03/09/2019
HORA: 14:00
LOCAL: sala S24, Térreo, Bloco Delta, Campus SBC da Fundação Universidade Federal do ABC, localizada na Alameda da Universidade, s/n, Bairro Anchieta - São Bernardo do Campo, SP
TÍTULO:

Analysis of the serine protease inhibitor (rBmTI-A) effect on pulmonary A549 cells

PÁGINAS: 50
GRANDE ÁREA: Ciências Biológicas
ÁREA: Bioquímica
SUBÁREA: Biologia Molecular
RESUMO:

         The pumonary emphysema is characterized by the alveolar dammage, and is one of COPD (Chronic obstructive pulmonary disease) manifestation. COPD is one of the leading causes of mortality in the world. Some proteases are envolved with the emphysema development process, such as human neutrophil elastase (HNE).

            The serine protease inhibitor rBmTI-A is originary from Rhipicephalus Boophilus microplus, and belongs to Kunitz-BPTI inhibitor Family. rBmTI-A has two inhibitory domains and activity on HNE, trypsin, plamin and Human Plasma Kallikrein. In a previous work of our team we demonstred that rBmTI-A is able to prevent the pulmonary emphysema development, induced by porcine pancreatic elastase (PPE), in mice. Apparently this effect is due the inflammatory  response control by rBmTI-A.

            In the presente work, it was analysed the inflammatory response induced by PPE in human pulmonary cells A549 and the effect of rBmTI-A in these cells. The rBmTI-A was expressed, purified and biochemical analyses were performed to test the activity of the recombinant inhibitor.  The A549 cells were incubated in different PPE concentrations for 1 hour and 16 hours, both conditions with control without PPE. The culture supernatant and the cells to evaluate the detachment level, cellular viability, proteolysis analysis by culture supernatant and quantitative real time PCR of IL8 and IL6. Also it was performed assays using rBmTI-A in different concentrations and PPE (0.0125 U/mL). In this assays rBmTI-A was incubated with cells for two hours followed by PPE add and another incubation were done for 16h  , as read-outs we collect data of cell viability and ELISA for IL8.

            The results demonstrated that in both incubation protocols  of A549 cells (1h and 16h) with PPE IL8 and IL6 were expressed with significant diferences compared with controls.

            In the assay using rBmTI-A and PPE the concentration of 28 nM presented the best result, 71% of IL-8 was reduced compared with control only with PPE, no interferece in cellular viability were observed in combined assay (rBmTI-A and PPE) compared with only PPE. We concluded that PPE is able to induce IL6 and IL-8 secretion in A549 cells in concentrations that no interfere in cell viability, and that rBmTI-A is able to avoid this IL-8 secretion, probably interfering in the induction of inflammatory response in A549 cells by PPE.

MEMBROS DA BANCA:
Presidente - Interno ao Programa - 1672728 - ANA CAROLINA SANTOS DE SOUZA GALVAO
Membro Titular - Examinador(a) Externo ao Programa - 3066269 - VINICIUS DE ANDRADE OLIVEIRA
Membro Titular - Examinador(a) Externo ao Programa - 1941387 - FERNANDA DIAS DA SILVA
Membro Suplente - Examinador(a) Interno ao Programa - 2605490 - SERGIO DAISHI SASAKI