Banca de QUALIFICAÇÃO: ANA CAROLINA RIBEIRO OLIVEIRA E SILVA

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
DISCENTE : ANA CAROLINA RIBEIRO OLIVEIRA E SILVA
DATA : 20/06/2022
HORA: 09:00
LOCAL: Campus Santo André
TÍTULO:

Evaluation of Cyp3a13, member of cytochome P450 superfamily, Role in Non-Alcoholic Fatty Liver Disease (NAFLD) Development

PÁGINAS: 52
GRANDE ÁREA: Ciências Biológicas
ÁREA: Bioquímica
SUBÁREA: Metabolismo e Bioenergética
RESUMO:

Non-alcoholic fatty liver disease (NAFLD) is considered the hepatic manifestation of metabolic syndrome and has reached alarming rates, being considered the most common liver disorder on the planet. Characterized by hepatic steatosis (HE), NAFLD in its first stage is benign and reversible. However, as it is asymptomatic, it can progress to severe inflammatory conditions such as steatohepatitis, cirrhosis and hepatocarcinoma, which leads to impairment of the vital functions of the liver. NAFLD is a multifactorial disease, being associated with insulin resistance, type 2 diabetes mellitus (DM2), obesity, dyslipidemia and intestinal microbiota. In the present study, we built a lentiviral tool controlled by the Tight inducible promoter in order to investigate the role of the CYP3A13 enzyme, a member of the cytochrome p450 superfamily (CYP450), in AML-12 cells previously expressing the TET3G system, under steatosis conditions. Objective: To analyze the possible role of CYP3A13 in the development of HE and its metabolic alterations, through overexpression by the inducible TetOn system, in AML-12 lineage hepatocytes. Methods: A lentiviral tool controlled by the Tight inducible promoter was constructed carrying the molecular marker GFP and expressing the murine gene CYP3A13, under doxycycline stimulation. AML-12 cells previously expressing the 3G protein of the TetOn system received pLENTI Tight CYP3A13 IRES eGFP by lipofectamine assay, and were selected for 11 days with geneticin and blasticidin. Assays mimicking dyslipidemia were performed. Changes in gene expression were analyzed by RT-qPCR and the cells stained with Oil Red O (ORO) to verify the accumulation of neutral triglycerides. Results: Lipid accumulation can be assessed through reesterification of fatty acids by staining with ORO. Overexpression of the CYP3A13 enzyme has no effect on oleate metabolism. Conclusion: The tool built is functional for studies involving the CYP3A13 enzyme, however other metabolic pathways need investigation.

MEMBROS DA BANCA:
Presidente - Interno ao Programa - 1227329 - CESAR AUGUSTO JOAO RIBEIRO
Membro Titular - Examinador(a) Interno ao Programa - 1672728 - ANA CAROLINA SANTOS DE SOUZA GALVAO
Membro Titular - Examinador(a) Externo à Instituição - MONICA MARQUES TELLES - UNIFESP
Membro Suplente - Examinador(a) Interno ao Programa - 1674592 - TIAGO RODRIGUES
Membro Suplente - Examinador(a) Externo à Instituição - ANAYSA PAOLA BOLIN - USP