Banca de DEFESA: MARIANA BORGES LOPES

Uma banca de DEFESA de DOUTORADO foi cadastrada pelo programa.
STUDENT : MARIANA BORGES LOPES
DATE: 29/09/2020
TIME: 13:00
LOCAL: Videoconferência
TITLE:

miRNA-mRNA Integrative Analysis of Diabetes-induced Cardiomyopathy in Rats

KEY WORDS:

Diabetic cardiomyopathy; mimic miRNAs; rno-miR-214-3p; Pla2g2a; Il6; gene therapy.

PAGES: 144
BIG AREA: Ciências da Saúde
AREA: Farmácia
SUMMARY:

Systemic hyperglycemia caused by diabetes leads to an inflammatory process and cardiovascular dysfunctions including diabetic cardiomyopathy (DCM). The present study aimed to search through in silico analyzes, mRNAs and miRNAs differently expressed in DCM and, in vitro and in vivo, to validate the role of rno-miR-214-3p through the regulation of phospholipase A2 of group IIA (Pla2g2a) in inflammatory process resulting from hyperglycemia. Thus, the expression of rno-miR-214-3p, Pla2g2a and Il6, was avaliated in Wistar rats with streptozotocin-induced diabetes (40 mg / kg, iv) left ventricle (LV) and in H9c2 cell culture under normoglycemic conditions (NG , 5.5 mmol / L glucose), hyperglycemic (25 mmol / L glucose) and transfected with rno-miR-214-3p mimetic in hyperglycemic medium to validate the in silico data. Total RNA and proteins were extracted from the samples and gene expression was evaluated by RT-qPCR and Western Blotting. Fibrosis was characterized by increased collagen deposition in interstitial space in the myocardium of diabetic rats (p=0.023). Hyperglycemic conditions resulted in downregulation of rno-miR-214-3p (p=0.043 and p=0.020) and upregulation of Pla2g2a (p=0.016 and p=0.043) and Il6 (p=0.016 and p=0.011) in rat LV and H9c2 cells, respectively. Protein expression of sPLA2-IIA (p=0.011) and IL-6 (p=0.011) was also increased in LV of diabetic rats. mRNA and protein expression of Pla2g2a correlated positively with glycaemia and Il6 expression (p<0.05). Transfection of H9c2 cells with mimic increased rno-miR-214-3p and reduced Pla2g2a and Il6 expression under HG concentrations (50 nM for 24 h). The present study confirmed our in silico results in which, hyperglycemia leads to a reduction in the expression of rno-miR-214-3p and an increase in its target gene Pla2g2a. The increase in rno-miR-214-3p expression reduced the expression of Pla2g2a and Il6, indicating that miR-214 / Pla2g2a / Il6 may be a pathway of inflammation associated with the development of DCM.

BANKING MEMBERS:
Externa à Instituição - ALICE CRISTINA RODRIGUES - USP
Externo à Instituição - ALVARO DANILO CERDA MAUREIRA - UFRO
Presidente - 2087759 - ANDRE DUCATI LUCHESSI
Interno - 2275890 - MARCELO DE SOUSA DA SILVA
Interno - 1544647 - MATHEUS DE FREITAS FERNANDES PEDROSA