Expression of sPLA2-IIA protein in the context of inflammation in diabetic cardiomyopathy

• Autor(a) principal: Abreu, Lucas Andriel Carvalho
• Data de Publicação: 2019
• Tipo de documento: Trabalho de conclusão de curso
• Idioma: por
• Título da fonte: Repositório Institucional da UFRN
• Texto Completo: https://repositorio.ufrn.br/handle/123456789/35767
• topic: Diabetes mellitus; H9c2 cell line; animal model; interleukin-6; group IIA phospholipase 2

Diabetic cardiomyopathy (DCM) is a chronic complication of diabetes, associated with high mortality and morbidity in diabetic patients. In this context, the present study sought to relate the gene and protein expression of phospholipase 2 of group IIA and inflammatory interleukin 6 cytokines involved in the development of CMD through in vitro and in vivo approaches. In the in vitro study, culture of the H9c2 myoblast lineage was performed under normoglycemic (NG, 5.5 mmol / L glucose) and hyperglycemic (HG, 25 mmol / L glucose) conditions, total RNA from cells was extracted for analysis of gene expression of (IL-6) - Interleukin 6 and (sPLA2-IIA) - Group II A secretory phospholipase A2 (protein) by real time PCR (RTqPCR). In the in vivo model, Wistar rats had pharmacologically streptozotocin-induced diabetes (STZ) (40 mg / kg, iv; n = 7) and were compared to a control group (n = 7), thirtes days after induction, the animals were euthanized and their left ventricles used for IL-6 and sPLA2-IIA protein extraction. Pla2g2a gene expression was increased in H9c2 culture under hyperglycemic conditions (3-fold increased, p = 0.043). Inflammatory cytokine IL-6 was increased in in vitro expression analysis (p = 0.0095). For Western blot protein expression analysis, paraffin blocks containing the left ventricle of diabetic rats were compared to a control group. It was observed that sPLA2-IIA expression was approximately six-fold higher in the diabetic rat when compared to the control (p <0.001). Not so different, IL-6 was also doubled in expression (p = 0.016), corroborating previous results of gene expression in cell culture. The increased expression of these molecules when in hyperglycemic environment may be correlated with the inflammatory process resulting from chronic hyperglycemia that would be a possible pathway for the development of cellular changes observed in DCM, thus becoming a possible marker of development of this complication in diabetic individuals.