Análise do mecanismo de ação do Alfa-Terpineol em células de músculo liso A7r5

Abstract

Alpha-Terpineol is a promising monoterpene for studies related to hypertension due to its action in reducing blood pressure in hypertensive rats (SABINO et al., 2013). The present work aims to analyze the possible mechanisms of action of Alpha-Terpineol in A7r5 smooth muscle cells that may explain the hypotensive action of this monoterpene observed in vivo. We used cell culture, cell viability analysis and Alpha-Terpineol cytotoxicity using the MTT assay, RNA extraction and cDNA synthesis. We used the MTT cell viability test at 24, 48 and 72 hours of incubation. Results demonstrate that in the cytotoxicity tests, alpha-terpineol promoted cell death only at the highest concentration of 10 -2 M, in the periods of 24 and 48 hours. Within 72 hours there was a reduction in cell viability at concentrations of 10 -5 and 10 -3 M and cell death at concentration of 10 -2 M. To investigate possible changes in mRNA of adenosine A2A receptors, mRNA expression was evaluated by real-time PCR. A statistically significant increase in mRNA expression of adenosine A2A receptors in A7r5 cells was observed following 6 hour-treatment with Alpha-Terpineol concentration of 10 -8 M. Using in Cell Western Blotting assay the expression of A2a receptor was significantly reduced by Alpha-Terpineol at all periods tested. Our data show that Alpha-Terpineol action on smooth muscle cells may involve adenosine A2A receptors and open new perspectives for studying its mechanism of action in these cells.