Evaluation of atorvastatin pleiotropic effects on the regulation of epigenetics mechanisms over specific gens linked to nitric oxide on a hypertension experimental model

Abstract

Nitric oxide (NO) is a potent vasodilator, being responsible for maintaining a healthy vascular endothelium. Nitric oxide synthases (NOS) of the endothelial type (eNOS - NOSIII) control blood pressure and elicit protecting effects on vessels. Statins cause effects that go beyond the classical mechanism (pleiotropic effects), among them increasing the expression of eNOS and consequently increasing NO synthesis. Among statins, atorvastatin has been shown to induce global epigenetic modifications in cancer. Objective: To assess whether atorvastatin elicit pleiotropic effects through regulation of specific genes involved in NO synthesis in a renovascular hypertension model. Specific objectives: A) to assess the time-course relationship between hypertension and the epigenetic regulation of NOS3 in endothelial cells and smooth muscle cells of rats. B) To assess whether the dose-dependent effects of Atorvastatin change the regulation of NOS3 methylation in hypertensive or sham operated rats in the presence or absence of a previous treatment with decitabine (a known demethylating agent). Materials and Methods: Two experimental protocols will be performed: a time course of the disease model, and an intervention with atorvastatin in sham or 2K1C renovascular hypertension rats with or without a previous treatment with decitabine. We will use heterogenic Wistar Hannover rats, and after experimental protocols, rats will be euthanized and samples of blood and aorta will be harvested. Aorta reactivity assays, nitrite determination, DNA methyl transferase 1 assay, assessment of CpG islands in NOS3 gene and expression of eNOS through qPCR and western blot will be assessed in samples. Results will be analyzed by ANOVA or Kruskal-Wallis if parametric or non-parametric, respectively. (AU)