Influence of physical exercise on cardiac remodeling, oxidative stress, MAPK and NF-kB pathways signaling in spontaneously hypertensive rats

Author(s)

Luana Urbano Pagan

• Document type: Doctoral Thesis
• Press: Botucatu. 2018-03-16.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Medicina. Botucatu
• Defense date: 2018-03-02
• Advisor: Katashi Okoshi

Abstract

Introduction: The pressure overload caused by systemic arterial hypertension (SAH) may change the collagen architecture, induce fibrosis, as well as imbalance between the reactive oxygen species (ROS) production and antioxidant capacity. Increased ROS leads to activation of signaling pathways such as nuclear factor kappa B (NF-kB) and mitogen-activated protein kinases (MAPK). Alterations in these pathways contribute to cardiac remodeling process induced by SAH. Physical exercise plays an important role in mitigating cardiovascular risk factors such as hypertension. Therefore, the aim of this study was to evaluate the influence of physical training, started before clinical evidence of heart failure, on cardiac remodeling in spontaneously hypertensive rats (SHR). Methods: Four experimental groups were used: sedentary (W-SED n=27) and trained (W-EX, n=31) normotensive Wistar rats, and sedentary (SHR-SED, n=27) and exercised (SHR-EX, n=32) hypertensive rats. Rats of the exercise groups underwent a protocol of treadmill exercise five days a week, for four months; exercise started at 13 months of age. Echocardiogram was performed to evaluate in vivo cardiac structures and function. In vitro myocardial function was analyzed in left ventricular (LV) papillary muscle preparations. LV tissue samples were obtained for biochemical, histological, and molecular analysis. Total myocardial collagen was assessed by histology and hydroxyproline quantification. Cardiomyocyte size was measured in LV histological sections. Antioxidant enzymes activity was quantified by spectrophotometry. NADPH oxidase activity was analyzed by reduction of lucigenin. Protein expression of collagen I and III, lysyl oxidase, MAPK and NF-kB, and metalloproteinases tissue inhibitors 1 and 2 was quantified by Western blot. The activity of metalloproteinases was evaluated by zymography. Comparisons between groups were performed by two factors analysis of variance (ANOVA), complemented with the Bonferroni test (normal distribution), or Kruskal-Wallis complemented with Dunn test (non-normal distribution). Results: Systolic blood pressure was higher in the SHR groups. The exercised groups showed greater physical capacity. Prevalence of heart failure signs was higher in the hypertensive groups compared to controls, and the SHR-EX group showed lower prevalence of pleural effusion and tachypnea compared to SHR-SED. Echocardiogram showed lower LV wall thickness, LV relative wall thickness, left atrium diameter, and relaxation time in the SHR-EX group vs. SHR-SED. Myocardial functional study showed better performance in the SHR-EX group (positive derivative of the developed tension) vs. SHR-SED. The SHR-EX group showed higher antioxidant enzymes activity compared to SHR-SED. Lipid hydroperoxide production, myocyte diameters, and phosphorylated JNK and total IkB protein expression were higher in the hypertensive groups. Quantification of hydroxyproline, malondialdehyde, NADPH oxidase activity, and protein expression of collagen III, lysyl oxidase, TIMP-1, total JNK, phosphorylated p38, phosphorylated and total p65, and phosphorylated IkB did not differ between groups. The interstitial collagen fraction, MMP-2 activity, protein expression of total p38, and total and phosphorylated ERK were higher in the SHR-SED group compared to normotensive control. Physical exercise reduced the MMP-2 activity and the phosphorylated ERK expression in hypertensive rats. Conclusion: Physical exercise in spontaneously hypertensive rats attenuates cardiac remodeling associated with improved physical capacity and reduced prevalence of heart failure signs. In addition, it is associated with increased antioxidant enzymes activity, decreased ERK phosphorylation and MMP-2 activity, and attenuation of total ERK protein expression. (AU)

• FAPESP's process: 14/00747-1 - Influence of physical exercise on cardiac remodeling, oxidative stress, MAPK and NF-kB pathways signaling in spontaneously hypertensive rats
• Grantee: Luana Urbano Pagan
• Support Opportunities: Scholarships in Brazil - Doctorate