DIOL Triterpenes Block Profibrotic Effects of Angiotensin II and Protect from Cardiac Hypertrophy

et al. (2012) DIOL Triterpenes Block Profibrotic Effects of Angiotensin II and Protect from Cardiac Hypertrophy. PLoS ONE 7(7): e41545. doi:10.1371/journal.pone.0041545 DIOL Triterpenes Block Profibrotic Effects of Angiotensin II and Protect from Cardiac Hypertrophy Ruben Martn 0 Maria Miana 0 Raquel Jurado-Lo pez 0 Ernesto Martnez-Martnez 0 Nieves Go mez- 0 Hurtado 0 Carmen Delgado 0 Maria Visitacio n Bartolome 0 Jose Alberto San Roma n 0 Claudia Cordova 0 Vicente Lahera 0 Maria Luisa Nieto 0 Victoria Cachofeiro 0 Luis Eduardo M. Quintas, Universidade Federal do Rio de Janeiro, Brazil 0 1 Departamento de Fisiolog a, Facultad de Medicina, Universidad Complutense , Madrid , Spain , 2 Instituto de Ciencias del Coraz o n (ICICOR), Hospital Cl nico , Valladolid , Spain , 3 Departamento de Farmacolog a, Facultad de Medicina. Universidad Complutense , Madrid , Spain , 4 Centro de Investigaciones Biol o gicas, Consejo Superior de Investigaciones Cient ficas (CSIC) , Madrid , Spain , 5 Departamento de Oftalmolog a y Otorrinolaringolog a, Facultad de Psicolog a, Universidad Complutense , Madrid, Spain, 6 Instituto Biolog a y Gene tica Molecular, CSIC-UVA, Valladolid , Spain Background: The natural triterpenes, erythrodiol and uvaol, exert anti-inflammatory, vasorelaxing and anti-proliferative effects. Angiotensin II is a well-known profibrotic and proliferative agent that participates in the cardiac remodeling associated with different pathological situations through the stimulation and proliferation of cardiac fibroblasts. Therefore, the aim of the study was to investigate the preventive effects of the natural triterpenes erythrodiol and uvaol on the proliferation and collagen production induced by angiotensin II in cardiac myofibroblasts. Their actions on cardiac hypertrophy triggered by angiotensin II were also studied. Methodology/Principal Findings: The effect of erythrodiol and uvaol on angiotensin II-induced proliferation was evaluated in cardiac myofibroblasts from adult rats in the presence or the absence of the inhibitors of PPAR-c, GW9662 or JNK, SP600125. The effect on collagen levels induced by angiotensin II was evaluated in cardiac myofibroblasts and mouse heart. The presence of low doses of both triterpenes reduced the proliferation of cardiac myofibroblasts induced by angiotensin II. Pretreatment with GW9662 reversed the effect elicited by both triterpenes while SP600125 did not modify it. Both triterpenes at high doses produced an increase in annexing-V binding in the presence or absence of angiotensin II, which was reduced by either SP600125 or GW9662. Erythrodiol and uvaol decreased collagen I and galectin 3 levels induced by angiotensin II in cardiac myofribroblasts. Finally, cardiac hypertrophy, ventricular remodeling, fibrosis, and increases in myocyte area and brain natriuretic peptide levels observed in angiotensin II-infused mice were reduced in triterpene-treated animals. Conclusions/Significance: Erythrodiol and uvaol reduce cardiac hypertrophy and left ventricle remodeling induced by angiotensin II in mice by diminishing fibrosis and myocyte area. They also modulate growth and survival of cardiac myofibroblasts. They inhibit the angiotensin II-induced proliferation in a PPAR-c-dependent manner, while at high doses they activate pathways of programmed cell death that are dependent on JNK and PPAR-c. - Funding: This work was supported by grants from Fondo de Investigaciones Sanitarias (PI09/0871). and Junta de Castilla y Leo n (CSI11A08). Maria Miana and Raquel Jurado-Lo pez were paid with a Grant from Red Cardiovascular del FIS (RD06/0014/0007). Ruben Martin was paid with a Grant from La Fundacio n General de la Universidad de Valladolid (FGUVa) (060/053186). Claudia Cordova was funded by the FPI Program from the Government of Castilla y Leo n (co-funded by FSE). All the authors are members of the Red Cardiovascular del FIS (RECAVA, RD06/0014/0007 and RD06/0014/0000). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. . These authors contributed equally to this work. "These authors also contributed equally to this work. Cardiac fibroblasts are one of the major cellular components of the heart. They play an important role in the maintenance of structural integrity and normal cardiac function, where both cellcell and cell-extracellular matrix interactions are essential [1,2]. They participate in the reparative response of damaged tissue to wound healing, not only through controlled extracellular matrix production, but also through proliferation, migration and differentiation into hypersecretory myofibroblasts [35]. The acquisition of smooth-muscle-like properties in fibroblasts is associated with exacerbation of extracellular matrix production [6], which can trigger impairment of cardiac function by facilitating reduced contractibility and arrhythmias, and which then ultimately contribute to heart failure [79]. The activation of cardiac fibroblasts to myofibroblasts is greatly enhanced in chronic cardiac diseases and after acute cardiac events [911]. This transformation is controlled by a variety of stimuli, including growth and vasoactive factors such as angiotensin II, cytokines and mechanical stimuli [12]. Angiotensin II plays a central role in the development and complications of cardiovascular diseases by exerting, among other types of action, a fibrotic one [1315]. This participation has been demonstrated by the effectiveness of drugs that interact with this system on patients with left ventricular hypertrophy or heart failure [15]. Its fibrotic action involves the activation not only of growth factors such as connective tissue growth factor (CTGF) but also new mediators such as galectin 3, which is associated with adverse long-term cardiovascular outcomes in patient with heart failure [16,17]. The Mediterranean diet, in which olive oil is the major source of dietary fat intake, has been associated with low incidence of cardiovascular diseases [18,19] and cancer [2022]. Although these health benefits have long been attributed to a high content of monounsaturated fatty acids (oleic acid), a wide variety of minor components are under evaluation. Among these bioactive compounds are the triterpenes including the diols, uvaol and erythrodiol [23]. Many pharmacological properties, including antiinflammatory, antitumoral and antioxidant activities [2426], have been reported for these compounds. In addition, recent studies have suggested beneficial effects on the cardiovascular system, since antihypertensive vasodepressor, cardiotonic, and antidysrhythmic properties have been reported [2729]. However, the effect of these compounds on normal cells, especially on cardiac cells, is unknown. Thus, in the search for novel pharmacological approaches for the management of cardiovascular pathologi (...truncated)