Preventive Effect of Yuzu and Hesperidin on Left Ventricular Remodeling and Dysfunction in Rat Permanent Left Anterior Descending Coronary Artery Occlusion Model

Jung Y-S (2015) Preventive Effect of Yuzu and Hesperidin on Left Ventricular Remodeling and Dysfunction in Rat Permanent Left Anterior Descending Coronary Artery Occlusion Model. PLoS ONE 10(1): e110596. doi:10.1371/journal.pone.0110596 Preventive Effect of Yuzu and Hesperidin on Left Ventricular Remodeling and Dysfunction in Rat Permanent Left Anterior Descending Coronary Artery Occlusion Model Hye Yon Yu 0 Ji Hun Ahn 0 Se Won Park 0 Yi-Sook Jung 0 Vincenzo Lionetti, Scuola Superiore Sant'Anna, Italy 0 1 Department of Physiology, School of Medicine, Ajou University , Suwon , Republic of Korea, 2 College of Pharmacy, Ajou University , Suwon , Republic of Korea, 3 Department of Cardiology, Soon Chun Hyang University Gumi Hospital , Gumi , Republic of Korea, 4 Molecular Biotechnology , College of Life and Environmental Sciences, Konkuk University , Seoul , Republic of Korea, 5 Research Institute of Pharmaceutical Sciences and Technology, Ajou University , Suwon , Republic of Korea Left ventricular (LV) remodeling, which includes ventricular dilatation and increased interstitial fibrosis after myocardial infarction (MI), is the critical process underlying the progression to heart failure. Therefore, a novel approach for preventing LV remodeling after MI is highly desirable. Yuzu is a citrus plant originating in East Asia, and has a number of cardioprotective properties such as hesperidin. However, no study has proved whether yuzu can prevent LV remodeling. The aim of this study was to determine the effects of yuzu on heart failure (HF) and its potential impact on the LV remodeling process after MI. Our in vivo study using the permanent left anterior descending coronary artery (LAD) occlusion model demonstrate that one week pre-treatment with yuzu or its major metabolite hesperidin before LAD occlusion significantly attenuated cardiac dysfunction, myocyte apoptosis and inflammation. Not only yuzu but also hesperidin inhibited caspase-3 activity, myeloperoxidase expression, a-smooth muscle actin expression, and matrix metalloproteinase2 activity in a permanent LAD occlusion rat model. To our knowledge, our findings provide the first evidence that yuzu and hesperidin prevent MI-induced ventricular dysfunction and structural remodeling of myocardium. - LV remodeling is pathologic changes in the architecture of the LV that occur due to various cardiovascular diseases (CVDs) including MI and hypertension [1]. Of these, MI is caused by the partial interruption or occlusion of the blood supply to a part of the myocardium. This most commonly involves the occlusion of a coronary artery following the rupture of a vulnerable atherosclerotic plaque [2]. LV remodeling after MI is associated with a combination of pathologic conditions, including myocyte hypertrophy, myocyte apoptosis, myofibroblast proliferation, inflammatory reaction, and interstitial fibrosis, which ultimately lead to the loss of systolic and diastolic function [3]. Cardiac hypertrophy is a compensatory process in response to increased hemodynamic overload, characterized by an increase in the size of individual cardiac myocytes and wall thickness. On the other hand, in chronic MI following LAD occlusion, a transition occurs from compensatory cardiac hypertrophy to decompensatory hypertrophy, characterized by a chamber dilation and wall thinning. In this chronic condition, processes such as extracellular matrix turnover, fibrosis, inflammation and apoptosis are crucial determinants [4,5]. LV remodeling after MI is a key contributor to HF, which is one of the most common causes of cardiovascular morbidity and mortality worldwide [6]. Conventional HF therapy is still largely based on targeting the causes and neurohumoral activation of HF, and includes agents such as angiotensin-converting enzyme inhibitors, angiotensin-receptor antagonists, beta-blockers, and aldosterone antagonist [7]. Recently, natural products have become popular worldwide and have gained wide acceptance as adjuncts to conventional therapy. Various studies have shown natural products such as grapes, citrus fruits, broccoli, and cacao are rich sources of phytochemicals such as polyphenols that are well known for their antioxidant and cardioprotective effects [8,9,10]. Furthermore, epidemiological evidence indicates that a negative correlation exists between the consumption of flavonoidrich foods and the incidence of CVDs [11]. Yuzu (Citrus junos Sieb ex Tanaka), one of the natural products receiving attention for their health benefits, is a citrus fruit native to northeast Asia, including Korea, China, and Japan. It has been used in traditional medicine in northeast Asia, and it is known to improve blood circulation and prevent colds [12]. We have previously reported that yuzu and its major compounds inhibit platelet aggregation in vivo and in vitro [12]. However, little report has described whether yuzu has beneficial effect against cardiac dysfunction following chronic MI. The goal of the present study was to evaluate the effects of yuzu in a rat model of LV remodeling induced by permanent LAD occlusion. Considering that hesperidin is well-known major functional component of yuzu, we have also evaluated whether hesperidin contributes to the protective effect of yuzu. Materials and Methods Materials Ethanolic extract of yuzu were obtained from Konkuk University (Seoul, Republic of Korea). Briefly, Yuzu fruits minced were extracted with ethanol and lyophilized to remove solvent. Yuzu extract was dissolved in saline (0.9% NaCl) for the in vivo study. Hesperidin and polyethylene glycol (PEG) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Because hesperidin is water-insoluble, it was dissolved in 70% PEG which is a widely used solvent for water-insoluble compounds in in vivo study [13]. Experimental protocol All experimental procedures conformed to the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996), and the Committee on Animal Research at Ajou Medical Center, Ajou University (Suwon, Republic of Korea), approved the study. Male Sprague-Dawley rats (weight, 250300 g) were anaesthetized with an intraperitoneal injection of ketamine (100 mg/kg) and xylazine (10 mg/kg) before surgery. The body temperature of the rats was maintained at 3760.5uC during surgery by using a thermostatically controlled warming plate as described previously [14]. In the loss-of-function study, ischaemiainduced myocardial injury was induced by ligating the LAD artery as described previously [9,14]. Sham-operated control group (sham) underwent the same surgical procedures except that the suture placed under the left anterior descending was not tied. At 4 weeks after LAD occlusion, rats were euthanized by CO2 inhalation for heart isolation. LV was used for staining experiments. Infarct and peri-infarct zone of left ventricle were used for gelatin zymography and wester (...truncated)