Results == == 3

Results == == 3. 1 . ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) before and after the XG treatment and histopathologic changes were also examined. Myocardial cell apoptosis was detected by TUNEL staining. The expression of related genes and proteins were analyzed using immunohistochemical staining. == Results == Compared to those in the control group, rats in XG treated groups showed significantly improved cardiac function and milder cardiac histopathological changes, lower cardiomyocyte apoptosis index, higher expression of Bcl-2 and lower expression of Bax. == Conclusions == Administration of XG improves cardiac function and histopathological changes in rats with doxorubicin-induced cardiotoxicity. These effects are associated with inhibition of cardiomyocyte apoptosis, perhaps via regulation of Bcl-2 and Bax protein expression. Keywords: Apoptosis, Doxorubicin, Heart failure, Herbal medicine == 1 . Introduction == Xinfuli Granule (XG) is a compound traditional Chinese medicine consists of extracts from Radix astragali, Radix ginseng, Salvia miltiorrhiza, Scirpus fluviatilis, Rhizoma Alismatis, Angelica sinensis, Semen lepidii, Fructus chaenomelis, Semen arecae, and Ophiopogon japonicusformulation, which has been clinically used for the treatment of chronic heart failure (HF) for more than 50 years in Fuwai Hospital (Beijing, China). Our previous studies have demonstrated that XG could improve cardiac function in patients with heart failure, particularly in those patients with idiopathic dilated cardiomyopathy (DCM).[1]However , the underlying mechanisms of the treatment effects of XG is not clear. Apoptosis plays important role in the development and progress of HF. Both ischaemic and idiopathic DCM are associated with apoptosis.[2],[3]Recently, several studies have indicated that some traditional Chinese medicine (TCM) with similar formula as XG may attenuate ischemia/reperfusion injury and myocardial injury by suppressing apoptosis.[4],[5]Our present study used a rat model of doxorubicin induced cardiotoxicity to explore the cardioprotective effects of XG on cardiac function and the underlying mechanisms, focusing on the regulation of cell apoptosis. == 2 . Methods == == 2 . 1 . Animal model and grouping == Adult male JMS-17-2 Sprague-Dawley rats (weighing 200220 g) were provided by the Animal Center of Academy of Military Medical Sciences. The rats were kept in clean and quiet environment with room temperature 22 1 C, humidity 40 5%, and a 12 h light/dark cycle, and had free access to standard diet and water. They were treated with intraperitoneal injection of DOX saline solution (2. 5 mg/kg per week) for six weeks, then the live rats were randomly divided into four groups which received intragastrically administration of normal saline (control group) or low dosage of XG (0. 675 g/kg per day, XG-L group, n= 12), medium dosage of XG (1. 35 g/kg per day, XG-M group, n= 12) and high dosage of XG (2. 7 g/kg per day, XG-H group, n= 12) for six weeks. Twelve normal rats were also selected for normal group. All experimental procedures were approved by the ethic committee for animal use of Academy of Military Medical Sciences and were performed in accordance with their guidelines. == 2 . 2 . Echocardiography measurements == Before and after the XG treatment, Transthoracic echocardiography was performed with a 12-MHz phased-array transducer (Sonos 7500, Phillips). Two-dimensional parasternal long axis views were obtained in M-mode at the papillary muscle level. Left ventricular fractional shortening (LVFS) and left ventricular ejection fraction (LVEF) were calculated. The parameters were measured over three consecutive cardiac cycles. == 2 . 3. Tissue sample preparation == Rats were fasted for 12 Tmem1 h and then were sacrificed under deep anesthesia. The JMS-17-2 hearts were excised and immediately, rinsed with pre-cooling saline. The hearts were weighed to calculate the ratio of heart weight to body weight. After fixed 18 h in 10% maldehyde solution, the anterior wall of the hearts were embed in paraffin wax and cut into 4 m slices for subsequent apoptosis and immunohistochemical analysis. == 2 . 4. Histopathological examination == The Paraffin-embedded slices were stained with hematoxylin-eosin (HE) for morphologic examination. The extent of myocardial damage was evaluated in 200-fold magnification by light microscopy (Leica, Germany). == 2 . 5. Terminal deoxynucleotidyl transferase dUTP Nick end labeling (TUNEL) == The cell apoptosis rate was determined JMS-17-2 by TUNEL according to manufacturer’s instructions (Boster Biological Engineering Co. Ltd, Wuhan, China). Four micrographs were randomly selected. To assess the fraction of apoptotic.