CVIA Journal

[GW30-e0060]

Authors: Chao Li^1,2, Shijun Zhang³, Zhibo Gai^1,2, Yunlun Li¹

¹Shandong University of Traditional Chinese Medicine

²Department of Clinical Pharmacology and Toxicology, University Hospital of Zurich

³Shandong University of Traditional Chinese Medicine, Jinan, China

OBJECTIVES Obesity is a major contributor to myocardial cell apoptosis, fibrosis and ventricle hypertrophy, and associated with the increased risk of hypertrophic cardiomyopathy. Obeticholic acid (OCA), a farnesoid X receptor agonist, is a key regulator of lipid metabolism, inflammatory, fibrosis and metabolic pathways. This study was performed to investigate the effect and mechanism of OCA on obesity-induced myocardial injury.

METHODS C57Bl/6 mice were fed with a 45% high fat diet (HFD) or a standard diet. Biochemical parameters and myocardial pathological changes were examined. Energy metabolism in isolated working heart using radioactive was also tested to reveal the mechanism of myocardial injury. In vitro, 3D cell culture, mitochondria damage and ATP production of C2C12 cells cultured with palmitic acid (PA) in the absence or presence of OCA were tested.

RESULTS The body weight of HFD C57Bl/6 mice has increased by 22.7% compared with mice fed with normal diet. In addition, HFD-induced obese mice developed cardiac hypertrophy, fibrosis, inflammation, apoptosis, oxidative injury, which was rescued by OCA treatment. There was also a remarkably mitochondria damage in the obese mice and OCA prevented against the mitochondria damage. In vitro, 3D cell culture showed that PA reduced the myocardial contraction and viability. PA also increased the apoptosis rate of C2C12 cells and induced mitochondrial damage identified by the Tom20 level and ATP production assay. OCA showed a protective effect against PA-induced mitochondrial damage and myocardial damage. The results of energy metabolism in isolated working heart further indicated that the contribution of glucose oxidation to ATP production is lower than palmitate oxidation in HFD-induced obese mice heart, and OCA promoted glucose oxidation to protect against PA-induced mitochondrial damage.

CONCLUSIONS The present data suggested that OCA reduced the myocardial cell apoptosis, fibrosis and inflammation in the HFD-induced obese C57Bl/6 mice. OCA also protected cardiomyocytes against PA-induced mitochondria damage through promoting glucose oxidation. Our findings provide evidence for the protective role of OCA in myocardial cells.

[GW30-e0056]

Authors: Kun Lian, Ling Tao

Department of Cardiology, Xijing Hospital, Fourth Military Medical University

OBJECTIVES Diabetic patients are more sensitive tomyocardial ischemia-reperfusion (MI/R) injury. Branched-chain amino acids (BCAA) catabolism is defective and mitochondrial phosphatase 2C (PP2Cm) expression is reduced in the diabetic state. However, the role of PP2Cm and BCAA in diabetes with MI/R injury remains unclear. This study aims to determine the mechanism of reduced PP2Cm expression and investigate whether PP2Cm and BCAA have a cardioprotective effect in diabetes with MI/R injury.

METHODS C57BL/6 mice were fed a high-fat diet (HFD) and injected intraperitoneally with a dose of streptozotocin (25 mg/kg) twice to generate T2DM model mice. C57BL/6 mice (WT), type 2 diabetes (T2DM), PP2Cm–/– mice, were used in this study. For T2DM mice, PP2Cm-specific adenovirus was delivered to generate diabetic mice with PP2Cm overexpression. The T2DM mice were also treated with with BDK inhibitor BT2, while the PP2Cm–/– mice were treated with MnTBAP (manganese (III) tetrakis (4-benzoic acid)porphyrin chloride). Myocardial infarction/reperfusion (MI/R) was produced simultaneously in all types of mice. Additionally, WT and PP2Cm–/– mice treated with BT2 and expressing adenovirus were analyzed. H9C2 cells were treated with BCAA and BCKA. After H9C2 cells were treated with BCKA and underwent simulated ischemia-reperfusion (SI/R), BT2 and MnTBAP were added. Cardiac function, apoptosis, BCAA metabolism and oxidative damage were assayed.

RESULTS PP2Cm protein levels were significantly decreased in the diabetic heart. Under PP2Cm-overexpressing T2DM mice injury, cardiac function was improved due to a decrease of myocardial infarct size and the increase of LVEF. The Apoptosis rate was decreased as evidenced by Caspase-3 activity and the number of TUNEL positive cardiomyocytes. Cardiac BCAA and BCKA levels, as well as the ratio of p-BCKDE1α/BCKDE1α significantly increased (P<0.01) and BCKD activity significantly decreased (P<0.01) in T2DM mice. After BT2 treatment in T2DM mice with MI/R injury, the BCAA cataboliam defect was alleviated. At the same time, an improvement of Cardiac function and reduction of apoptosis was observed. In PP2Cm–/– mice, aBCAA catabolism defect and MI/R injury was observed. After PP2Cm overexpression and BT2 treatment, BCAA catabolism defect and MI/R injury was obviously alleviated. In PP2Cm–/– mice, oxidative damage was observed evident as an the increases in superoxide concentration, a decrease of MnSOD, complex I and III activities, and ATP levels as well as mitochondrial damage. Supplementation with MnTBAP obviously ameliorated this oxidative damage and MI/R injury in PP2Cm–/– mice. Treatment with BCKA (1.5-3 mM) resulted in significant decreases cell viability, and significant increases in the percentage of LDH release, apoptosis in H9C2 cells with SI/R injury. After BT2 and MnTBAP treatment, these effects were obviously alleviated.

CONCLUSIONS T2DM induced a downregulation of PP2Cm and reduced the defect of BCAA metabolism. PP2Cm directly mediated the defect of BCAA catabolism and oxidative damage. Overexpression PP2Cm alleviated MI/R injury by reducing the catabolism of BCAA and oxidative damage.

[GW30-e0053]

Authors: Jia Su, Xiaomin Chen

Department of Cardiology, Ningbo No. 1 Hospital, Ningbo, Zhejiang, P.R. China

OBJECTIVES The failure of therapeutic response to clopidogrel in platelet inhibition, which is called clopidogrel resistance (CR), is more likely to cause cardiovascular events. We aimed to study the contribution of promoter DNA methylation of paraoxonase 1 (PON1) to the risk of clopidogrel poor response.

METHODS Through VerifyNow P2Y12 assay, patient’ platelet functions were measured. Among 57 non-CR and 49 CR patients, the levels of DNA -methylation in four CpG dinucleotides on the PON1 promoter were tested using bisulfite pyrosequencing technology. Besides, the relative expression of PON1 mRNA was analysed by quantitative real-time PCR. Logistic regression was applied to investigate the interatcion of PON1 methylation and clinical -factors in CR.

RESULTS In the subgroup with dyslipidaemia, we discovered that higher CpG4 levels of the PON1 promoter indicated a poorer clopidogrel response (cases versus controls (%): 51.500±14.742 versus 43.308±10.891, =0.036), and the PON1 mRNA expression was reduced in CR patients. Additionally, the logistic regression indicated that higher level of albumin and the index of ALT were related with a lower risk of CR, and the index of AST as well as the quantity of stent may be positively associated with CR.

CONCLUSIONS The DNA methylation of CpG4 in the PON1 promoter would lead to a low expression of PON1 mRNA, which might induce clopidogrel resistance in the patients with dyslipidaemia, and the number of stents might be a risk for CR.

[GW30-e0052]

Authors: Jia Su, Xiaomin Chen

Department of Cardiology, Ningbo No. 1 Hospital, Ningbo, Zhejiang, P.R. China

OBJECTIVES MicroRNA (miRNA) can be used as predictive biomarkers for cardiovascular diseases, especially for acute myocardial infarction (AMI). However, few reports have focused on the value of exosomal miRNAs in the mechanism of the pathophysiological process from stable coronary artery disease (SCAD) to AMI.

METHODS Exosomes were isolated via ExoQuick precipitation after serum samples were collected. The exosomes were then identified by transmission electron microscopy (TEM), Western blotting, and nanoparticle-tracking analysis (NTA). The differential expression of miRNAs in exosomes from 6 AMI and 6 matching SCAD patients was screened using Agilent Human miRNA Microarrays. Target genes of the candidate miRNAs were predicted via an online miRNA database, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Further validation was conducted through quantitative real-time PCR (qRT-PCR) with 60 exosome (30 AMI and 30 SCAD) samples.

RESULTS The expression of 13 miRNAs (miRNA-4507, miRNA-3656, miRNA-6803-5p, miRNA-7108-5p, miRNA-6850-5p, miRNA-4486, miRNA-6741-5p, miRNA-1227-5p, miRNA-3195, miRNA-4634, miRNA-7975, miRNA-6798-3p, and miRNA-1915-3p) was significantly down-regulated in the AMI samples compared with the SCAD samples. In addition, we identified various target genes that are mainly involved in the pathways of cardiac rehabilitation and remodelling, such as the signalling pathways activated downstream by Nerve Growth Factor (NGF) and Fibroblast growth Factor Receptors (FGFRs). Validation of the expression of candidate miRNAs indicated that exosomal miRNA-1915-3p, miRNA-4507, and miRNA-3656 were significantly less expressed in AMI samples than in SCAD samples, and ROC curve (AUC) analysis showed that the expression of these miRNAs resulted in good predictive accuracy [miRNA-1915-3p (AUC 0.772); miRNA-4507 (AUC: 0.684); and miRNA-3656 (AUC: 0.771)], suggesting that serum exosomal miRNA-1915-3p, miRNA-4507, and miRNA-3656 might be predictive for AMI at an early stage. Correlation analysis revealed that the expression of miRNA-1915-3p was negatively correlated with the PLT (r=–0.479, P=0.038), the expression of miRNA-4507 was negatively correlated with LDL-c (r=–0.5, P=0.029), and the expression of miRNA-3656 was related to the LVEF (r=0.471, P=0.042).

CONCLUSIONS Exosomal miRNA-1915-3p, miRNA-4507, and miRNA-3656 might play an important role in the pathophysiology of acute myocardial infarction and could serve as clinical diagnostic biomarkers.

[GW30-e0051]

Authors: Wu Junyan¹, Wang Lei², Zhou Jiansheng³, Zhang Yanling¹

¹Institute of Cardiovascular Diseases, Taian Maternal and Children’s Hospital, Taian, China

²Department of Pediatric Surgery, Soochow University Affiliated Children’s Hospital, Suzhou, China

³John Curtin School of Medical Research, Australian National University, Australia

OBJECTIVES Hydronephrosis is commonly caused by an obstruction of the urine flow from the kidney. The condition involves a dilation and distention of the renal pelvis. Hydronephrosis may be either acute or chronic in nature. It was reported a postnatal infant had hypoplastic left heart with hydronephrosis, indicating that there may be a relationship between hydronephrosis and heart disease. However, whether hydronephrosis causes cardiac damage and affects the RAS in the heart remains unknown. Thus, we assessed BP, heart weight and the expression of components of the RAS in the heart in hydronephrotic mice treated with AT1 receptor blockade and ACE inhibitor.

METHODS Hydronephrosis was induced by left ureteral ligation in Balb/C mice except sham-operated animals. Blood pressure was measured by the tail-cuff method using photoelectric volume oscillometry. At postmortem, heart weight was balanced. The levels of cardiac ACE, ACE2 and Mas receptor were measured by RT-PCR and Western blot after treatment of losartan or enalapril. Plasma renin activity (PRA), Ang I and Ang II were measured by radioimmunoassay using commercial kits.

RESULTS In the normal kidney the tubules were intact while they disappeared in the hydronephrotic kidney. Blood pressure did not significantly change after the left ureteral ligation. Hydronephrosis led to an increase of ACE level and a decreased of ACE2 and Mas receptor in the heart. Losartan decreased cardiac ACE level, but ACE2 and Mas receptor levels significantly increased in hydronephrotic mice (P<0.01). Enalapril increased ACE2 levels (P<0.01), but did not affect Mas receptor in the heart. PRA decreased in hydronephrotic mice, but significantly increased by losartan or enalapril. Plasma Ang II level decreased in hydronephrotic mice (P<0.05). Administration of losartan was accompanied by a rise in plasma Ang I and Ang II concentrations in hydronephrotic animals (P<0.05). Enalapril also increased levels of Ang I (P<0.01) and Ang II (P<0.05) in the circulation.

CONCLUSIONS In this study, we found that Hydronephrosis increased cardiac ACE, suppressed ACE2 and Mas receptor levels. Furthermore, AT1 blockade caused sustained activation of cardiac ACE2 and Mas receptor, but ACE inhibitor had the limitation of such activation of Mas receptor in hydronephrotic animals. These findings may lead to an exciting new area in the clinical administration of AT1 receptor blockade. These results also suggest that activation of cardiac ACE2 by both enalapril and losartan may protect against the adverse effects of activated RAS and renal impairment. Thus, we propose that the change of cardiac ACE2 and Mas receptor expression induced by hydronephrosis can be an important target of strategies for preventing cardiovascular damage. The observations of the different molecular mechanisms of losartan and enalapril could be helpful for better options in the treatment of cardiovascular diseases.

This study was supported by Science and Technology Department of Shandong Province (No. 2016GSF201207) and the Natural Science Foundation of China (No. 81270336). *equal contribution; #Correspondence.

[GW30-e0038]
Authors: Yajuan Ni, Deng Jie, Liu Xin, Li Qing, Ni Yajuan

Department of Cardiology, The Second Affiliated Hospital of Xi’an Jiaotong University

OBJECTIVES Apoptosis of myocardial cells has been shown to be a critical step provoking heart failure. Studies indicated that mitochondrial oxidative stress and excessive production of mitochondrial ROS in heart failure was tightly linked to activation of apoptosis. ECH was used as a traditional Chinese herbal medicine, it has been shown to possess powerful ability of anti-oxidant and anti-apoptosis properties in kinds of cells, but whether it affects myocardial apoptosis in the heart failure remain unknown. The present study investigated the effects of ECH on heart failure and myocardial apoptosis of rats induced by ISO in vivo and on mitochondrial oxidative stress of AC-16 cells induced by ISO and explored the underlying mechanisms in vitro.

METHODS Heart failure rats were induced by ISO, ECH was treated by intraperitoneal injection, echocardiography was performed to evaluate heart function, TUNEL was used to detect myocardial apoptosis in vivo. In vitro AC-16 cells were cultured, mitochondrial oxidative stress was induced by ISO, ECH was pre-treated. Apoptotic cells were detected by flow cytometry, the level of mitochondrial ROS were measured by luminol chemiluminescence, 8-OHdG was used to evaluate the oxidative damage of mitochondrial DNA, mitochondrial membrane potential was detected by JC-1, carbonylation of mitochondrial proteins were measured using Elisa, mitochondrial lipid peroxidation were measured using TBARS assay, intracellular ROS were measured with flow cytometry.

RESULTS The results demonstrated that ECH significantly improved the heart function and reduced myocardial apoptosis of heart failure rats induced by ISO in vivo, and inhibited oxidative damage of mitochondrial DNA, protected mitochondrial membrane potential, reduced intracellular ROS, prevented carbonylation of mitochondrial proteins and mitochondrial lipid peroxidation, suppressed production of mitochondrial ROS, and -subsequently inhibited intracellular ROS of AC-16 cells induced by ISO in vitro.

CONCLUSIONS We concluded that ECH significantly inhibited myocardial apoptosis and improved heart function of heart failure rats induced by ISO via suppressing mitochondrial oxidative stress. It was suggested that ECH was a potential drug treatment for heart failure.

[GW30-e0030]
Authors: Xin Su, Shuai Wang, Daoquan Peng

The Second Xiangya Hospital of Central South University

OBJECTIVES Obesity is associated with a series of metabolic syndromes. The hallmark of obesity is excessive lipid storage in adipose tissue. As is known, the adipose tissue has such abundant adipose-derived mesenchymal stem cells (AMSCs), which can differentiate into mature adipocytes by imbalance between energy intake and expenditure.

Apolipoprotein A1 (ApoA1) is the major protein component of HDL. In addition to anti-atherogenic function of ApoA1, recent works focused on ApoA1 in affecting the process of obesity and the lipid metabolism in mature adipocytes, demonstrating that APOA1 gene SNPs were related to obesity and reduced plasma ApoA1 level was associated with increased prevalence of obesity. However, the underlying mechanisms are ambiguous. The aim of this study was to examine the anti-obesity effect of ApoA1 and the potential mechanisms by which ApoA1 influencing human AMSCs adipogenesis.

METHODS We isolated AMSCs from the epigastric adipocyte tissue of the patients underwent abdominal surgery. The pre-adipocytes were treated with adipogenesis medium and ApoA1 protein. Then we harvested cells at 7^th, 14^th days after adipogenesis. The following tests were performed separately: (1) effects of ApoA1 on the morphological changes of intracellular lipid droplets were observed by Oil red O staining under microscope; (2) effects of ApoA1 on the intracellular TG content were observed by spectrophotometry; (3) effects of ApoA1 on modulating the expression levels of the adipogenesis-related markers, such as C/EBPα, C/EBPβ, FABP4 and FAS, were detected by PCR and Western Blot; (4) by lentiviral transfection technology, we silenced and over-expressed the SORT1 gene in AMSCs. The function of sortilin in AMSCs adipogenesis was investigated and the effects of ApoA1 on sortilin expression were detected by PCR and Western blot; (5) the effect of apoA1 on adipogenesis was further detected in AMSCs with SORT1-silenced or over-expressed.

RESULTS The main results were listed as follows: (1) ApoA1 could reduce the amount of lipid droplets and decrease the TG content synergistically in adipocytes during the adipogenesis; (2) ApoA1 could down-regulate the gene and protein expression level of C/EBPα, C/EBPβ, FABP4 and FAS during the adipogenesis; (3) Sortilin plays an important role in AMSCs adipogenesis. Silencing SORT1 gene could promote excessive adipogenesis of AMSCs, while over-expression of SORT1 gene inhibits the AMSCs adipogenesis; (4) ApoA1 could up-regulate the gene and protein level of sortilin during the adipogenesis; (5) the effect of ApoA1 on inhibiting adipogenesis was attenuated in AMSCs with silencing SORT1 gene; however, there was no significant changes of the effect of ApoA1 on inhibiting adipogenesis in AMSCs with overexpressed SORT1 gene.

CONCLUSIONS In conclusions, our results confirm that sortilin plays an important role in the adipogenesis of human AMSCs. Furthermore, the results indicate that ApoA1 acts as a negative regulator of adipogenesis differentiation in human AMSCs through the inhibition of adipogenesis differentiation-related factors and the promotion the intracellular gene and expression level of sortilin. The present data provide insight into the mechanisms of the inhibitory effects of ApoA1 and suggest that the inhibitory activity of ApoA1 indicates a potential pharmacological intervention specifically directed toward obesity.

[GW30-e0029]

Authors: Xin Su, Daoquan Peng

The Second Xiangya Hospital of Central South University

OBJECTIVES Obesity is associated with metabolic syndromes. The hallmark of obesity is excessive lipid storage in adipose tissue. As is known, the adipose tissue has such abundant adipose-derived mesenchymal stem cells (AMSCs), which can differentiate into mature adipocytes by imbalance between energy intake and expenditure.

Apolipoprotein A5 (ApoA5) is a novel apolipoprotein. Recently, evidence indicates that lower plasma level of ApoA5 was found in obese subjects and was inversely correlated with BMI. However, the underlying mechanisms are ambiguous. On the other hand, it is noteworthy that ApoA5 could also modulate TG storage in hepatocytes, indicating a crucial intracellular role of ApoA5 in TG metabolism. Since adipocytes provide the largest storage depot for TG, we hypothesized apoA5 might also target to adipocytes and regulate TG storage. The aims of this research were to explore the effect of ApoA5 in AMSCs adipogenesis and the underlying mechanisms.

METHODS We isolated AMSCs from the epigastric adipocyte tissue of the patients underwent abdominal surgery. The pre-adipocytes were treated with adipogenesis medium and human recombinant ApoA5 protein. Then we harvested cells at 7^th, 14^th, 21^st days after adipogenesis. The following tests were performed: (1) effects of ApoA5 on morphological changes of intracellular lipid droplets were observed under microscope; (2) effects of ApoA5 on intracellular TG content were observed by spectrophotometry; (3) effects of ApoA1 on modulating the gene expression levels of the adipogenesis-related markers, such as C/EBPα, C/EBPβ, PPARγ, aP2 and FAS, were detected by PCR; (4) effects of ApoA1 on modulating the gene expression levels of CIDEC were detected by PCR; (5) distribution of ApoA5 and CIDEC were observed by confocal microscope; (6) ApoA5 antibody and CIDEC antibody were used for CO-IP observe whether ApoA5 interacts with CIDEC; (7) we silenced and over-expressed the CIDEC gene in AMSCs. The function of CIDEC in AMSCs adipogenesis was investigated and the effects of ApoA5 on CIDEC expression were detected; (8) the effect of ApoA5 on adipogenesis was further detected in AMSCs with CIDEC-silenced or over-expressed.

RESULTS The main results were listed as follows: (1) ApoA5 could reduce the amount of lipid droplets and decrease the TG content in adipocytes during the adipogenesis; (2) ApoA5 could down-regulate the gene expression level of C/EBPα, C/EBPβ, PPARγ, aP2 and FAS during the adipogenesis; (3) ApoA5 could down-regulate the gene and protein expression level of CIDEC during the adipogenesis; (4) ApoA5 co-localized and had interaction with CIDEC in the surface of lipid droplets; (5) the effect of ApoA5 on inhibiting adipogenesis was attenuated in AMSCs with SORT1 gene over-expression.

CONCLUSIONS In conclusions, our results confirm that CIDEC plays an important role in the adipogenesis of human AMSCs. Furthermore, the results indicate that apoA5 acts as a negative regulator of adipogenesis differentiation in human AMSCs through the inhibition of adipogenesis differentiation-related factors and the promotion the intracellular gene and expression level of CIDEC. The present data provide insight into the mechanisms of the inhibitory effects of ApoA5 and suggest that the inhibitory activity of ApoA5 indicates a potential pharmacological intervention specifically directed toward obesity.

[GW30-e0011]

Authors: Yarmolenko Olga, Bumeister Valentyna, Prykhodko Olga, Demikhova Nadiia, Bumeister Lina, Khotyeev Yegor, Sumy State University

OBJECTIVES Diabetes mellitus is one of the most common non-communicable diseases. Today, more than 382 million people with diabetes live on the planet. The dynamics of development is increasing every year. Diabetes mellitus is one of the main causes of cardiovascular pathologies. The purpose of our work was to establish the age-old peculiarities of the effect of alloxan diabetes on the heart of rats. The features of changes in internal organs are studied on experimental animal models in order to improve the methods of correction of adverse effects of diabetes mellitus on the body and, in particular, on the heart. One of the inductors of diabetes is alloxan.

METHODS The study was performed on 12 mature and 12 young white male rats, divided into two groups: control and experimental (6 in each). Animal retention and experiments were carried out in accordance with the requirements of the “General Ethical Principles of Animal Experiments”, adopted by the First National Congress on Bioethics (Kyiv, 2001). Animals of the experimental group administered once daily alloxan intraperitoneally at a dose of 40 mg/kg. From the 7^th to the 10^th day after the administration of alloxan, the level of glucose in the blood that was steadily elevated was measured. One month after the induction, the animals were withdrawn from the experiment with an anesthetized lung of decapitation. Hearts were digested according to the method of Avtandilov, separately weighed parts of the heart behind Muller. For histological examination, the ventricles were fixed in a 10% neutral formalin solution over the course of the day, dehydrated in alcohols of increasing concentration, and poured into paraffin. The sections of the myocardium were stained with hematoxylin-eosin and studied using a light microscope Olympus BH-2.

RESULTS Under conditions of alloxan diabetes in mature rats there is an increase in the heart mass by 41.09% (P<0.0001), left ventricle is 56% (P><0.0001), right ventricle is 31.46% (P><0.0001), the left ventricle area was 31.77% (P><0.0001), right ventricle 45.89% (P=0.0008). Atrial weight decreases by 64% (P><0.0001), the ventricular index is 14.94% (P=0.0014), the planimetric index is unreliable. Histologically, the polymorphism of nuclei of cardio-myocytes, local disorientation of muscle fibers and their cytolysis were revealed. Spacing gaps are expanded (stromal edema). Vessels of uneven filling: in some fields, the vessels are empty, in others – the aggregation of erythrocytes in vessels, capillary hyperemia, edema around the vessels. In young rats, an increase in left ventricular mass is observed at 33.48% (P=0.0327), left ventricular area is 18.22% (P=0.0061), right ventricle is 20% (P=0.0287). Atrial weight decreases by 38.78% (P=0.0035), ventricular index is 15.19% (P=0.0036). Other indicators are unreliable. Histologically – the polymorphism of the nuclei of cardiomyocytes, the disorientation of muscle fibers. Unidirectional contents of the vessel: in some fields the vessels are empty, in others the aggregation of red blood cells.

CONCLUSIONS In alloxan diabetes, an increase in the mass of the heart with an overwhelming left ventricular hypertrophy, cardiomyocytes and vascular disorders in the myocardium of experimental animals was detected. The violations of the structural components of the wall of the heart are detected. This indicates a low functional activity of the heart, which leads to cardiovascular pathologies.

[GW30-e0006]

Authors: Qing Wang, Huiming Zhang, Congping Su, Hui Luo, Wenchao Jiao, Shuzhen Guo

School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China

OBJECTIVES The aim of this study was to evaluate the protective effect of sweroside on excessive isoproterenol induced heart failure in mice and to explore its target and mechanism.

METHODS Heart failure mice model was established by subcutaneous injection of excessive isoprenaline. Mice were randomly divided into four groups including control group, model group, sweroside group and captopril group. Cardiac function was evaluated by echocardiography, myocardial pathological changes were detected by Hematoxylin and eosin staining (HE staining). Drug targets were predicted by BAT-MAN-TCM database, and protein expression was detected by Western blot.

RESULTS Compared with the control group, the left ventricle ejection fraction (LVEF) and left ventricle fractional shortening (LVFS) of the model group decreased by 37.84 and 48.04% respectively. Left ventricular internal systolic diameter (LVIDs) increased to about 3.06±0.51 mm (1.86±0.59 mm in the control group), with an increase of more than 50%. Compared with the model group, EF of sweroside group increased to 71.83±4.82%, which was close to the control group, FS increased by 71.79%, LVIDs decreased by 30.82%. The above data had statistical significance (P<0.01). In Histopathological examination, thinned left ventricular wall, thinned myocardial fiber, scattered necrosis of myocardial cells, as well as aggregation of inflammatory cells were found in model mice. The morphology of myocardial tissue were obtained approximate normal by sweroside. The BAT-MAN-TCM database suggests that sweroside may play a therapeutic role in heart failure through ATPase Na+/K+Transporting Subunit Alpha1(ATP1A1). Western blot results showed that ATP1A1 protein expression was down-regulated in the model group, and the content of ATP1A1 protein was increased in the sweroside group.

CONCLUSIONS Sweroside can significantly improve the heart failure induced by excessive isoproterenol in mice, and the mechanism may be related to the increase of ATP1A1 protein expression and the improvement of myocardial energy metabolism.