- Email: [email protected]
GW28-e0704 PCSK1 MUTANT MICE DISPLAY INCREASED APOA1 LEVEL AND DECREASED PLTP ACTIVITIY IN SERUM
C64
JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY, VOL. 70, NO. 16, SUPPL C, 2017
OBJECTIVES To study the preventive effect of Quercetin on silent mating type information regulation 2 homolog 1(SIRT1) and adenosine monophosphate-activated protein kinase (AMPK) in the rat with diabetics. METHODS Male Wistar rats were divided into 4 group, normal control (n＝10),diabetic rats with high-fat diet (n＝10), diabetic rats with highfat diet plus Quercetin (25 mg/kg$d, n＝10), diabetic rats with high-fat diet plus Quercetin (50 mg/kg$d, n＝10). the following indices of rats were measured respectively, Levels of blood creatine kinase (CK) and serum lactate dehydrogenase (LDH) as well as myocardial nonesterified fatty acids (NEFA) and collagen were determined using ultraviolet spectrophotometric, the concentration of myocardial fatty acid transport proteins (FATPs) and fatty acidb-oxidase (FA-b-oxidase) were measured by ELISA method, the protein expression of NF-kB, silent mating type information regulation 2 homolog 1 (SIRT1) and adenosine mon ophosphate-activated protein kinase (AMPK) were detected by westernblot. RESULTS Levels of CK and LDH as well as NEFA were remarkably decreased after treatment of TSG. Quercetin caused a significant increase in concentration of myocardial FATPs and FA-b-oxidase in DM rat model. In diabetic group, Cardiac tissue SOD and CAT activities were significantly lower than control group (p＜0.05). Quercetin caused significant increase in the SOD and CAT activities of DMþ Quercetin groups cardiac tissue compared to DM group (p＜0.05). In diabetic group, Cardiac tissue NF-kB and MDA levels were increased compared to control group (p＜0.05), and groups of DMþ Quercetin had lower NF-kB and MDA levels than diabetic group (p＜0.05).Quercetin dramatically restored the decrease of SIRT1, AMPKaand pAMPKa protein expression in diabetic rats. CONCLUSIONS As a conclusion, based on the results we obtained from this study, we determined in diabetic rats with high-fat diet, increased glucose levels and cardiac damage markers decreased significantly, after administration of Quercetin, that oxidative stress and NF-kB levels increased while SIRT1 levels decreased in the diabetic group. These findings indicate that the protective mechanisms of Quercetin against diabetic rats are involved in the alleviation of Inflammatory mediator’s injury and energy metabolism.
GW28-e0636 Evaluation of exercise-induced fatigue model in rats with HR using treadmill exercise with progressively increasing load Lijuan Hou,1 Xiwen Su,2 Yijun Bian,3 Wei Ji,1 Yanhang Zhang,1 Xiaoli Liu,1 Decai Qiao1 1 College of Physical Education and Sports, Beijing Normal University; 2 College of Life Sciences, Beijing Normal University; 3School of Computer Science and Technology, University of Science and Technology of China OBJECTIVES Exercise fatigue is a common phenomenon in athletics and a key factor that restricts an athlete’s performance level. The term “exercise fatigue” refers to a state in which the physiological processes of the body systems cannot continue at a certain level or the organism cannot maintain a predetermined exercise intensity. In exercise physiology, heart rate is an important index used to evaluate exercise load and ethological changes and can accurately reflect the immediate response of physical function to changes in exercise load. There is an important correlation between heart rate and ethological performance. Therefore, in this study, we used changes of ethological indexes and heart rate to classify the exercise stages and set up a fitting formula of exercise fatigue based on the rats’ treadmill exercise data. METHODS The rat exercise fatigue model was established using treadmill exercise with three progressively increasing loads (light to heavy). The rats were subjected to 5-day adaptive treadmill training after being fed for 3 days in the laboratory. A modified Bedford treadmill exercise with a progressively increasing load was used for the exercise fatigue model. There were three total load levels: level I, 8.2 m/min, lasting for 15 min; level II, 15 m/min, lasting for 15 min; level III: 20 m/min, lasting until exhaustion. Changes in ethological indexes and heart rate were observed and recorded by video and heart rate telemetry and statistical methods were used to analyze and test the data. RESULTS As expected, during the exercise fatigue process, the relative velocity of the rats decreased; their relative positions
gradually shifted backwards; the proportions of sound and electrical stimulation time as well as slippage time gradually increased; and the heart rate increased twice and was then maintained within a certain range. According to the above data, we divided the process into four stages. We also established a fitting formula as follows: Y ¼ 15.2548 þ 0.4346∙xa - 0.1154∙xb þ 0.6826∙xc þ 0.0044∙xa∙xb 0.0021∙xb∙xc - 0.0013∙xc∙xa - 0.0023∙xa2 - 0.0016∙xb2 (r2 ¼ 0.906); (xa, xb, and xc represent the cumulative time of sound stimulation, electrical stimulation, and slippage time within 600 s (in seconds); y represents a fatigue score within the 600 s that indicates the degree of exercise fatigue). CONCLUSIONS We quantified the ethological and heart rate changes in rats during the exercise fatigue process, and determined the proportion of time in its process of the whole practice. In addition, based on the ethological changes and heart rate changes of rats, we divided the exercise fatigue phenomenon into the spontaneous exercise, transition, early fatigue, and later fatigue stages, which laid a foundation for the quantitative evaluation of the ethology in the rat model of exercise fatigue with a progressively increasing load. Meanwhile, the effective ethological indexes were screened and we successfully established a simple nonlinear regression formula to evaluate the fatigue degree of rats with ethological indexes using a mathematical modeling method of polynomial fitting. Hopefully the results shown above will provide an effective reference for the formulation of evaluation standard of exercise fatigue degree to a certain extent(31401018). GW28-e0641 Transcatheter Mitral Valve Implantation Using the Mithos valve–Pre-Clinical Animal Study Yang Liu,1 Jian Yang,1 Jiayou Tang,1 Ping Jin,1 Shiqiang Yu1 1 Department of Cardiovascular Surgery, Xijing hospital, Fourth military medical university OBJECTIVES This pre-clinical study was aimed to evaluate the safety and feasibility of transcatheter mitral valve implantation using a novel designed Mithos valve (NewMed Medical Co., Ltd., Shanghai, China) in a porcine model. METHODS The Mithos valve is a self-expanding bioprosthesis with cross-linked bovine pericardial tissue tricuspid leaflets mounted inside a nitinol self-expanding frame designed for transcatheter mitral valve implantation. By using porcine animal models, Mithos valve was implanted through a transapical approach. Technical feasibility, safety, durability and function of the Mithos valve were evaluated with autopsy of explanted heart, ventriculography, intracardiac echocardiography (ICE), transthoracic echocardiography (TTE), multisliced CT (MSCT), histological and electron microscopic examinations in acute study and six months following the procedure. RESULTS Animal study of 26 swine demonstrated procedural success of 100%. Macroscopic evaluation of the explanted hearts demonstrated stable and secured positioning of the transcatheter valve, with full endothelialization of the valve leaflets and fabric coatings. Multimodality image studies including ventriculography, ICE, TTE, and MSCT showed excellent function and alignment of the valves, without coronary artery obstruction, left ventricular outflow tract obstruction, transvalvular gradients or obvious paravalvular leak. There was 7 mild-to-moderate degree of paravalvular leaks and no significant mitral regurgitation. Cardioscopy and macroscopic evaluation demonstrated stable and secure positioning of the Mithos valve without evidence of injury to the ventricular or atrial walls. Histological and electron microscopic examinations showed no obvious macro- or micro-calcification up at the 6-month follow-up. CONCLUSIONS Transcatheter mitral valve implantation using with Mithos valve is technically safe and feasible in pre-clinical studies, resulting in a stable and well-functioning mitral bioprosthesis. With a better understanding of preclinical knowledge, patient selection criteria and first-in-human studies will be addressed. GW28-e0704 PCSK1 MUTANT MICE DISPLAY INCREASED APOA1 LEVEL AND DECREASED PLTP ACTIVITIY IN SERUM Seungbum Choi,1 Aleksandra Aljakna,2 Ron Korstanje2 1 Gachon Cardiovascular Research Institute, Incheon, Republic of Korea; 2 The Jackson Laboratory, Bar Harbor, ME, USA
JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY, VOL. 70, NO. 16, SUPPL C, 2017
C65
OBJECTIVES Several proprotein convertase subtilisin/kexin (PCSK) members are involved in high-density lipoprotein (HDL) metabolism. We investigated the potential role of PCSK1 in HDL metabolism.
pathway and restraining phosphorylation of ERK signaling pathway.
METHODS A mouse model (PCSK1N222D) containing an amino acid substitution (N->D) at position 222 in PCSK1 was used to test whether HDL metabolism is affected.
GW28-e0754 High Cl- promotes VCAM-1 expression via NKCC-1-mediated PI3K/ AKT/ GSK3b/ IkBa/ NFkB pathway in high Nacl-exposed endothelial cells
RESULTS HDL cholesterol concentration and apolipoprotein A1 (APOA1) serum levels were compared between wild-type (PCSK1WT) and PCSK1N222D. HDL cholesterol concentration was not affected, but the APOA1 level was increased in the non-HDL fraction of PCSK1 N222D, indicating that the unbound APOA1 is increased. We hypothesized that some of the secreted APOA1 remains in the lipid-free form due to inefficient APOA1-lipidation mediated by two enzymes: phospholipid transfer protein (PLTP) and lecithin-cholesterol acyltransferase (LCAT). PLTP activity was lower in PCSK1N222D serum when APOA1 and HDL cholesterol concentrations were induced by a diet supplemented with vitamin D, suggesting that lower PLTP activity in PCSK1N222D serum is a cause of lipid-free APOA1 induction. CONCLUSIONS Our data suggests a role for PCSK1 in HDL metabolism and an interaction with PLTP. GW28-e0705 C-Reactive Protein Causes ERK1/2 Activation and Aggravates Myocardial Ischemia Reperfusion Injury in Primary Culture of Cardiomyocytes Weina Pei,1 Fan Liu,1 Haijuan Hu,1 Bing Xiao,1 Wei Cui1 1 Department of Cardiology, Second Hospital of Hebei Medical University, Shijiazhuang, Hebei OBJECTIVES Recent studies have indicated that C-reactive protein (CRP) is a causal agent in atherogenesis rather than merely a marker of inflammatory processes. However, the effects of CRP on myocardial ischemia reperfusion injury, an urgent problem to be solved, and the mechanisms underlying have not been determined. Furthermore, whether statins have protective effects on CRP-mediated ischemia reperfusion injury is still unknown. METHODS We simulated ischemia reperfusion using oxygen-glucose deprivation/ reoxygenation (OGD/R) in neonatal Sprague-dawley rat cardiomyocyte, which was induced by 3 hours of hypoxia plus glucose and serum deprivation, followed by 1 hour of reperfusion. Cell viability was tested with MTS assay and cardiomyocyte damage was evaluated by Lactate dehydrogenase (LDH) leakage. Mitochondrial membrane potential was determined by tetramethylrhodamine ethyl ester (TMRE) and mitochondrial permeability transition pore (mPTP) opening was measured by calcein/AM, using confocal laser scanning microscopy. In addition, we studied the signaling pathway underlying CRP-mediated ischemia reperfusion injury via western blot analysis. RESULTS Compared with the simple OGD/R group, after intervention with 10 m g/ml of CRP, the cell viability decreased markedly (82.36 % 6.18% versus 64.84% 4.06%, P¼0.0007) and the LDH leakage had a significant rise [(145.316.06) U/L versus (208.219.23) U/L, P¼0.0122]. We also found that CRP directly activated mPTP opening and reduced mitochondrial membrane potential during ischemia reperfusion. Furthermore, pretreatment with 1 mM of atorvastatin (Ator) before CRP intervention protected cardiomyocytes from ischemia reperfusion injury. Interestingly, the mitochondrial K ATP channel opener diazoxide and the mPTP inhibitor cyclosporin A can offset the effects of CRP on cell viability, LDH leakage and the fluorescence intensity values of calcein/AM and TMRE. Moreover, the expression of phosphorylation of extracellular-signal-regulated kinase (ERK) 1/2 was significantly higher after pre-treatment with CRP compared with the value simply through OGD/R (170.4%3.00% versus 93.53%1.94%, P<0.0001). Western blot analysis also revealed that Akt expression was markedly activated after co-treatment with Ator and CRP, compared with the level after simply CRP pretreatment (184.2%6.96% versus 122.7%5.30%, P¼0.0003). The difference was also significant in the change of phosphorylation of ERK between these two groups. CONCLUSIONS Our results suggested that CRP directly aggravates myocardial ischemia reperfusion injury in myocardial cells and may be mainly mediated by inhibiting mitoK ATP Channels and promoting mPTP Opening, but Ator do the opposite. Ator can reduce CRPinduced ischemia reperfusion injury. Furthermore, one of the mechanisms in CRP-induced ischemia reperfusion injury may relate to the sustained activation of the ERK1/2 signaling pathway. Ator protected cardiomyocytes mainly through activating AKt signaling
Zhenjie Gu,1 Yongbiao Fang,1 Shaohua Wang,1 Yangxin Chen1 1 Department of Cardiology, Sun Yat-sen Memorial Hospital of Sun Yatsen University, Guangzhou, China OBJECTIVES High salt diet (HSD) contributed to the development of cardiovascular diseases (CVDs), however, with the mechanisms remained elusive. It was found that HSD promoted inflammatory responses including cardiovascular inflammation in vivo, and inflammation drove CVDs’ development. HSD-induced inflammation was closely associated with high Nacl resulted from HSD. HSD-induced cardiovascular inflammation might be regulated by pro-inflammatory endothelial cells (ECs). The expression of adhesion molecules including VCAM-1, which mediates leucocytes infiltration, underlies the pro-inflammatory ECs. VCAM-1 was important for the initiation of atherosclerosis, a common inflammatory CVD. The present study was aimed to determine whether high Nacl and HSD could induce VCAM-1 expression in vivo and in vitro, as well as its related mechanisms. METHODS Human ECs were exposed to high Nacl, and VCAM-1 expression was examined. Naþ and Cl- were substituted for NMDGþ and gluconate with mannitol as isotonic control in order to determine which part was critical for VCAM-1 induction. Then its associated signaling pathway was studied by means of specific inhibitors. In vivo, SD rats were fed with HSD or normal salt diet (NSD), and aorta intima was collected followed by investigation of VCAM-1 expression and the related signaling molecules activation. RESULTS High Nacl remarkably upregulated VCAM-1 expression in ECs, which was dependent on high Cl- but not high Naþ or osmolality. PI3K/ AKT/ GSK3b/ IkBa/ NFkB pathway activation mediated high Nacl-induced VCAM-1 expression. Meanwhile, HSD promoted VCAM-1 expression and the above mentioned signaling molecules activation in aorta intima of SD rats in comparison of NSD. Bumetanide as the specific inhibitor for Na-K-Cl cotransporter (NKCC-1) significantly suppressed high Nacl-induced PI3K/ AKT/ GSK3b/ IkBa/ NFkB pathway activation and VCAM-1 expression. CONCLUSIONS High Cl- activated PI3K/ AKT/ GSK3b/ IkBa/ NFkB pathway via NKCC-1, thereby promoting VCAM-1 expression in high Nacl-stimulated ECs. NKCC-1 might be a feasible target in HSDassociated CVDs intervention. GW28-e0862 Comparison of Immune Suppression Strategies for Transplantation of human Embryonic Stem Cell-Derived Cardiovascular Progenitor Cells in Postmyocardial Infarcted Nonhuman Primates Keyang Zhu,1,3 Qiang Wu,2,6 Cheng Ni,1,3 Peng Zhang,2,6 Zhiwei Zhong,1,3 Yan Wu,1,3 Yingchao Wang,1,3 Yinchuan Xu,1,3 Minjian Kong,4 Haifeng Cheng,4 Fengjiang Zhang,5 Qi Chen,5 Yi Li,5 Yun Jiang,2,6 Jianyi Zhang,7 Huangtian Yang,2 Xinyang Hu,1,3 Jianan Wang1,3 1 Department of Cardiology, Second Affiliated Hospital, College of Medicine, Zhejiang University; 2Key Laboratory of Stem Cell Biology and Laboratory of Molecular Cardiology, Institute of Health Sciences, Shanghai Jiao Tong University School of Medicine & Shanghai Institutes for Biological Sciences; 3Cardiovascular Key Laboratory of Zhejiang Province; 4Department of Cardiovascular Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University; 5 Department of Anesthesiology, Second Affiliated Hospital, College of Medicine, Zhejiang University; 6University of Chinese Academy of sciences; 7Department of Biomedical Engineering, University of Alabama OBJECTIVES Human pluripotent stem cell-derived cardiac lineage cells hold great promise for cardiac repair following injury. The evaluation of their therapeutic properties for both efficiency and safety in large animal studies are prerequisite for clinic applications. However, immune rejection is one of the major barriers in preclinical xenotransplantation animal study, especially in nonhuman primates. To determine suitable immunosuppression strategies for human pluripotent stem cell-derived cardiac lineage cells in nonhuman primates.
JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY, VOL. 70, NO. 16, SUPPL C, 2017
OBJECTIVES To study the preventive effect of Quercetin on silent mating type information regulation 2 homolog 1(SIRT1) and adenosine monophosphate-activated protein kinase (AMPK) in the rat with diabetics. METHODS Male Wistar rats were divided into 4 group, normal control (n＝10),diabetic rats with high-fat diet (n＝10), diabetic rats with highfat diet plus Quercetin (25 mg/kg$d, n＝10), diabetic rats with high-fat diet plus Quercetin (50 mg/kg$d, n＝10). the following indices of rats were measured respectively, Levels of blood creatine kinase (CK) and serum lactate dehydrogenase (LDH) as well as myocardial nonesterified fatty acids (NEFA) and collagen were determined using ultraviolet spectrophotometric, the concentration of myocardial fatty acid transport proteins (FATPs) and fatty acidb-oxidase (FA-b-oxidase) were measured by ELISA method, the protein expression of NF-kB, silent mating type information regulation 2 homolog 1 (SIRT1) and adenosine mon ophosphate-activated protein kinase (AMPK) were detected by westernblot. RESULTS Levels of CK and LDH as well as NEFA were remarkably decreased after treatment of TSG. Quercetin caused a significant increase in concentration of myocardial FATPs and FA-b-oxidase in DM rat model. In diabetic group, Cardiac tissue SOD and CAT activities were significantly lower than control group (p＜0.05). Quercetin caused significant increase in the SOD and CAT activities of DMþ Quercetin groups cardiac tissue compared to DM group (p＜0.05). In diabetic group, Cardiac tissue NF-kB and MDA levels were increased compared to control group (p＜0.05), and groups of DMþ Quercetin had lower NF-kB and MDA levels than diabetic group (p＜0.05).Quercetin dramatically restored the decrease of SIRT1, AMPKaand pAMPKa protein expression in diabetic rats. CONCLUSIONS As a conclusion, based on the results we obtained from this study, we determined in diabetic rats with high-fat diet, increased glucose levels and cardiac damage markers decreased significantly, after administration of Quercetin, that oxidative stress and NF-kB levels increased while SIRT1 levels decreased in the diabetic group. These findings indicate that the protective mechanisms of Quercetin against diabetic rats are involved in the alleviation of Inflammatory mediator’s injury and energy metabolism.
GW28-e0636 Evaluation of exercise-induced fatigue model in rats with HR using treadmill exercise with progressively increasing load Lijuan Hou,1 Xiwen Su,2 Yijun Bian,3 Wei Ji,1 Yanhang Zhang,1 Xiaoli Liu,1 Decai Qiao1 1 College of Physical Education and Sports, Beijing Normal University; 2 College of Life Sciences, Beijing Normal University; 3School of Computer Science and Technology, University of Science and Technology of China OBJECTIVES Exercise fatigue is a common phenomenon in athletics and a key factor that restricts an athlete’s performance level. The term “exercise fatigue” refers to a state in which the physiological processes of the body systems cannot continue at a certain level or the organism cannot maintain a predetermined exercise intensity. In exercise physiology, heart rate is an important index used to evaluate exercise load and ethological changes and can accurately reflect the immediate response of physical function to changes in exercise load. There is an important correlation between heart rate and ethological performance. Therefore, in this study, we used changes of ethological indexes and heart rate to classify the exercise stages and set up a fitting formula of exercise fatigue based on the rats’ treadmill exercise data. METHODS The rat exercise fatigue model was established using treadmill exercise with three progressively increasing loads (light to heavy). The rats were subjected to 5-day adaptive treadmill training after being fed for 3 days in the laboratory. A modified Bedford treadmill exercise with a progressively increasing load was used for the exercise fatigue model. There were three total load levels: level I, 8.2 m/min, lasting for 15 min; level II, 15 m/min, lasting for 15 min; level III: 20 m/min, lasting until exhaustion. Changes in ethological indexes and heart rate were observed and recorded by video and heart rate telemetry and statistical methods were used to analyze and test the data. RESULTS As expected, during the exercise fatigue process, the relative velocity of the rats decreased; their relative positions
gradually shifted backwards; the proportions of sound and electrical stimulation time as well as slippage time gradually increased; and the heart rate increased twice and was then maintained within a certain range. According to the above data, we divided the process into four stages. We also established a fitting formula as follows: Y ¼ 15.2548 þ 0.4346∙xa - 0.1154∙xb þ 0.6826∙xc þ 0.0044∙xa∙xb 0.0021∙xb∙xc - 0.0013∙xc∙xa - 0.0023∙xa2 - 0.0016∙xb2 (r2 ¼ 0.906); (xa, xb, and xc represent the cumulative time of sound stimulation, electrical stimulation, and slippage time within 600 s (in seconds); y represents a fatigue score within the 600 s that indicates the degree of exercise fatigue). CONCLUSIONS We quantified the ethological and heart rate changes in rats during the exercise fatigue process, and determined the proportion of time in its process of the whole practice. In addition, based on the ethological changes and heart rate changes of rats, we divided the exercise fatigue phenomenon into the spontaneous exercise, transition, early fatigue, and later fatigue stages, which laid a foundation for the quantitative evaluation of the ethology in the rat model of exercise fatigue with a progressively increasing load. Meanwhile, the effective ethological indexes were screened and we successfully established a simple nonlinear regression formula to evaluate the fatigue degree of rats with ethological indexes using a mathematical modeling method of polynomial fitting. Hopefully the results shown above will provide an effective reference for the formulation of evaluation standard of exercise fatigue degree to a certain extent(31401018). GW28-e0641 Transcatheter Mitral Valve Implantation Using the Mithos valve–Pre-Clinical Animal Study Yang Liu,1 Jian Yang,1 Jiayou Tang,1 Ping Jin,1 Shiqiang Yu1 1 Department of Cardiovascular Surgery, Xijing hospital, Fourth military medical university OBJECTIVES This pre-clinical study was aimed to evaluate the safety and feasibility of transcatheter mitral valve implantation using a novel designed Mithos valve (NewMed Medical Co., Ltd., Shanghai, China) in a porcine model. METHODS The Mithos valve is a self-expanding bioprosthesis with cross-linked bovine pericardial tissue tricuspid leaflets mounted inside a nitinol self-expanding frame designed for transcatheter mitral valve implantation. By using porcine animal models, Mithos valve was implanted through a transapical approach. Technical feasibility, safety, durability and function of the Mithos valve were evaluated with autopsy of explanted heart, ventriculography, intracardiac echocardiography (ICE), transthoracic echocardiography (TTE), multisliced CT (MSCT), histological and electron microscopic examinations in acute study and six months following the procedure. RESULTS Animal study of 26 swine demonstrated procedural success of 100%. Macroscopic evaluation of the explanted hearts demonstrated stable and secured positioning of the transcatheter valve, with full endothelialization of the valve leaflets and fabric coatings. Multimodality image studies including ventriculography, ICE, TTE, and MSCT showed excellent function and alignment of the valves, without coronary artery obstruction, left ventricular outflow tract obstruction, transvalvular gradients or obvious paravalvular leak. There was 7 mild-to-moderate degree of paravalvular leaks and no significant mitral regurgitation. Cardioscopy and macroscopic evaluation demonstrated stable and secure positioning of the Mithos valve without evidence of injury to the ventricular or atrial walls. Histological and electron microscopic examinations showed no obvious macro- or micro-calcification up at the 6-month follow-up. CONCLUSIONS Transcatheter mitral valve implantation using with Mithos valve is technically safe and feasible in pre-clinical studies, resulting in a stable and well-functioning mitral bioprosthesis. With a better understanding of preclinical knowledge, patient selection criteria and first-in-human studies will be addressed. GW28-e0704 PCSK1 MUTANT MICE DISPLAY INCREASED APOA1 LEVEL AND DECREASED PLTP ACTIVITIY IN SERUM Seungbum Choi,1 Aleksandra Aljakna,2 Ron Korstanje2 1 Gachon Cardiovascular Research Institute, Incheon, Republic of Korea; 2 The Jackson Laboratory, Bar Harbor, ME, USA
JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY, VOL. 70, NO. 16, SUPPL C, 2017
C65
OBJECTIVES Several proprotein convertase subtilisin/kexin (PCSK) members are involved in high-density lipoprotein (HDL) metabolism. We investigated the potential role of PCSK1 in HDL metabolism.
pathway and restraining phosphorylation of ERK signaling pathway.
METHODS A mouse model (PCSK1N222D) containing an amino acid substitution (N->D) at position 222 in PCSK1 was used to test whether HDL metabolism is affected.
GW28-e0754 High Cl- promotes VCAM-1 expression via NKCC-1-mediated PI3K/ AKT/ GSK3b/ IkBa/ NFkB pathway in high Nacl-exposed endothelial cells
RESULTS HDL cholesterol concentration and apolipoprotein A1 (APOA1) serum levels were compared between wild-type (PCSK1WT) and PCSK1N222D. HDL cholesterol concentration was not affected, but the APOA1 level was increased in the non-HDL fraction of PCSK1 N222D, indicating that the unbound APOA1 is increased. We hypothesized that some of the secreted APOA1 remains in the lipid-free form due to inefficient APOA1-lipidation mediated by two enzymes: phospholipid transfer protein (PLTP) and lecithin-cholesterol acyltransferase (LCAT). PLTP activity was lower in PCSK1N222D serum when APOA1 and HDL cholesterol concentrations were induced by a diet supplemented with vitamin D, suggesting that lower PLTP activity in PCSK1N222D serum is a cause of lipid-free APOA1 induction. CONCLUSIONS Our data suggests a role for PCSK1 in HDL metabolism and an interaction with PLTP. GW28-e0705 C-Reactive Protein Causes ERK1/2 Activation and Aggravates Myocardial Ischemia Reperfusion Injury in Primary Culture of Cardiomyocytes Weina Pei,1 Fan Liu,1 Haijuan Hu,1 Bing Xiao,1 Wei Cui1 1 Department of Cardiology, Second Hospital of Hebei Medical University, Shijiazhuang, Hebei OBJECTIVES Recent studies have indicated that C-reactive protein (CRP) is a causal agent in atherogenesis rather than merely a marker of inflammatory processes. However, the effects of CRP on myocardial ischemia reperfusion injury, an urgent problem to be solved, and the mechanisms underlying have not been determined. Furthermore, whether statins have protective effects on CRP-mediated ischemia reperfusion injury is still unknown. METHODS We simulated ischemia reperfusion using oxygen-glucose deprivation/ reoxygenation (OGD/R) in neonatal Sprague-dawley rat cardiomyocyte, which was induced by 3 hours of hypoxia plus glucose and serum deprivation, followed by 1 hour of reperfusion. Cell viability was tested with MTS assay and cardiomyocyte damage was evaluated by Lactate dehydrogenase (LDH) leakage. Mitochondrial membrane potential was determined by tetramethylrhodamine ethyl ester (TMRE) and mitochondrial permeability transition pore (mPTP) opening was measured by calcein/AM, using confocal laser scanning microscopy. In addition, we studied the signaling pathway underlying CRP-mediated ischemia reperfusion injury via western blot analysis. RESULTS Compared with the simple OGD/R group, after intervention with 10 m g/ml of CRP, the cell viability decreased markedly (82.36 % 6.18% versus 64.84% 4.06%, P¼0.0007) and the LDH leakage had a significant rise [(145.316.06) U/L versus (208.219.23) U/L, P¼0.0122]. We also found that CRP directly activated mPTP opening and reduced mitochondrial membrane potential during ischemia reperfusion. Furthermore, pretreatment with 1 mM of atorvastatin (Ator) before CRP intervention protected cardiomyocytes from ischemia reperfusion injury. Interestingly, the mitochondrial K ATP channel opener diazoxide and the mPTP inhibitor cyclosporin A can offset the effects of CRP on cell viability, LDH leakage and the fluorescence intensity values of calcein/AM and TMRE. Moreover, the expression of phosphorylation of extracellular-signal-regulated kinase (ERK) 1/2 was significantly higher after pre-treatment with CRP compared with the value simply through OGD/R (170.4%3.00% versus 93.53%1.94%, P<0.0001). Western blot analysis also revealed that Akt expression was markedly activated after co-treatment with Ator and CRP, compared with the level after simply CRP pretreatment (184.2%6.96% versus 122.7%5.30%, P¼0.0003). The difference was also significant in the change of phosphorylation of ERK between these two groups. CONCLUSIONS Our results suggested that CRP directly aggravates myocardial ischemia reperfusion injury in myocardial cells and may be mainly mediated by inhibiting mitoK ATP Channels and promoting mPTP Opening, but Ator do the opposite. Ator can reduce CRPinduced ischemia reperfusion injury. Furthermore, one of the mechanisms in CRP-induced ischemia reperfusion injury may relate to the sustained activation of the ERK1/2 signaling pathway. Ator protected cardiomyocytes mainly through activating AKt signaling
Zhenjie Gu,1 Yongbiao Fang,1 Shaohua Wang,1 Yangxin Chen1 1 Department of Cardiology, Sun Yat-sen Memorial Hospital of Sun Yatsen University, Guangzhou, China OBJECTIVES High salt diet (HSD) contributed to the development of cardiovascular diseases (CVDs), however, with the mechanisms remained elusive. It was found that HSD promoted inflammatory responses including cardiovascular inflammation in vivo, and inflammation drove CVDs’ development. HSD-induced inflammation was closely associated with high Nacl resulted from HSD. HSD-induced cardiovascular inflammation might be regulated by pro-inflammatory endothelial cells (ECs). The expression of adhesion molecules including VCAM-1, which mediates leucocytes infiltration, underlies the pro-inflammatory ECs. VCAM-1 was important for the initiation of atherosclerosis, a common inflammatory CVD. The present study was aimed to determine whether high Nacl and HSD could induce VCAM-1 expression in vivo and in vitro, as well as its related mechanisms. METHODS Human ECs were exposed to high Nacl, and VCAM-1 expression was examined. Naþ and Cl- were substituted for NMDGþ and gluconate with mannitol as isotonic control in order to determine which part was critical for VCAM-1 induction. Then its associated signaling pathway was studied by means of specific inhibitors. In vivo, SD rats were fed with HSD or normal salt diet (NSD), and aorta intima was collected followed by investigation of VCAM-1 expression and the related signaling molecules activation. RESULTS High Nacl remarkably upregulated VCAM-1 expression in ECs, which was dependent on high Cl- but not high Naþ or osmolality. PI3K/ AKT/ GSK3b/ IkBa/ NFkB pathway activation mediated high Nacl-induced VCAM-1 expression. Meanwhile, HSD promoted VCAM-1 expression and the above mentioned signaling molecules activation in aorta intima of SD rats in comparison of NSD. Bumetanide as the specific inhibitor for Na-K-Cl cotransporter (NKCC-1) significantly suppressed high Nacl-induced PI3K/ AKT/ GSK3b/ IkBa/ NFkB pathway activation and VCAM-1 expression. CONCLUSIONS High Cl- activated PI3K/ AKT/ GSK3b/ IkBa/ NFkB pathway via NKCC-1, thereby promoting VCAM-1 expression in high Nacl-stimulated ECs. NKCC-1 might be a feasible target in HSDassociated CVDs intervention. GW28-e0862 Comparison of Immune Suppression Strategies for Transplantation of human Embryonic Stem Cell-Derived Cardiovascular Progenitor Cells in Postmyocardial Infarcted Nonhuman Primates Keyang Zhu,1,3 Qiang Wu,2,6 Cheng Ni,1,3 Peng Zhang,2,6 Zhiwei Zhong,1,3 Yan Wu,1,3 Yingchao Wang,1,3 Yinchuan Xu,1,3 Minjian Kong,4 Haifeng Cheng,4 Fengjiang Zhang,5 Qi Chen,5 Yi Li,5 Yun Jiang,2,6 Jianyi Zhang,7 Huangtian Yang,2 Xinyang Hu,1,3 Jianan Wang1,3 1 Department of Cardiology, Second Affiliated Hospital, College of Medicine, Zhejiang University; 2Key Laboratory of Stem Cell Biology and Laboratory of Molecular Cardiology, Institute of Health Sciences, Shanghai Jiao Tong University School of Medicine & Shanghai Institutes for Biological Sciences; 3Cardiovascular Key Laboratory of Zhejiang Province; 4Department of Cardiovascular Surgery, Second Affiliated Hospital, College of Medicine, Zhejiang University; 5 Department of Anesthesiology, Second Affiliated Hospital, College of Medicine, Zhejiang University; 6University of Chinese Academy of sciences; 7Department of Biomedical Engineering, University of Alabama OBJECTIVES Human pluripotent stem cell-derived cardiac lineage cells hold great promise for cardiac repair following injury. The evaluation of their therapeutic properties for both efficiency and safety in large animal studies are prerequisite for clinic applications. However, immune rejection is one of the major barriers in preclinical xenotransplantation animal study, especially in nonhuman primates. To determine suitable immunosuppression strategies for human pluripotent stem cell-derived cardiac lineage cells in nonhuman primates.