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GW28-e0330 Cardio-protective effects of Qishen Granule on sarcoplasmic reticulum Ca2+ handling in heart failure rats
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JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY, VOL. 70, NO. 16, SUPPL C, 2017
whether EGCG can also inhibit the apoptosis of vascular smooth muscle cells (VSMCs), as well as its molecular mechanisms remain unknown. The present study aimed to elucidate the role and mechanism of EGCG in protecting against H2O2-induced apoptosis in mouse VSMCs. METHODS VSMCs were separated from abdominal aorta of mice and the subculture cells at passages 3-8 were used in all the experiments. VSMCs were treated with 10m M, 50mM, 100mM, 150mM of EGCG for 2 hours prior to treatment with 200m M H2O2 of 30min. RESULTS Treatment with H2O2 significantly decreased the cell viability and induced apoptosis of VSMCs, which were attenuated by pretreatment with EGCG. In particular, EGCG pretreatment significantly inhibited the H2O2-induced upregulation of cleaved forms of caspase-3, caspase-8, and caspase-9, Bax, CathepsinD, and downregulation of Bcl-2. Moreover, the antioxidation effect of EGCG on VSMCs was determined to be associated with the 67kD laminin receptor (67LR). CONCLUSIONS In summary, our results demonstrated that EGCG improved cell viability and protected VSMCs against oxidative stress through both extrinsic and intrinsic pathways, while 67LR is likely to be an active and key receptor of EGCG. We thus provide a novel molecular mechanism of EGCG in inhibiting H2O2-induced apoptosis in VSMCs, as well as its function in preventing the development of atherosclerosis. GW28-e0328 Qishen Granule attenuates inflammation-induced myocardial apoptosis through inhibiting cyclooxygenase2 (COX2) pathway Linghui Lu,1 Yong Wang,2 Jing Wang,3,4 Qiyan Wang,2 Yi Zhang,3,4 Qian Zhang,2 Yan Wu,5 Chun Li,3 Wei Wang1 1 School of Traditional Chinese Medicine, Beijing University of Chinese Medicine; 2School of Life Sciences, Beijing University of Chinese Medicine; 3Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine; 4School of Chinese Materia Medica, Beijing University of Chinese Medicine; 5Center of Scientific Experiment, Beijing University of Chinese Medicine OBJECTIVES To investigate the effect of QSG on inflammationmediated myocardial apoptosis and to explore the underlying pharmacological mechanisms. METHODS In vitro apoptotic model was induced by culturing H9C2 cardiomyocytes with supernatant of lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage. Anti-apoptosis effect of QSG was evaluated by cell viability assay, Hoechest33258 and Rhodamine123 staining. In vivo HF mice were induced by left anterior descending (LAD) coronary artery ligation after pre-administration of QSG or Celecoxib for 7 days. Heart functions were assessed by echocardiography and apoptotic cardiomyocytes were detected by TdT mediated dUTP Nick End Labeling (TUNEL) staining 24h after acute myocardial infarction (AMI). The critical inflammation mediators and proteins in apoptotic pathway were quantified by western-blot. RESULTS In vitro study showed that QSG increased cell viability, inhibited cell apoptosis, reduced the expressions of cyclooxygenase2 (COX2) and p-nuclear factor kappa B (p-NFkB), and down-regulated ratio of Bax/B-cell lymphoma-2 (Bcl-2). In vivo study showed that QSG improved heart function, ameliorated myocardial inflammation and inhibited apoptosis. Expression of COX2 was inhibited by QSG. Factors in apoptotic signaling pathways, including Bcl-2, Bax, P53, Fas ligand (FasL) and Caspases, were regulated by QSG.
METHODS HF rat models were induced by left anterior descending coronary artery ligation surgery and high-fat diet feeding. Rats were divided into sham, model, QSG and positive control groups. The therapeutic effects of QSG were evaluated by echocardiography and blood lipid testing. Intracellular Ca2þ concentration and Sarco-endoplasmic reticulum ATPase 2a (SERCA2a) activity were detected by specific Assay Kits. Expressions of the critical mediators in SR Ca2þ handling were evaluated by western-blot and RT-qPCR. RESULTS HF model of rats developed ventricular remodeling accompanied with calcium overload and defective Ca 2þ release-uptake cycling in cardiomyocytes. Treatment with QSG improved contractive function, attenuated ventricular remodeling and reduced the basal intracellular Ca 2þ level. QSG prevented defective Ca2þ leak by attenuating hyperphosphorylation of Ryanodine Receptor 2, inhibiting expression of protein kinase A and up-regulated transcriptional expression of phosphatase 1. QSG also restored Ca2þ uptake by up-regulating expression and activity of SERCA2a and promoting phosphorylation of phospholamban. CONCLUSIONS QSG restored SR Ca2þ cycling in HF rats and served as an ideal alternative drug for treating HF. GW28-e0353 Effect of Renal Denervation on Sympathetic Nerve System and Renin–Angiotensin–Aldosterone System after Acute Myocardial Infarction in Chinese Mini-swine Bin Zhang,1 Yongjian Wu1 1 Fuwai Hospital, Chinese Academy of Medical Science, Beijing, China OBJECTIVES To evaluate the effect of RND on the systemic, renal and cardiac sympathetic nerve system (SNS) and Renin–Angiotensin– Aldosterone System (RAAS) after an acute myocardial infarction in Chinese Mini-swine. METHODS Anterior MI was induced in 24 Chinese mini-swine by transient balloon occlusion of left anterior descending coronary artery for 1.5 hours. After reperfusion for 2 hours, the swine were randomly assigned to receive bilateral catheter-based RDN (the RDN group) or sham-denervation (the sham group) and were studied up to 8 weeks. The activity of the systemic, renal and cardiac sympathetic nerve would be determined by measuring the concentration of noradrenaline (NE), epinephrine (E) in peripheral vein plasma, venae renales plasma and tissue at baseline，directly after RDN and after 8 weeks. In parallel, the activity of RAAS would be determined by measuring plasma and tissue concentration of renin (PRA), angiotensin-II (AII) and aldosterone (Ald). RESULTS 7 swines died in the modeling process, 3 in RDN group and 4 in sham group. There were no differences between two groups in the peripheral vein plasma and venae renales plasma concentration of NE, E, PRA, AII, Ald at baseline. After 8 weeks, concentration of peripheral vein plasma Ald differed in the RDN and sham groups (0.080.03 vs. 0.19 0.5, p¼0.001). Venae renales PRA (2.05 1.73 vs.0.19 0.21, p¼0.037), E (0.070.01 vs.0.050.01, p¼0.048) significantly decreased compared with baseline in the RDN group. Both Venae renales PRA and AII raised post MI and fell to baseline post RDN. Furthermore, there were no differences between the RDN and sham group in PRA, AII, Adl of the cardiac tissue, nor in NE, E, PRA, AII, Adl in renal tissue.
CONCLUSIONS QSG treatment attenuates inflammation-induced apoptosis in a multi-components and multi-targets mode. QSG might be an ideal alternative drug for inhibiting myocardial apoptosis in HF.
CONCLUSIONS Venae renales plasma biochemical indicators are more sensitive compared with the peripheral vein in co-related with RDN process. Besides, more easily performed clinical measurements are needed to confirm the procedural success, i.e. that RDN indeed leads to sufficient disruption of renal nerves.
GW28-e0330 Cardio-protective effects of Qishen Granule on sarcoplasmic reticulum Ca2þ handling in heart failure rats
GW28-e0396 Abnormal Cx43 contributes to myocardial fibrosis in offspring rats exposed to Lipopolysaccharide during pregnancy
Linghui Lu,1 Chun Li,2 Qiyan Wang,3 Qian Zhang,3 Yi Zhang,2,4 Hui Meng,2,4 Yong Wang,3 Wei Wang1 1 School of Traditional Chinese Medicine, Beijing University of Chinese Medicine; 2Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine; 3School of Life Sciences, Beijing University of Chinese Medicine; 4School of Chinese Materia Medica, Beijing University of Chinese Medicine
Yan Wen,1 Ya Liu1 1 Institute of Materia Medica and Department of Pharmaceutics, College of Pharmacy, Third Military Medical University, Chongqing, PR China
OBJECTIVES To assess the effect of Qishen Granule (QSG) on sarcoplasmic reticulum (SR) Ca 2þ handling in heart failure (HF) model of rats and to explore the underlying molecular mechanisms.
OBJECTIVES Myocardial fibrosis (MF) is the common pathology of many cardiovascular diseases at the end stage, however, the underling mechanism remains unclear. Given Prenatal exposure to Lipopolysaccharide (LPS) produces hypertension in adult offspring rats, the present study was to explore the effects of prenatal inflammation on MF in the offspring rats and to further assess its susceptibility to cardiovascular diseases.
JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY, VOL. 70, NO. 16, SUPPL C, 2017
whether EGCG can also inhibit the apoptosis of vascular smooth muscle cells (VSMCs), as well as its molecular mechanisms remain unknown. The present study aimed to elucidate the role and mechanism of EGCG in protecting against H2O2-induced apoptosis in mouse VSMCs. METHODS VSMCs were separated from abdominal aorta of mice and the subculture cells at passages 3-8 were used in all the experiments. VSMCs were treated with 10m M, 50mM, 100mM, 150mM of EGCG for 2 hours prior to treatment with 200m M H2O2 of 30min. RESULTS Treatment with H2O2 significantly decreased the cell viability and induced apoptosis of VSMCs, which were attenuated by pretreatment with EGCG. In particular, EGCG pretreatment significantly inhibited the H2O2-induced upregulation of cleaved forms of caspase-3, caspase-8, and caspase-9, Bax, CathepsinD, and downregulation of Bcl-2. Moreover, the antioxidation effect of EGCG on VSMCs was determined to be associated with the 67kD laminin receptor (67LR). CONCLUSIONS In summary, our results demonstrated that EGCG improved cell viability and protected VSMCs against oxidative stress through both extrinsic and intrinsic pathways, while 67LR is likely to be an active and key receptor of EGCG. We thus provide a novel molecular mechanism of EGCG in inhibiting H2O2-induced apoptosis in VSMCs, as well as its function in preventing the development of atherosclerosis. GW28-e0328 Qishen Granule attenuates inflammation-induced myocardial apoptosis through inhibiting cyclooxygenase2 (COX2) pathway Linghui Lu,1 Yong Wang,2 Jing Wang,3,4 Qiyan Wang,2 Yi Zhang,3,4 Qian Zhang,2 Yan Wu,5 Chun Li,3 Wei Wang1 1 School of Traditional Chinese Medicine, Beijing University of Chinese Medicine; 2School of Life Sciences, Beijing University of Chinese Medicine; 3Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine; 4School of Chinese Materia Medica, Beijing University of Chinese Medicine; 5Center of Scientific Experiment, Beijing University of Chinese Medicine OBJECTIVES To investigate the effect of QSG on inflammationmediated myocardial apoptosis and to explore the underlying pharmacological mechanisms. METHODS In vitro apoptotic model was induced by culturing H9C2 cardiomyocytes with supernatant of lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage. Anti-apoptosis effect of QSG was evaluated by cell viability assay, Hoechest33258 and Rhodamine123 staining. In vivo HF mice were induced by left anterior descending (LAD) coronary artery ligation after pre-administration of QSG or Celecoxib for 7 days. Heart functions were assessed by echocardiography and apoptotic cardiomyocytes were detected by TdT mediated dUTP Nick End Labeling (TUNEL) staining 24h after acute myocardial infarction (AMI). The critical inflammation mediators and proteins in apoptotic pathway were quantified by western-blot. RESULTS In vitro study showed that QSG increased cell viability, inhibited cell apoptosis, reduced the expressions of cyclooxygenase2 (COX2) and p-nuclear factor kappa B (p-NFkB), and down-regulated ratio of Bax/B-cell lymphoma-2 (Bcl-2). In vivo study showed that QSG improved heart function, ameliorated myocardial inflammation and inhibited apoptosis. Expression of COX2 was inhibited by QSG. Factors in apoptotic signaling pathways, including Bcl-2, Bax, P53, Fas ligand (FasL) and Caspases, were regulated by QSG.
METHODS HF rat models were induced by left anterior descending coronary artery ligation surgery and high-fat diet feeding. Rats were divided into sham, model, QSG and positive control groups. The therapeutic effects of QSG were evaluated by echocardiography and blood lipid testing. Intracellular Ca2þ concentration and Sarco-endoplasmic reticulum ATPase 2a (SERCA2a) activity were detected by specific Assay Kits. Expressions of the critical mediators in SR Ca2þ handling were evaluated by western-blot and RT-qPCR. RESULTS HF model of rats developed ventricular remodeling accompanied with calcium overload and defective Ca 2þ release-uptake cycling in cardiomyocytes. Treatment with QSG improved contractive function, attenuated ventricular remodeling and reduced the basal intracellular Ca 2þ level. QSG prevented defective Ca2þ leak by attenuating hyperphosphorylation of Ryanodine Receptor 2, inhibiting expression of protein kinase A and up-regulated transcriptional expression of phosphatase 1. QSG also restored Ca2þ uptake by up-regulating expression and activity of SERCA2a and promoting phosphorylation of phospholamban. CONCLUSIONS QSG restored SR Ca2þ cycling in HF rats and served as an ideal alternative drug for treating HF. GW28-e0353 Effect of Renal Denervation on Sympathetic Nerve System and Renin–Angiotensin–Aldosterone System after Acute Myocardial Infarction in Chinese Mini-swine Bin Zhang,1 Yongjian Wu1 1 Fuwai Hospital, Chinese Academy of Medical Science, Beijing, China OBJECTIVES To evaluate the effect of RND on the systemic, renal and cardiac sympathetic nerve system (SNS) and Renin–Angiotensin– Aldosterone System (RAAS) after an acute myocardial infarction in Chinese Mini-swine. METHODS Anterior MI was induced in 24 Chinese mini-swine by transient balloon occlusion of left anterior descending coronary artery for 1.5 hours. After reperfusion for 2 hours, the swine were randomly assigned to receive bilateral catheter-based RDN (the RDN group) or sham-denervation (the sham group) and were studied up to 8 weeks. The activity of the systemic, renal and cardiac sympathetic nerve would be determined by measuring the concentration of noradrenaline (NE), epinephrine (E) in peripheral vein plasma, venae renales plasma and tissue at baseline，directly after RDN and after 8 weeks. In parallel, the activity of RAAS would be determined by measuring plasma and tissue concentration of renin (PRA), angiotensin-II (AII) and aldosterone (Ald). RESULTS 7 swines died in the modeling process, 3 in RDN group and 4 in sham group. There were no differences between two groups in the peripheral vein plasma and venae renales plasma concentration of NE, E, PRA, AII, Ald at baseline. After 8 weeks, concentration of peripheral vein plasma Ald differed in the RDN and sham groups (0.080.03 vs. 0.19 0.5, p¼0.001). Venae renales PRA (2.05 1.73 vs.0.19 0.21, p¼0.037), E (0.070.01 vs.0.050.01, p¼0.048) significantly decreased compared with baseline in the RDN group. Both Venae renales PRA and AII raised post MI and fell to baseline post RDN. Furthermore, there were no differences between the RDN and sham group in PRA, AII, Adl of the cardiac tissue, nor in NE, E, PRA, AII, Adl in renal tissue.
CONCLUSIONS QSG treatment attenuates inflammation-induced apoptosis in a multi-components and multi-targets mode. QSG might be an ideal alternative drug for inhibiting myocardial apoptosis in HF.
CONCLUSIONS Venae renales plasma biochemical indicators are more sensitive compared with the peripheral vein in co-related with RDN process. Besides, more easily performed clinical measurements are needed to confirm the procedural success, i.e. that RDN indeed leads to sufficient disruption of renal nerves.
GW28-e0330 Cardio-protective effects of Qishen Granule on sarcoplasmic reticulum Ca2þ handling in heart failure rats
GW28-e0396 Abnormal Cx43 contributes to myocardial fibrosis in offspring rats exposed to Lipopolysaccharide during pregnancy
Linghui Lu,1 Chun Li,2 Qiyan Wang,3 Qian Zhang,3 Yi Zhang,2,4 Hui Meng,2,4 Yong Wang,3 Wei Wang1 1 School of Traditional Chinese Medicine, Beijing University of Chinese Medicine; 2Modern Research Center for Traditional Chinese Medicine, Beijing University of Chinese Medicine; 3School of Life Sciences, Beijing University of Chinese Medicine; 4School of Chinese Materia Medica, Beijing University of Chinese Medicine
Yan Wen,1 Ya Liu1 1 Institute of Materia Medica and Department of Pharmaceutics, College of Pharmacy, Third Military Medical University, Chongqing, PR China
OBJECTIVES To assess the effect of Qishen Granule (QSG) on sarcoplasmic reticulum (SR) Ca 2þ handling in heart failure (HF) model of rats and to explore the underlying molecular mechanisms.
OBJECTIVES Myocardial fibrosis (MF) is the common pathology of many cardiovascular diseases at the end stage, however, the underling mechanism remains unclear. Given Prenatal exposure to Lipopolysaccharide (LPS) produces hypertension in adult offspring rats, the present study was to explore the effects of prenatal inflammation on MF in the offspring rats and to further assess its susceptibility to cardiovascular diseases.