Abstract:AimTo explore the effect of arecoline (Are) on the formation of foam cells induced by the oxidized low density lipoprotein (ox-LDL) and analyze the possible mechanisms.MethodsRAW 264.7 macrophages were incubated by ox-LDL (50 mg/L) for 48 h to induce foam cells.Arecoline (10-6mol/L, 10-5mol/L and 10-4mol/L) was added for 48 h at the same time.The cellular lipid accumulation was examined by oi1 red staining.The cellular contents of total cholesterol (TC) and free cholesterol (FC) were detected by high performance liquid chromatography assays.Cholesterol efflux from macrophages was examined by ³H labed cholesterol.The expressions of ATP-binding cassette transporter A1 (ABCA1) were detected by Real-time PCR and Western blot.ResultsOil red-staining positive cells were found and macrophages were filled with lipid droplet in foam cells model group.Compared with the foam cells, arecoline (10-5 and 10-4 mol/L) significantly decreased the amount of oil red-staining positive cells and the contents of lipid droplet of foam cells.Compared with thecontrol macrophages group, the contents of TC, FC, cholesteryl ester (CE) and CE/TC ratio in model group were significantly increased in foam cells model group.Arecoline (10-5 and 10-4mol/L)decreased the contents of TC, FC, cholesteryl ester (CE) and CE/TC ratio, and increased significantly cholesterol efflux and expression of ABCA1 compared with foam cells model group.ConclusionArecoline inhibits the formation of foam cell induced by ox-LDL and up-regulates of expression ABCA1 in RAW 264.7 macrophage-derived foam cells.

Abstract:AimTo observe the effects of soluble epoxide hydrolase inhibitors t-AUCB on the uptake and degradation of lipid in mouse macrophage.MethodsRAW264.7 mouse macrophage was cultured, then t-AUCB in various concentration(1, 10, 50 and 100 μmol/L) were added for 24 hours, or incubated with peroxisome proliferators activated receptor gamma (PPARγ) antagonist GW9662 (5 μmol/L).0 μmol/L t-AUCB treated group was taken as empty control.After then, the uptake and degradation of oxidized low density lipoprotein (ox-LDL) in cells were detected by radioligand assay.The mRNA and protein expression of CD36 were determined by real-time PCR and Western blot .Resultst-AUCB could dose-dependently increase the uptake and degradation of ox-LDL in mouse macrophage.After stimulated with 0, 1, 10, 50 and 100 μmol/L t-AUCB, the uptake level of ox-LDL were 414.96±46.71 μg/g, 519.54±47.7 μg/g, 629.04±37.97 μg/g,720.66± 48.58 μg/g, 881.57±68.44 μg/g, and the degradation of ox-LDL were 16180.23±967.28 μg/g, 17369.62±478.34 μg/g, 21794.85±689.36 μg/g, 27883.03±712.25 μg/g, 30194.61±635.71 μg/g.However, after incubation of GW9662, the uptake and degradation of ox-LDL with 100 μmol/L t-AUCB

Abstract:AimTo study the effect of 2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucoside (TSG) on expression of adhesion molecules P-selectin, E-selectin induced by H₂O₂ in human umbilical vein endothelial cells.MethodsThe expression of P-selectin, E-selectin mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) respectively.Results Treating endothelial cells with 200 μmol/L H₂O₂, the expression of E-selectin, P-selectin mRNA and protein level were significantly higher (p<0.01).Pretreated with TSG for 4 h, the expression of P-selectin, E-selectin mRNA and protein induced by H₂O₂ were significantly inhibited in endothelial cells (p<0.05).ConclusionsTSG probably can protect human umbilical vein endothelial cells from oxidative stress by decreasing expression of adhesion molecules.

Abstract:AimTo explore whether edaravone(EDA), a novel free radical scavenger, protect H9c2 cells against chemical hypoxia-induced injury.MethodsH9c2 cells were treated with Cobalt chloride (CoCl₂) to set up a chemical hypoxia-induced cellular injury model.Cell viability was detected by cell counter kit (CCK-8).Changes in morphology and amount of apoptotic cells were observed by Hoechst 33258 staining; Intracellular level of reactive oxygen species (ROS) was measured by DCFH-DA staining and photofluorography; Mitochondrial membrane potential (MMP) was tested by JC-1 staining and photofluorography.ResultsExposure of H9c2 cells to 100～1000 μmol/L CoCl₂ for 24 h dose-dependently reduced cell viability.At the range from 12 to 36 h, 800 μmol/L CoCl₂ time-dependently inhibited cell viability.Pretreatment with 10 to 40 μmol/L EDA or with NAC (a ROS scavenger) at 500 to 2000 μmol/L for 1 h prior to exposure to 800 μmol/L CoCl₂ for 24 h dose-dependently blocked the inhibition of cell viability by CoCl₂.Preconditioning with 40 μmol/L EDA for 1 h prior to exposure of H9c2 cells to 800 μmol/L CoCl₂ inhibited not only CoCl₂-induced overproduction of ROS, but also the apoptotic effect and MMP loss induced by CoCl₂.ConclusionsEDA can protect H9c2 cells against CoCl₂-induced injury, which may be associated with its antioxidant effect and protection of MMP.

Abstract:AimTo investigate the possible signaling pathway in angiopoietin-1 (Ang-1) regulated inflammatory factors of endothelial progenitor cells.MethodsWe transformated Ang-1 into tumor necrosis factor-α (TNF-α) induced endothelial progenitor cell (EPC), used the specific inhibitor pyrrolidine dithio carbamate (PDTC) to inhibit nuclear factor-kappa B (NF-κB).Western blot was used to detect the protein expression of Ang-1 and NF-κB, real-time polymerase chain reaction (real-time PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression of intercellular cell adhesion molecule-1 (ICAM-1) and vascular cellular adhesion molecule-1 (VCAM-1) in EPC.ResultsAng-1 protein expressed while no significant NF-κB protein expressed in EPC.Real-time PCR and ELISA show that compared to the TNF-α group, the expressions of ICAM-1 and VCAM-1 in Ang-1 group, PDTC group and Ang-1＋PDTC group significantly decreased (p<0.05), but there were no significant differences among the three groups (p＞0.05).ConclusionsAng-1 maybe affects adhesion molecules expression in TNF-α induced EPC through NF-κB signaling pathway.

Abstract:AimTo screen the genes that were related to atherosclerosis and ageing through the gene chip, and verified by qPCR further.MethodsExpression profiling of thoracic aortas of rats at 3 to 28 months of age was analysed by array.Then these genes in rats were detected by realtime PCR, and the carotid artery morphology change was observed by microscopy.ResultsWe got 155 genes (157 gene probes) that differentially expressed during the rat aging, and 39 genes which responded to caloric restriction (CR) and high calories (HC).These genes involved in glycometabolism, lipid metabolism, cell adhesion, oxidation, etc.qPCR result showed that CR can induce significant expression of several genes, including Ldha, Gyg, Hk2, Slc2a4, Dlat, Pex19, Cyld, Psmb4, and down-regulate Vcam1, Icam1, F2r, Tln1, Cyb5r3, Hspd1, Acvrl1 and Bgn expression,while the HC played opposite effect, which were consistent with the results of cDNA microarray.The carotid artery became markedly thickened after HC diet.Conclusion There is a group of genes that play a cross-talking network during vascular ageing and atherosclerosis, which expression responds to CR or HC and may be a consequence of atherosclerosis.

Abstract:AimTo investigate the effects of adiponectin on calcification in cultured cardiomyocytes via reducing endoplasmic reticulum stress-mediated apoptosis in vitro.MethodsPrimary cultured cardiomyocytes were obtained from neonatal rats by collagenase II digestion method.The α-actin expression as molecular marker of the cardiomyocytes was observed by immunocytochemistry.The cells cultured for 72 h were used in the experiment and divided into groups randomly: control group, calcified myocardial cells (CMC) group, adiponectin＋CMC (3, 10, 20, 30 mg/L) groups.The morphological changes of the cardiomyocytes were observed under phase contracted microscope.CMC was prepared by incubation with β-glycerophosphate (BGP).Calcification was confirmed by Von Kossa staining and Alizarin Red S.Calcium content and alkaline phosphatases (ALP).Alkaline phosphatase was measured by spectrophotometric measurement of p-nitorphenol release.Osteocalcin was detected by radioimmunoassay.The content of lactate dehydrogenase (LDH) was measured.The cardiomyocyte apoptosis was detected by flow cytometry.The expression levels of glucose regulated protein 78 (GRP78)/cysteine-containing aspartate-specific proteases-12 (Caspase-12)/osteoprotegerin (OPG) mRNA were detected by quantitative real-time polymerase chain reaction (qRT-PCR).ResultsThe immunocytochemical stain for smooth muscle α-actin confirmed cardiomyocytes phenotype and the multicellular nodules spontaneously

Abstract:AimTo explore the effect of tobacco smoke on hydrogen sulfide (H₂S)/cystathionine-γ-lyase (CSE) system in rat thoracic aorta.Methods10-week-old SD rats for the study were randomly divided into control group, short-term smoking group, mid-term smoking group and long-term smoking group.Passive cigarette smoking rat model was established by the method of tobacco smoked suction.The concentrations of H₂S in the serum was measured with sensitive sulfur electrode.The expression of CSE in the thoracic aortic smooth muscle was observed with the methods of immunohistochemical staining and quantitative microscopic image analysis.ResultsH₂S concentration between short-term smoking group and control group had no significant difference (p>0.05).H₂S concentration of long-term smoking group and mid-term smoking group were respectively lower than that of the control group and short-term smoking group (p<0.01).H₂S concentration of long-term smoking group was significantly lower than that of the mid-term smoking group (p<0.05).The concentration of H₂S reduced with the extension of time of tobacco smoke suction and was time-dependent.The CSE expression in the thoracic aortic between short-term smoking group and control group had no significant difference (p>0.05), expression in long-term smoking group and mid-term smoking group were respectively lower than in control group and short-term smoking group (p<0.05), and in long-term smoking group was significantly lower than in the mid-term smoking group (p<0.05).The CSE and SUR-2 expression reduced with the extension of time of tobacco smoked suction, and was time-dependent.ConclusionsTobacco smoke can significantly lower H₂S content in the rat serum and CSE expression in the rat thoracic aorta.

Abstract:AimTo observe the effect of glucagon-like peptide-1 (GLP-1) receptor agonists exenatide on nuclear factor-κB (NF-κB) activity as well as the expression of its downstream inflammatory cytokins such as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) induced by high glucose in human umbilical vein endothelial cells (HUVEC) and to explore mechanism of protective effect of exenatide on vascular complication of diabetes.MethodsHUVEC were cultured in vitro and incubated under serum-free conditions with 25 mmol/L glucose for 48 hours.Exenatide at different concentrations (10－8 mol/L, 10－7 mol/L and 10－6 mol/L) was added concurrently with glucose stimulation.Meanwhile ICAM-1 and VCAM-1 in the cell culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA).The expression of NF-κB p65, ICAM-1 and VCAM-1 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results25 mmol/L D-glucose significantly increased NF-κB p65 mRNA expression as well as the supernatant content and mRNA expression of ICAM-1 and VCAM-1 in HUVEC (p<0.01)， which was inhibited by different concentrations of exenatide (p<0.01)， in a dose-dependent manner.ConclusionsGLP-1 receptor agonist exenatide could improve endothelial dysfunction via inhibition of NF-κB activity and its downstream inflammatory response in vascular endothelial cells induced by high glucose in a dose-dependent manner, which could prevent and improve vascular complication of diabetes.

Abstract:AimTo discuss the role of endothelial dysfunction in the kidney damage induced by hyperuricemia.Methods36 male Wister rats were randomly divided into three groups, namely control group, hyperuricemia model group, benzbromarone treatment group(n=12).Application of Oteracil potassium(uricase inhibitor ) and high yeast feeding were used to establish the hyperuricemia model.In addition to the same treatment,the treatment group were added benzbromarone to control hypeuricemia.Control group were fed normal diet and tap water, at the weekend of 2,4,6, blood uric acid and creatinine level were tested.All rats were sacrificed at the end of 6 weeks, HE and Masson staining of renal organizations were applied to observe the pathological changes, and measure the interstitial fibrosis area.Endothelial nitric oxide synthase (eNOS), endothelin-1(ET-1) and hypoxia inducible factor-1α(HIF-1α) expressions in renal interstitial tubular were detected by immunohistochemistry.ResultsAfter 2 weeks the blood uric acid level in model group was higher than that of the control group and treatment group.At the end of 6 weeks, the serum creatinine level in the model group was much higher than that of the control group and the treatment group, there was no obvious difference between the two latter groups.The renal interstitial fibrosis was more serious than that of the normal control group and the treatment group.The nitric oxide synthase expression in the model group was significantly lower than that in the control group and treatment group, while both the endothelin-1 and hypoxia inducible factor-1α expression in the model group was significantly higher than that in the control group and treatment group.ConclusionHyperuricemia can reduce the sy-nthesis of nitric oxide synthase but increase the production of endothelin-1 in the renal interstitial vascular endothelial cells, thus affecting the endothelial function of renal interstitial vascular and leading to renal interstitial fibrosis through an mechanism of ischemia and hypoxia.

Abstract:AimTo explore the effects and relative mechanism of metformin on changes of apoptosis associated genes survivin, p27, c-myc, c-fos and pathology in the diabetic rat myocyte.MethodsThe diabetic rat model was established.35 diabetic rats were randomly allocated into metformin(MT) group and diabetes mellitus(DM) group.15 rats were selected as control group.Therapeutical effect of myocardial pathology was detected by optical microscope and pathologic chemical staining.Apoptosis associated genes survivin, p27, c-myc, c-fos mRNA and relative proteins in the myocardium were detected by RT-PCR and Western blot. ResultsPathology of the DM group: myocyte arranged irregularly, myocyte hypertrophy, distortion, enlarged intercellular space, increased extracellular matrix, enhanced inflammatory cell infiltration, myocyte edema and more interstitial fibroblasts were observed.The changes of the myocardium in the MT group were significantly relieved compared with those of the DM group, myocyte ranked in order, interstitial contents decreased, perivascular matrix decreased, fibroblast decreased, inflammatory cell infiltration reduced markedly.The pathology changes of DM group were related to changes of survivin, p27, c-fos mRNA and protein. c-fos mRNA and protein level in MT group had no significant difference compared with the control group (p>0.05), but it was significantly higher than that of DM group (p<0.05). Survivin, p27 and c-myc mRNA and protein level in MT group had no significant difference compared with the DM group (p>0.05).ConclusionsThe pathology changes of DM group is related to changes of survivin, p27, c-fos mRNA and protein. Metformin improves pathological changes of diabetic cardiomyopathy which may be related to change of c-fos mRNA and protein.Metformin’s protective effect may not be related to survivin, p27 and c-myc mRNA and protein.

Abstract:AimTo investigate the circulating CD4^＋CD25^＋CD127^lo regulatory T cells (Tregs) levels in patients with acute myocardial infarction (AMI) and controls.Furthermore, to determine the relation between initial Tregs levels and cardiac function, the severity of coronary atherosclerosis and prognosis.Methods114 patients with AMI (86 with ST-segment elevation myocardial infarction (STEMI) and 28 with non-ST-segment elevation myocardial infarction (NSTEMI)) and 38 normal subjects were enrolled.Initial circulating Tregs levels were measured using flow cytometry in all subjects at entry.In 114 patients with AMI, heart function was evaluated by serum B-type brain natriuretic peptide (BNP) and left ventricular ejection fraction (LVEF) determined by echocardiogram.SYNTAX score and the number of affected main vessels were obtained to reflect the severity of coronary atherosclerosis.At one-month after discharge, information about reoccurrence of angina pectoris and NYHA heart function classification was obtained by outpatient follow-up.ResultsLevels of circulating Tregs were lower in patients with AMI at admission in comparison with normal subjects (2.04%±1.20% vs 3.68%±1.15%, p＜0.01).No significant differences in circulating Tregs levels were observed

Abstract:AimTo study the relationship between serum paraoxonase-1(PON1) activity and glycosylated hemoglobin (HbA1c), coronary artery lesions in patients with coronary heart disease.MethodsA total of 133 patients with suspected diagnosis of coronary artery disease,according to the coronary angiography results, were divided into coronary heart disease group(n=101) and control group (n=32).According to HbA1c levels, 101 cases of coronary heart disease were divided into three sub-groups: group A 31 cases (HbA1c<6.5%), group B 41 cases (6.5%≤HbA1c<9%), group C 29 cases (HbA1c≥9%).Serum PON1 activity was measured spectrophotometrically, and coronary lesions were detected by Gensini integral result.PON1 activity and Gensini integral among groups were compared, relationship between PON1 activity and HbA1c level, coronary lesions degree was analysed.ResultsSerum PON1 activity in the coronary heart disease group (2.17+0.18) was significantly lower than that in control group (2.49+0.19,p<0.01); PON1 activity in group C declined obviously (p<0.01) and coronary stenosis Gensini integral increased (p<0.01) compared with group A, in group C PON1 activity had fallen (p<0.05) and coronary stenosis Gensini integral increased (p<0.05) compared with group B.The correlation analysis showed that the serum PON1 activity was negatively related to HbA1c levels (r=－0.534,p<0.01), and negatively related to Gensini integral (r=－0.742,p<0.01).

Abstract:AimTo Explore influence of anti-cardiolipin antibodies (ACA) and anti-neutrophil cytoplasmic autoantibodies (ANCA) on intima membrane thickness (IMT) and carotid artery stenosis of patients with acute cerebral infarction(ACI).Methods250 cases of the first onset ACI patients were continously collected, enzyme-linked immunosorbent assay was used to determine ACA in morning hollow serum of ACI patients, indirect immunofluorescence was used to determine ANCA.All selected patients were divided into three groups: the ACA positive group, ANCA positive group, negative control group (ACA and ANCA negative).Carotid artery was detect by ultrasonic, IMT and carotid artery stenosis degree were compared in three groups.ResultsLeft and right side IMT of ACA positive group and ANCA positive group were higher than negative control group in patients with ACI (p<0.001); Extracranial carotid artery stenosis rate in ACA positive group(88.68%) and ANCA positive group(85.71%) were higher than negative control group(54.71%,p<0.017).ConclusionACA, ANCA were related to IMT, carotid artery stenosis in ACI patients.

Abstract:AimTo evaluate the effect of fluvastatin sodium 40 mg BID on low density lipoprotein (LDLC) level and the tolerability in Chinese patients with stable angina or type 2 diabetes mellitus whose LDLC was failed to achieve LDLC goal according to Chinese lipid guideline(＞100 mg/dL) with fluvastatin sodium 40 mg QN.MethodsA multicenter, prospective and observational post marketing surveillance.251 dyslipidemia patients with stable angina or type 2 diabetes mellitus (T2DM) were enrolled in this study.These patients were failed to achieve their LDLC goal (＜100 mg/dL) by the treatment of fluvastatin sodium 40 mg QN for at least 4 weeks, and were up titrated to fluvastatin sodium 40 mg BID by the discretion of physicians.The follow up duration was 8 weeks.The primary efficacy variable was the percent change in LDLC from baseline at week 8.The Secondary variables were the proportion of patients reaching their targets in LDLC, the percent change from baseline in total cholesterol (TC), high density lipoprotein cholesterol (HDLC), triglyceride (TG )and the tolerability.ResultsTotally 251 intentions to treatment patients (ITT) were in this study.After 8 weeks fluvastatin 40 mg BID reduced LDLC by 27.3%, 25.3% and 28.9% in total patients, stable angina and T2DM respectively (p<0.001).59.4% patients reached their LDLC goal at the end of study, of those 52.9% patients with stable angina and 67.2% patients with T2DM reached their goal.TG was reduced by 17.02%, 16.50% and 17.78% in total patients, stable angina and T2DM respectively (p<0.001).4 (1.6%) patients experienced clinical significant aminotransferase (ALT/AST) elevated ＞3 times the upper limit of normal (ULN), among them 3 patients continued to use fluvastatin after reducing the dose to 40 mg QN. None of the cases were accompanied with any hepatic symptoms or bilirubin abnormal or albumin decrease.No myopathy events were reported and there were no elevations in creatine kinase levels ＞5× ULN.ConclusionsUp titration dose of fluvastatin from 40 mg/day to 80 mg/day allows good efficacy and safety, and provide an alternative treatment method for stable angina and T2DM patients whose LDLC failed to reach the goal by the treatment of 40 mg/day.

Abstract:AimTo assess the risk factors of cardiovascular disease （CVD）in the patients with rheumatoid arthritis (RA).MethodsWe analysed the prevalence of 179 patients with RA who met the revised American College of Rheumatology (ACR) criteria and evaluated the risk factors of CVD patients with RA.ResultsOne-factor analysis shows the risk in CVD of RA was associated with age, process of RA, intima-media thickness (IMT) , DAS28 disease activity score, number of involved extra-articular organs, rheumatoid factor(RF), platelet count, C-reactive protein (CRP) and total cholesterol （TC）level (p<0.05）.Multi-factor analysis indicates the risk in CVD of RA was associated with DAS28 disease activity score(OR=2.403), number of involved extra-articular organs(OR=1.197), RF(OR=2.510), platelet count(OR=1.166), CRP(OR=1.700) and TC (OR=1.351), and was not associated with other traditional CVD risk factors .ConclusionOur data suggest a higher risk of cardiovascular events patients with RA, providing the basis of treatment and prevention of CVD.

Abstract:AimTo study the changes of serum level of interleukin-1β(IL-1β), interleukin-1 receptor antagonist(IL-1Ra) and IL-1Ra/ IL-1β ratio in the patients with intracranial and extracranial atherosclerosis (As) and arterial stenosis(AS) as well as the relationship between IL-1β and IL-1Ra level.MethodsThe serum levels of IL-1β and IL-1Ra were detected in 30 patients of As, 30 AS and 30 healthy individuals by enzyme-linked immunosorbent assay and IL-1Ra/ IL-1β ratio were calculated.ResultsCompared with the control, the IL-1β level in patients with As and AS was significantly higher, but the IL-1Ra level and IL-1Ra/ IL-1β ratio was significantly lower.The levels of serum IL-1β,IL-1Ra in AS group had no difference compared with As group(p>0.05),but the IL-1Ra/IL-1β ratio was significantly lower(p<0.05).There was correlation between IL-1β and IL-1Ra levels in patients with AS and healthy, but there was no correlation between IL-1β and IL-1Ra levels in patients with As. ConclusionsThe IL-1β might accelerate the progress of As, but IL-1Ra might suppress it.The immune maladjustment of IL-1β and IL-1Ra might be correlative to the severity of As, and the IL-1Ra/ IL-1β ratio might be an available index for the severity of As.

Abstract:Diabetes which is due to absolutely lack of insulin(Type Ⅰ diabetes) or just relatively lack of insulin (Type Ⅱ diabetes) in vivo is a metabolic syndrome, and one of its main features is hyperglycemia.Now the optimized treatment of diabetes is insulin therapy, but the role of high levels of insulin on atherosclerosis remains controversial because diabetes were often complicated by atherosclerosis.Therefore, it is of importance in diabetic atherosclerosis therapy to illustrate the effect of high level of insulin on the cellular lipid balance in hyperglycemia condition.

Abstract:Dyslipidemia is a definite risk factor of cardiovascular disease (CVD).Despite the increased prevalence of dyslipidemia in the elderly, consideration about side effect and safety of lipid therapy persists in this group.We discuss characteristics of dyslipidemia, change of pharmacokinetics and pharmacodynamics in the elderly, and evaluate safety of statins on liver, muscle, kidney, nerve system and tumor in the elderly by the evidence of a series of clinical trials.

Abstract:Proprotein convertase subtilisin/kexin type 9 (PCSK9) is closely related to modulate plasma cholesterol metabolism as newly discovered, and belongs to the proprotein convertase (PC) family.PCSK9 is a protein that reduces risk of coronary artery disease, peripheral vascular disease, cerebrovascular disease, nervous system diseases and diabetes by lowering serum low density lipoprotein cholesterol (LDLC) levels.Recent studies have shown that PCSK9 polymorphisms may modulate plasma cholesterol levels either positively or negatively.Recognition of these polymorphisms may have clinical significance in assessing severity of disease, prognosis, or response to drug therapy.This review focuses on recent advances of the influence of PCSK9 polymorphisms on LDLC and clinical advance of PCSK9.