Abstract:Aim To a review the prevention with probucol in subjects with high risk of cardiovascular disease. Methods The relevant information on the outcome events of probucol treatment was extracted from papers of Multifactorial Primary Prevention Trial of Vascular Disease in Finland to analyze the effect of probucol on coronary events. Results Probucol,especially when combined with clofibrate,markedly reduced high density lipoprotein cholesterol. The number of coronary events was higher than predictive values in those men who had been treated with clofibrate and beta-blockers but it is lower with probucol added. Five years cumulative incidence of coronary events is different in group with probucol added and without probucol added (1.0% vs 3.6%,P=0.040). Taking into account the impact of different drugs,there were significant association between probucol and coronary events (RR was 0.186,95%CI was 0.053 to 0.656,P=0.008). Conclusion Although probucol decreased high density lipoprotein cholesterol levels,if cholesterol levels remained high,other drugs combined with probucot can stiu reduce coronary events in high risk subjects.

Abstract:Aim To study the feasibility and mechanism of probucol in resolution of monosodium urate monohydrate (MSU) crystal-induced arthritis. Methods A rat model of acute gouty arthritis was induced with the injection of MSU into rat ankle joint cavity. The effects of probucol on arthritis were investigated through observing clinical features,the synovial fluid white blood cell count and interleukin-1β(IL-1β). IL-1β concentration of culture medium was observed after the 18 h intervention with probucol in THP-1 monocytes while stimulated by MSU. Results The degree of joint swelling decreased gradually from probucol low-dose group to high-dose group and there was signifanct difference between mid-dose and hign-dose probucol groups and model group at 72h(P<0.05). The synovial fluid white blood cell count and IL-1β declined in varying degrees in probucol groups compar with the model group. Synovial fluid white blood cell count of probucol mid-dose and high-dose group decreased significantly,comparing with the model group (P<0.01,P<0.001). IL-1β of probucol high-dose group decreased significantly comparing with the model group (P<0.05). Probucol group showed no inhibition on IL-1β secretion of the THP-1 human monocyte cells. Conclusion The mid-dose and hign-dose probucol can effectively control acute gouty arthritis attack and the effect is dependent on dose,but the effect is not produced by inhibiting IL-1β secretion of THP-1 monocytes after the uptaking the MSU.

Abstract:Aim To study the effect of united therapy of probucol(P),clopidogrel(Anti-thrombosis,A) and atorvastatin(statin,S) on Markers of inflammation of rabbit atherogenesis. Methods 36 rabbits were randomized into four groups. One group did not receive treatments and served as control group. Atherosclerosis was induced in the aorta arteries of rabbits by atherogenic diet in the rest groups for 6 weeks. Then,animals were randomized to receive treatment or no treatment. One group of treatment were received atorvastatin,and the other accepted probucol and clopidogre and atorvastatin and killed after 4 weeks. Results Arterial macrophage infiltration was abolished by the treatment of atorvastatin and united group. Vascular cell adhesion molecule-1(VCAM-1) and interleukin-6(IL-6)were significantly diminished in the neointima and the media,and the same were monocyte chemoattractant protein-1(MCP-1). Notedly,there was a more evident effect in united therapy than Atorvastatin in anti-inflammation. Conclusions Atorvastatin and united therapy diminishes the neointimal inflammation in rabbit atherosclerosis model,and the effect of united therapy was more than atorvastatin. This could contribute to the more stabilization of the atherosclerotic plaque,and may be a more meaningful additional prediction for the reduction of acute ischemic events in patients treated with united therapy.

Abstract:Aim To investigate the effect of pobucol on the prevention of restenosis after balloon angioplasty in hypercholesterolaemic rabbits,and to examine the expression of growth factor. Methods 32 white rabbits were randomly divided into high cholesterol diet group (n=16)and probucol group (n=16),after one week of diet,all rabbits were injured on carotid artery by balloon angioplasty. Carotid artery tissues were harvested at 8 weeks after injury by balloon. During 8 weeks the rabbits were fed with hypercholesterolaemic diet and probucol. The areas of lumen of blood vessel,intima and elastic fibers dying were observed by HE staining,and the insulin-like growth factor-1 receptor (IGF-1R) and vascular endothelial growth factor (VEGF) were examined by immunohistochemical method. Results Compared with the high cholesterol diet group,the lumen areas of the probucol group were larger (P<0.01),the intimal areas were smaller (P<0.05 ),and the expression of IGF-1R and VEGF significantly decreased (P<0.05). Conclusions Probucol can prevent the restenosis of rabbits' carotid artery from balloon angioplasty injury,and inhibit the expression of IGF-1R and VEGF.

Abstract:Aim To compare the efficacy of probucol and atorvastatin in treatment of coronary heart diseases (CHD) with hyperlipidemia. Methods 119 patients with hyperlipidemia and CHD were randomly divided into three groups,received probucol 0.5 g bid or atorvastatin 10 mg qd or two drugs together. After 12 weeks,the result of treadmill exercise ECG test,total cholesterol (TC),triglyceride (TG),high density lipoprotein cholesterol (HDLC),low density lipoprotein cholesterol (LDLC),oxidized low density lipoprotein (ox-LDL),superoxide dismutase (SOD),malondialdehyde (MDA),and other indicators were studied. Results The serum TC and LDLC levels were significantly lowered (P<0.01) in three groups; HDLC level significantly increased in atorvastatin group(P<0.01),but the ox-LDL,SOD and MDA did not change significantly. ox-LDL and MDA decreased significantly in probucol group (P<0.001),SOD increased (P<0.001),but HDLC decreased (P<0.01). ox-LDL and MDA decreased significantly (P<0.001),SOD increased (P<0.001),HDLC did not change significantly in combined group. In three groups,treadmill exercise ECG test positive rate reduced after treatment and the number of angina attack reduced within two weeks before the end of research,probucol group was more significant than atorvastatin group (P<0.05),the combined group was more significant than the other two groups (P<0.01). Conclusion Probucol is a safe and effective drug for treating hypolipidemic and reducing angina attack. Its combination with atorvastatin is safe,and the efficacy is more visible.

Abstract:Aim To give an overview of the field of antioxidation and restenosis prevention with probucol,and investigate their potential use for the prevention of atherosclerosis progression. Methods A meta-analysis was performed by RevMan 4.2.10. We searched PubMed (1966~2007.9),EMBASE (1980~ 2007.9),CBMDisc(1980~2007.9),CNKI(1994~ 2007.9),the Cochrane Central Register of Controlled trials (CENTRAL,in the Cochrane Library -Issue 3,2007) and hand searched reference textbooks,articles and scientific proceedings. It was collected by Randomized controll trials (RCT) and quasi-RCT to compare probucol with placebo and no treatment or other convention. Two reviewers independently assessed trial quality and extracted data. Statistical analyses were performed by using the fixed or random effects model after testing for heterogeneity. The results were expressed as weighted mean difference (WMD) or standardized mean difference (SMD) for continuous outcomes and relative risk (RR) for dichotomous outcomes with 95% confidence intervals (CI). Results Seven studies involving 525 participants were identified for oxidized LDL levels. Average ox-LDL decreased significantly with probucol compared to placebo in all [SMD-2.38,95%CI (-3.39~-1.38)]. Four studies involving 179 participants were identified for flow-mediated vasodilation (FMD). Average FMD decreased significantly with probucol compared to placebo in all [SMD 1.97,95%CI (0.60~3.35)]. Four studies involving 335 participants were identified for lumen area (LA). Average LA decreased significantly with probucol compared to placebo in all [WMD 0.80,95%CI (0.41~1.20)]. Similarly,eleven studies involving 856 participants were identified for minimal lumen diameter (MLD). Average MLD decreased significantly with probucol compared to placebo in all [WMD 0.32,95%CI (0.22~0.42)]. The pooled result of 13 studies involving 1132 participants showed

Abstract:Aim To study the expression of hypoxia-inducible factor-1α(HIF-1α) in myocardial infarction rats and the role of protein kinase C (PKC) in signal pathway of hypoxia-inducible factor-1α gene expression. Methods 32 healthy male Wistar rats were divided into four groups:ischemic preconditioning group,simple myocardial infarction group,ischemic preconditioning plus protein kinase C inhibitor group and sham-operation group. Rats in treated groups received ligation of the proximal left anterior descending coronary artery to induce acute myocardial infarction. The sham-operation group dont take ligation of coronary artery. In ischemic preconditioning plus PKC inhibiter group,rats were injected with the protein kinase C inhibitor (chelerythrine,5 mg/kg,iv). After 1 day of operation,rats were killed. Results The infarct size in ischemic preconditioning group decreased significantly than that in ischemic preconditioning plus protein kinase C inhibitor group and simple myocardial infarction group (P<0.01). The hypoxia-inducible factor-1α mRNA gene expression increased in ischemic preconditioning group more than that in ischemic preconditioning plus protein kinase C inhibitor group and simple myocardial infarction group. Hypoxia-inducible factor-1α protein expression had the same change. Conclusions Hypoxia-inducible factor-1α play aprotective role in ischemic preconditioning; protein kinase C is the important signal pathway of hypoxia-inducible factor-1α gene expression.

Abstract:Aim To investigate the effects of celastrol on the collagen macrophage migration inhibitory factor and matrix metalloproteinase-9 expressions in the plaque of apoE gene knockout mice gene knockout mice. Methods 8-week old apoE gene knockout mice,male ,were divided randomly into atherosclerosis model group and celastrol treantment group( n=6 in each group). 8-week old male C57BL/6J mice were the normal control. The mice in celastrol group were given. celastrol 2 mg/(kg·d) by intraperitoneal injection for 4 weeks; the mice in control group were only given equivalent amount of dimethyl sulfoxide (DMSO),and the contents of collagen in the aortic atherosclerotic lesions were detected with Picrosirius Red staining. The expression of macrophage migration inhibitory factor and MMP-9 were detected by immunological histochemical method. Results The area of lipid plaque in the mice treated with celastrol was 4270.74±1027.64 μm2 ,significantly smaller than that of the model group 8971.19±1665.76 μm2 (P<0.01). The contents of collagen in the atherosclerotic plaque was significantly higher in Celastrol group compared with model group (mean optical density:0.0275±0.0068 vs 0.0142±0.0054,P<0.01). The expression of matrix metalloproteinase-9 was significantly decreased in celastrol group compared with the model group(mean optical density:0.0054±0.0020 vs 0.0263±0.0080,P<0. 001). The expression of macrophage migration inhibitory factor was also significantly decreased in celastrol group compared with the model tgroup (mean optical density:0.0114±0.0016 vs 0.0227±0.0039,P<0. 001). Conclusions Celastrol can inhibit the progress of atherosclerotic plaque and promote the stability of ather-osc1 erotic plaque in apoE gene knockout mice ,which might be relative to its inhibition on the degradation of collagen and expression of macrophage migration inhibitory factor,matrix metalloproteinase-9 in atherosclerotic plaque of apoE gene knockout mice.

Abstract:Aim To explore the effect of chloride channel blocker DIDS on cell signaling pathway phosphatidylinositol 3'-kinase/proteinase B (PI3K/Akt) and its downstream molecules endothelial nitric oxide synthase (eNOS) and nitric oxide (NO) in staurosporine (STS)-treated cardiomyocyte apoptosis. Methods Neonatal rat cardiomyocytes were exposed to STS in the presence or Absence of DIDS. Cell viability,apoptosis and expressions of Akt,phospho-Akt (p-Akt),eNOS,phospho-eNOS (p-eNOS) and NO production were determined. Results DIDS markedly improved cell viability on STS-exposed cardiomyocytes. DIDS resulted in a 2.1-fold increase of p-Akt over control levels,prevented the reduction in eNOS expression and phospho-eNOS levels induced by STS and significantly increased NO production (all P<0.01). Pretreatment with LY294002,a selective PI3K inhibitor,abolished DIDS-induced increases in p-Akt,eNOS,p-eNOS and NO production,and completely abrogated the DIDS-induced anti-apoptotic effect (P<0.01). Pretreatment with L-NAME,a non-selective NOS inhibitor similarly inhibited the increased NO but only partly abolished protective effects of DIDS (P<0.05). Conclusion DIDS inhibits STS-induced cardiomyocyte apoptosis via activating PI3K/Akt signaling pathway.

Abstract:Aim To observe the morphological changes in vascular aging-related remodeling,and p16INK4a,p21cip1 gene expression associated with senescence in healthy rats during natural aging. Methods Aortic structure and microstructure changes of vascular endothelial cell in 4,10,16,24 months old rats were observed respectively,and the expressions of p16INK4a,p21cip1 protein were determined by Western-Blotting in the same periods. Results With aging,aorta wall thickened,fibrosis degree increased,endothelial cells appeared flattened and enlarged. The protein expressions of p16INK4a,p21cip1 were increased in vascular aging-related remodeling,which suggests that p16INK4a,p21cip1 activity may play an important role in regulating vascular senescence lifespan in vitro. Conclusion Vascular aging has its specific structural alterations,one of its molecular mechanisms might be associated with increasing the expression level of p16INK4a,p21cip1 with aging. Further elucidating its underlying mechanism may provide theoretical basis for prevention and treatment of atherosclerosis.

Abstract:Aim To examine the effects of mesenchymal stem cells (MSC) intramyocardial injection after infarction on extracellular collagen matrix (ECCM),angiogenesis in ischemic myocardium and left ventricular function in ischemia-reperfused rats. Methods We created infarction in rat by ligaturing coronary anterior descending branch for 60 min and reperfused,administered DAPI-labeled allogeneic bone MSC intramyocardial injection before reperfusion and measured vessel density,cross-sectional area of viable myocardium,indexes of expansion and left ventricular remodeling,collagen volume fraction,and hemodynamic parameters at 7 and 28 days post-surgery. Results The infarct size were reduced and indexes of remodeling attenuated significantly,but without apparent improvement of ventricular function at 28 days post MSC injection. DAPI was found in infarct,and peri-infarct myocardium,some of them were vWF and α-SMA positive,which suggested that MSC might differentiate into smooth muscle cells and endothelial cells,result in increasing the vessel density in periphery region of infarction and reducing collagen profile. Conclusions In a ischemia-reperfusion rat MI model,MSC myocardial transplanted differentiated into vessel smooth muscle and endothelial cells,increased vascularity and therefore reduced infarct size and ECM,but without improving cardiac function.

Abstract:Aim To investigate the effect of calcitonin gene-related peptide (CGRP) on the proliferation and energy metabolism of vascular smooth muscle cells(VSMC) induced by angiotensin Ⅱ (AngⅡ),and further explore whether AMP-activated protein kinase (AMPK) involved in the cellular signal pathway. Methods VSMC were prepared from thoracic aorta of male Sprague-Dawley rat by the explanted method. The 3 ~ 10 passages of cells were used for the present studies. The viability of cultured VSMC was estimated by MTT assay and cells counting. Cellular energy metabolism was tested by ATP assay kit,Janus Green B staining. Western Blotting was used to observe the expressions of phospho-AMPK (p-AMPK) in VSMC. Results AngⅡ (100 nmol/L) increased the viability of VSMC. Pretreatment of the cell with CGRP significantly inhibited VSMC proliferation,hypertrophy of mitochondria,extra-generation of ATP induced by AngⅡ (P<0.05). Meanwhile,CGRP down-regulated expressions of p-AMPK induced by AngⅡ,which were partly abolished by CGRP8-37 or PD98059,respectively. Conclusion Pretreatment with CGRP inhibits the proliferation and energy metabolism of VSMC induced by AngⅡ. The signal transduction involving in this process is likely to be related with the signal linkage,namely,CGRP/CGRP receptor-1(CGRPR-1)-mitogen extracellular kinase (MEK)/ ERK1/2-p-AMPK.

Abstract:Aim To compare the adhesion and proliferation of late endothelial progenitor cells (EPC) isolated and expanded from rabbit peripheral blood cultured with nanofibers and alignate nanofibers synthetic poly-L-lactic acid (PLLA) scaffold in vitro for applications in vascular tissue engineering,thus to optimize PLLA scaffold materials. Methods Nanofibrous scaffolds of PLLA by electrospinning were modified by hypothermy plasma and type I collagen was coated onto the materials physically. In vitro,late EPC were cultured on the PLLA scaffold. Furthermore,cell incorporation of DiI-labelled ac-LDL and binding of FITC-UEA-I was determined by fluorescent microscopy. Adherency and proliferation of EPC in two groups at different time points were surveyed and to observe morphological change of late EPC on PLLA scaffold was observed by contrast phase microscope,fluorescent microscope and scanning electron microscope. Results The nanofibrous scaffolds consists of fibers with diameters ranging from 300 nm to 400 nm,whose porosities were more than 90%. The adherency of late EPC on scaffold were completed in 12 hours. The proliferation of late EPC on alignate nanofibers scaffold was markedly increased compared to control groups in every time (P<0.05). Late EPC grew well with PLLA scaffold,yet confused and disorderly in control groups. Late EPC could attach,extend,proliferate and secrete extracellular matrix following fibrous orientation in alignate nanofibers groups of PLLA,which majority of the fibers were oriented along the longitudinal axis which form a unique aligned topography. Conclusions Late EPC are ideal seeding cells for tissue engineering. Late EPC can adhere well to alignate nanofibers of PLLA and proliferate,keep well on cell morphology. This type of PLLA scallfold is a optimal candidate material for late EPC transplantation in vascular tissue engineering.

Abstract:Aim To observe the level of plasma vascular endothelial growth factor (VEGF),stromal cell-derived factor-1α (SDF-1α),whether mobilization of endothelial progenitor cells (EPC) is impaired in patients of acute myocardial infarction (AMI) with type 2 diabetics,and the pathway of SDF-1α,VEGF-EPC is abnormal. Methods Circulating CD45-/low+/ CD34+/ CD133+/ KDR+ early EPC count in peripheral blood mononuclear cells(PBMC) were quantified by flow cytometric analysis on day 1,3,5,7,14,28 after AMI. Plasma VEGF,SDF-1α and high sensitivity C-reactive protein (hs-CRP) level were mearsured at the above time points as those at EPC number counts with a standardized ELISA-kit. Results In non-diabetic patients,circulating EPC count increased after AMI,with the highest peak at day 5,following which EPC count gradually returned to baseline. In patients with type 2 diabetes,the highest peak was delayed (day 7 vs day 5),and the magnitude of EPC mobilization was decreased as compared with non-diabetic subjects [highest EPC count:(140±48)/106 vs (246±100)/106,P<0.05]. Plasma VEGF [day 5:277±95 ng/L vs 168±35 ng/L,P<0.05],SDF-1α [day 5:3 835±402 ng/L vs 3 287±384 ng/L,P<0.05]and hs-CRP[day 3:55.55±14.88 mg/L vs 36.92±14.83 mg/L,P<0.05] were significantly higher in AMI patients with diabetics than those without diabetics. Conclusion Tissue ischemia is more serious and ischemia-induced bone marrow-derived EPC mobilization is impaired in AMI patients with type 2 diabetics,and such impairment is likely due to impaired SDF-1α,VEGF -EPC mobilization pathway. This abnormal SDF-1α,VEGF-EPC mobilization pathway possibly contributes to the poor collateralization observed in diabetic patients in response to vascular occlusive disease.

Abstract:Aim To investigate the association between coronary heart disease (CHD) and heme oxygenase-1 (HO-1) promoter region (GT)n repeated sequence polymorphism and the influence of the level of serum bilirubin. Methods The polymorphism of HO-1 gene in 337 CHD patients and 240 health controls was analyzed by polymerase chain reaction-native polyacrylamide gel electrophoresis,and the level of serum bilirubin of each patient at the same time was detected. Results The HO-1 genotypes were classified into three groups:LL,LS and SS. The LL genotypic frequency and L allele frequency in CHD patients was significantly higher than control group (P<0.05); the L allele frequency had different distribution in single,double and multivessel disease (P<0.05),and the LL genotypic frequency of multivessel disease patients was higher than that of single and double vessel disease patients (P<0.05); bilirubin level of LL genotype patients was significantly lower than that of SS genotype patients (P<0.001). Conclusions The polymorphism of HO-1 promoter region (GT)n repeated sequence plays an important role in CHD pathobiology process,L allele frequency is a risk factor of CHD,which maybe correlate with its potential ability to decrease the serum bilirubin.