Abstract:Aim To examine whether procucol could inhibit vascular smooth muscle cells(VSMC) proliferation by impairing cell cycle progression. Methods Initially,quiescent rat VSMC were treated with probucol in the presence of oxidized low density lipoprotein(ox-LDL).Then cell proliferation was measured by MTT assay and cell counts,and cell cycle distribution was analyzed by Flow-cytometry(FCM).Finally,protein expression was examined by western blot. Results MTT assay and cell counts revealed that 25 mg/L,50 mg/L and 100 mg/L ox-LDL stimulated VSMC proliferation in a dose dependent manner.Probucol significantly inhibited VSMC proliferation induced by ox-LDL in a dose and time dependent manner.FCM analysis showed that probucol increased cells in G₀/G₁ by 28.6%(p<0.01,n=3),reduced the cells in S by 83.5%(p<0.01,n=3).Also,100 mg/L oxLDL induced cyclin D1 expression,with peak in 6～12 h.Probucol markedly inhibited cyclin D1 expression by 26% at 6 h and 23% at 12 h respectively(both p<0.01,n=3). Conlusions Probucol significantly inhibits ox-LDL induced VSMC proliferation.This antiproliferative effect of probucol is correlated with suppression of cell progression by blocking the cell cycle in G₀/G₁.Furthermore,suppression of cell cycle progression is associated with the down-expression of cyclin D_1 protein by probucol

Abstract:Aim To investigate the effects of atorvastatin on AngⅡ-induced hypertrophy myocytes and the changes of mRNA expression of peroxisome proliferators-activated receptorβ/δ in vitro. Methods Hypertrophy in neonatal rat cardiac myocytes(MC) was established with angiotensin Ⅱ(Ang Ⅱ) and treated with atorvastatin.The surface area of MC was analyzed by the aid of NIH Image J software,and the synthetic rate of protein in MC was detected by 3H-leucine incorporation.mRNA expression of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP) and peroxisome proliferatoractivated receptor beta/delta subtypes(PPARβ/δ) was measured by reverse transcription-polymerase chain reaction(RT-PCR). Results In the condition of hypertrophy,increases of surface area(p<0.01),mRNA expression of ANP,BNP(both p<0.01),and ~3H-leucine incorporation(p<0.01);and a decrease of mRNA expression of PPARβ/δ(p<0.01) in MC were detected,but no changes in treated with DMSO(p>0.05).Atorvatatin inhibited the changes above,reduced mRNA expression of ANP and BNP,elevated mRNA expression of PPARβ/δ in a dose-dependent manner(p<0.05). Conclusions It was suggested that atorvastatin has a potential role in the prevention and treatment of cardiac hypertrophy,and PPAR β/δ may be involved in it.

Abstract:Aim To study the role of acylation stimulating protein for the differentiation of 3T3-F442A preadipocytes and explore the molecular mechanism of it. Methods There were two groups in this study: acylation stimulating protein (AsP) group(induced to differentiate by AsP) and control group(without inducers).The morphological changes were observed by Oil-Red O staining.Triglyceride(TG) synthesis and TG mass were assayed by chemical colorimetry. The cell were harvested on the 0~(th) day,1~(th) day,2~(th), 4~(th) day,6~(th) day after induced differentiation and total RNA of these cell were abstracted. The transcription factor peroxisome proliferator activated receptor γ(PPARγ), C/EBPα,C/EBPδ mRNAs expression were assayed by reverse transcription-polymerase chain reaction(RT-PCR). Results 3T3-F442A preadipocytes were induced to differentiate with AsP alone.Fat droplets were clearly visible in the cytoplasm of 3T3-F442A cell.The differentiation rate were high(80%) in AsP group. In AsP group TG synthesis and TG mass were significantly increased,both of them were higher than that of control group(p<0.05). PPARγ mRNA expression in the 3T3-F442A cell were low on 0~(th) day of induced differentiation,and it was increase significantly on the 1~(th) and 2~(th) day induced by AsP(p<0.05). It continued to increase significantly on the 4~(th) and 6~(th) day(p<0.05).The C/EBPδ mRNA were expressed in low level on 0~(th) day of differentiation induced by AsP and the expression level was increase significantly on the1~(th) and 2~(th) day of differentiation(p<0.05).But the expression of C/EBPδ mRNA was decreased on the 4~(th) and 6~(th) day,there were no significant difference compared with that of 0th day(p>0.05).The expression of C/EBPα mRNA was very low on the 0~(th) day of differentiation induced by AsP.It was increased on the 1~(th) and 2~(th)

Abstract:Aim To acquaint with atherosclerosis(As) progress of apolipoprotein E~-knockout(ApoE~-) mouse in different periods and explore the influence of different diets on As progressof ApoE~-mice. Methods 40 ApoE~-mice with 6 week age,20 mice among them were given west food diet and 20 mice were given common diet,4 mice were randomly chosen in each group at the period of 15,19,24,28 and 36 weeks and sacrificed for pathological determination. Results As of ApoE~-mouse gradually aggravated with time passing, undergone a typical pathological course from fatty streak to atheromatous plaque;As of ApoE~-mice with high fat diet were more severe than As of ApoE~-mouse with common diet. Conclusions ApoE~-mouse was a reliable model for As research;west food diet could accelerate As pathological progress of ApoE~-mouse.

Abstract:Aim To testify that nitric oxide(NO)/ cGMP-dependent protein kinase(PKG) can regulate expression of inositol 1,4,5-trisphosphate receptor type Ⅰ(IP3RⅠ) and further adjust intracellular Ca²⁺ level in vascular smooth muscle cells(VSMC). Methods Colorimetric Assay(MTT assay) was used for cellular growth.Fluorescence measurements of [Ca²⁺]i were performed using a laserscanning confocal microscope.IP3RI expression was determined by using reverse transcripase-polymerase chain reaction(RT-PCR) and immunoblotting methods. Results Proliferative response of VSMC was inhibited by S-nitroso-N-acetylpenicillamine(SNAP) and Sp8-pCPT-cGMP and promoted by Rp-8-pCPT-cGMP.After VSMC were pretreated with Verapamil,cell proliferation was further inhibited by SNAP and Sp-8pCPT-cGMP respectively.Verapamil inhibited phenylephrine(PE)-stimulated intracellular Ca²⁺ variations,which could be further inhibited by SNAP and Sp-8pCPT-cGMP respectively and slightly promoted by Rp-8-pCPT-cGMP.IP3R Ⅰ mRNA expression levels were decreased by SNAP and Sp-8-pCPT-cGMP,and increased by Rp-8-pCPT-cGMP.Protein levels of IP3R Ⅰ paralleled the changes in mRNA levels in response to preceding stimulus factors. Conclusion NO/PKG can suppress IP3R Ⅰ expression at the transcriptional and translational level,and further modulate intracellular Ca²⁺ concentrations in VSMC.

Abstract:Aim To explore the influence of endothelin-1(ET-1) on the gene expression of human vascular endothelial cells and the molecular mechanism of vascular endothelial cells injury induced by ET-1. Methods ECV304 cells were cultured and the total RNA was extracted.The GEArry Q Series Human Endothelial Cell Biology Gene Array was used to analyze the differential expression of genes associated with the major functions of endothelial cells 12 hours after treated with ET-1. Results Compared with control group,expression of 53 genes was differentiated at the time point of 12-hour in endothelin group.Among them,expression of 22 genes including mainly antithrombotic and antioxidant genes was downregulated and that of 31 genes including mainly prothrombotic and inflammatory factor genes was upregulated. Conclusions ET-1 could injure human vascular endothelial cells by downregulating antithrombotic and antioxidant genes and upregulating prothrombotic and inflammatory factors genes

Abstract:Aim To observe the expression change of resistin,adiponectin and adiponectin receptor of adipose tissue in high fat-fed Wistar rats,and to clarify the action of resistin,adiponectin with its receptor on the development of insulin resistance. Methods 30 Wistar male rats were randomly distributed into 2 groups: control and high fat-fed group.After they were treated for 11 weeks, total RNA,was extracted,reverse transcription(RT),polymerase chain reaction(PCR) was executed and enlarged the isolated cDNA and half-quantitative analysis was used. Results In high fat-fed group weight increased obviously than control group,as well as fasting blood glucose,low density lipoprotein cholesterol(LDLC), total free fat acid,insulin and HOMA-insulin resistance(IR),while HDLC(high density lipoprotein cholesterol) decreased(p<0.01).The glucose and insulin tolerance reduced greatly(p<0.01);the expression of adiponectin and resistin of high fat-fed group dropped markedly(p<0.01),whereas the expression of adiponectin receptor 1(R1) only showed the tendency to reduce,but didn't achieve significant statistic differences. Conclusions The decrease expression of adiponectin and resistin play an important role in the high fat-fed induced insulin resistance.Reduced expression of R1 may possibly associate the resistance of adiponectin with the high fat-fed induced insulin resistance.

Abstract:Aim To observe the effects of Tongmai granules on blood lipids,aorta and coronary artery pathological morphology in atherosclerosis rabbit model. Methods Forty Japanese big ear rabbits were randomly divided into negative control group,model control group,low dosage TongMai granules group and high dosage TongMai granules group,each group consisting of ten rabbits.The negative control group was fed on common diet,the other groups were high-lipid fed(5% pork fat,0.5% cholesterol) to establish atherosclerosis model.The latter two groups were given low dosage and high dosage Tongmai granules respectively.Blood lipid,aorta and coronary artery pathological morphology(including the thickness and the area of the aorta atherosclerosis plaque and the percentage of stenosis of the coronary artery supplying left ventricle) were observed in all the rabbits before and/or after the treatment. Results Blood TG,TC,LDLC were elevated significantly in the rabbits of the model group,but the elevation in the Tongmai group was lower than those of the model group,especially in the high dosage group.There are significant differences as compared with those of other groups(p<0.05).Normal aorta and coronary arteries are seen in the negative control group while in the other groups atherosclerosis of various degrees and coronary arteries stenosis came into being.Aorta plaque thickness,plague area/ aorta area and the percentage of stenosis of coronary arteries is lower in the Tongmai group than those in the model group, with those of the high dosage group being the lowest(p<0.05). Conclusions Tongmai granules can significantly reduce blood lipids,prevent and treat atherosclerosis.The mechanisms by which Tongmai granules alleviates angina pectoris may be attributed to its properties of reducing blood lipids,preventing and treating atherosclerosis

Abstract:Aim To investigate the effects of 10-23DNA enzyme(ED5) on endothelial function and ultrastructure change of Carotid Artery after injured artery in rats. Methods Ninety-six rats were randomly divided into sham-group,control-group 1,control-group 2 and ED5-group(n=24).The models of endothelial denudation of rats were made by using balloon catheter,FITC-ED5 was delivered to the artery wall of ED5-group in a total of 200 μL solution.Rats of control-group1 were given 200 μL 1 mmol/L MgCl2.Rats of control-group2 were given FuGENE6 Reagent.Six rats were killed at the 3rd,7th,14th,21th day after balloon injury.The serum level of nitric oxide and nitric oxide synthase and endothelin were detected.At the same time,to observe pathological change of injured arteries by optical microscope and electron microscope. Results The serum level of nitric oxide and nitric oxide synthase in ED5-group were higher than those in 2 control groups,increased significantly on the 14th day(nitric oxide: 57.1±1.9 μmol/L vs 38.7±1.9 μmol/L and 57.1±1.9 μmol/L vs 38.3±1.9 μmol/L,p<0.05;nitric oxide synthase: 11.1±0.4 μmol/L vs 8.1±0.4 μmol/L and 11.1±0.4 μmol/L vs 8.0±0.4 μmol/L,p<0.05);but endothelin in ED5-group was lower than that in 2 control groups,obviously different on the 14th day(111.2±7.2 pg/L vs 136.6±7.2 pg/L and 111.2±7.2 pg/L vs 135.5±7.2 pg/L,p<0.05).Compared with ED5-group,the neointimal thickness in 2 control groups increased significantly on the 21th day after balloon injury(64.0±4.2 μm vs 81.1±4.9 μm and 64.0±4.2 μm vs 79.5±3.9 μm, p<0.01).The vascular smooth muscle cell of neointima in 2 control groups may be contractile type,but vascular smooth muscle cell of neointima in ED5-group may be synthetic type. Conclusion The ED5 can improve the endothelial function after artery injury and inhibit the phenotype transform of vascular smooth muscle cells,and reduce the degree of neointimal hyperplasia.

Abstract:Aim To investigate effects of Propolis-ethanol extract on platelet adhesion to collagen function. Methods Intima-injury model was copied by using vitro perfusion.Platelet coverage on collegen was measured when shearing stress was 1000/s after blood passing collagen surface.We set three groups:negative control group(24% ethanol),experimental group(0.1 g/L,24% Propolis-ethanol extract),positive control group(0.1 g/L,ferulic acid). Results Platelet coverage in experimental group was decreased from 24.0% to 11.5%,there was significant difference than that in negative control group(p<0.01);There was no significant difference between experimental group and positive control group(p>0.05). Conclusions Propolis-ethanol extract could decrease the platelet adhesiveness area on collagen surface.

Abstract:Aim To screen the mutations in lipoprotein lipase(LPL) gene exon 9 and to investigate the effect of mutation on triglyceride(TG) and high density lipoprotein cholesterol(HDLC) level. Methods The exon 9 of LPL gene was amplified by PCR and the mutations were examined by polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP);the natures of mutations were identified by DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). Results Only nonsense mutation Ser~(447)→stop was screened by PCR-SSCP,one subject was homozygous and 77 subjects were heterozygous.The frequency of stop~(447) allele is 9.4% and the incidence of Ser~(447)→stop was(18.6%) in the 420 subjects with normal TG level.The triglyceride level in LPL stop~(447) carriers(1.05±0.32 mmol/L)was lower than that in LPL stop~(447)non-carriers(1.13±0.28 mmol/L)(p<0.05);HDLC level in the former(1.28±0.28 mmol/L)was higher than the latter(1.25±0.27 mmol/L)(p> 0.05). Conclusions Only Ser~(447)→stop mutation exists in LPL exon 9 of 420 subjects with normal TG and the Ser~(447)→stop mutation have the minor effect on lowering TG level.

Abstract:Aim The effect of crocetin on relaxation function of thoracic aorta isolated from hyperlipidemic rabbits was examined and the potential mechanism was explored. Methods Hyperlipidemic rabbit model was established by feeding high lipid diet (HLD) to rabbit.Blood was collected at week 0,2,4,6,8 respectively and serum was used for measurement of nitric oxide(NO) content.At the end of 8th week,serum total cholesterol(TC) and low density lipoprotein cholesterol(LDLC) contents were assayed and thoracic aorta isolated from the rabbit was mounted in an organ bath to measure the endothelium-dependent and-independent relaxations evoked by acetylcholine(Ach) and sodium nitroprusside(SNP) respectively.NO synthase(NOS) activities and mRNA expression of endothelial NO synthase(eNOS) of aorta was determined. Results Endothelium-dependent relaxation of thoracic aorta evoked by Ach was found significantly impaired in merely HLD-treated rabbits with the maximal relaxation of 54% of the control.This impairment was significantly improved by co-treatment with CCT(15,30 mg/kg),with a maximal relaxation of 68% and 80% of the control respectively.Endothelium-independent relaxation induced by SNP maintained comparable in all groups.Compared with HLD group,serum levels of TC and LDLC were decreased in CCT(30 mg/kg) group by 21.6% and 20.2% respectively rather than in CCT(15 mg/kg) group.CCT(15,30 mg/kg) could increase serum NO content by 36.1% and 72.4% respectively at the end of 8th week.Endthelial NOS activity was significantly increased by CCT(30,15 mg/kg) by 76.1% and 47.8% respectively and mRNA expression of eNOS increased by 54.2% and 29.8% respectively.Inducible NOS(iNOS) activity remained unchangeable in all groups. Conclusion CCT could significantly increase the serum NO content and restore endothelium-dependent relaxation in thoracic aorta isolated from hyperlipidemic rabbit.The mechanism under which might be related to increased mRNA expression of vessel eNOS

Abstract:Aim To investigate effects of L-arginine and N~G-nitro-L-arginine methyl ester(L-NAME) on human umbilical vein endothelial cell(hUVEC) secreting nitric oxide(NO) and superoxide anion(O_2~-). Methods hUVEC was incubated with glucose,insulin,L-arginine and L-NAME respectively for 24 hours.The activity of nitric oxide synthase(NOS) and superoxide dismutase(SOD),the concentration of NO and O_2~-in the supernatant of cultured hUVEC was measured at the end of the experiment. Results NOS activity,the concentration of NO increased,while SOD activity decreased and the concentration of O_2~-increased significantly in the presence of 25 mmol/L glucose.NOS,NO,SOD did not change significantly in the presence of L-arginine,while the concentration of O_2~-decreased significantly. The activity of NOS,the concentration of NO increased in the presence of 25 mmol/L glucose+L-arginine,L-arginine may reverse the elevation of O_2~-which induced by glucose.NOS,NO increased in the presence of 10,100,1 000 mu/L insulin,while SOD and O_2~-did not change significantly.NOS,NO increased in the presence of L-arginine+insulin,SOD activity did not change significantly while O_2~-decreased.NOS activity and the generation of NO decreased in the presence of 100 μmol/L L-NAME,while SOD activity did not change significantly,however the generation of O_2~-increased.NOS,NO decreased in the presence of 25 mmol/L glucose+L-NAME,however SOD activity and the concentration of O_2~-did not change significantly.NOS,NO decreased in the presence of 10 mu insulin+10 μmol/L LNAME and 100 mu insulin+100 μmol/L L-NAME,SOD activity did not change significantly,while the concentration of O_2~-increased. Conclusion NOS,NO,SOD did not change significantly while O_2~-decreased in the presence of L-arginine.NOS activity,the generation of NO decreased while O_2~-increased in the presence of L-NAME in the supernatant of cultured endothelial cell.

Abstract:Aim To investigate the effect of 17β-estradiol(17β-E₂) eluting stent implantation on neointima proliferation and re-endothelialization of abdominal aorta in rabbits. Methods Male rabbits were divided into 3 groups after hyperlipemia feed.17β-E₂ eluting stents,phosphorylcholine(PC) coated metal stents and bare metal stents were implanted in each groups.The stented aortas tissue of 6 rabbits were removed at 2 weeks,4 weeks and 12 weeks from each group respectively.Lumen area,thickness and area of neointima at different time after stent implantation were measured by computer image analysis technique.Immunohistochemistry was used to detect the positive rate of factor Ⅷ expression of the arterial intima at different time in order to assess re-endothelialization degree. Results The mean injury scores were similar in three groups(2.17±0.22,(2.18)±0.21 and 2.17±0.19,p>0.05).The neointima areas had increased at 2 weeks after stent implantation,and the lumen loss rates were less than 50% at 12 weeks in three groups.At 12 weeks neointima area reduced 36% in 17β-E₂ eluting stent group compared with that of bare metal stent group,and had no difference between PC coated and bare metal stent groups.The ratio of neointima area at the time of 2 week and 12 week was compared within each group,and the results showed that the 17β-E₂ group had significantly higher ratio than other two groups(0.77,0.61 and 0.61,respectively,p<0.01).At the time of 2 week the re-endothelialization rate in 17β-E₂ eluting stent group was obviously higher than PC coated stent and bare metal stent groups((78.4%)±2.3% vs 61.4%±3.4% and 62.8%±2.9%,p<0.01),and nearly complete re-endothelialization were found at 4 weeks after stent implantation procedure in three groups. Conclusions 17β-E₂ eluting stent has ideal effect in preventing restenosis by inhibition of neointima proliferation as well as acceleration of re-endothelialization of treated arteries.

Abstract:Aim To observe the effects of propranolol on peripheral blood leucocyte CD11a expression of experimental hyperlipemia rabbits and the formation of atherosclerosis. Methods Twenty-one male rabbits were divided into three groups: normal group,model group and propranolol group[5 mg/(kg.d)]. Rabbits of last two groups were fed on 0.5% cholesterol and 5% lard diet for 16 weeks and were injected with 10% bovine serum albumin.Peripheral blood adhesion molecule CD11a were detected with flow cytometry. The aortas were harvested for histomorphometric observation and quantitative analysis. The ratio of atherosclerotic plaque area to intimal area and content of macrophage were detected. Results Compared with model group,propranolol group showed that the lesion degree of atherosclerotic plaque lessened significantly and normal group were not found the formation of atheroslerosis.In propranolol group,the ratio of atherosclerotic plaque area to intima area and content of macrophage and the positive percentage of peripheral blood leucocyte CD11a expression were 59.1%±11.3%,56.3%±4.9%,61.9%±4.2% respectively. In model group,they were 78.3%±6.7%,70.6%±5.6%,74.3%±3.6% respectively,propranolol group decreased significantly(p<0.05). Conclusion Propranolol can postpone the formation of atherosclerosis through lessening the inflammation in the lesion of atherosclerosis of rabbits and increase the stability of atherosclerotic plaque.

Abstract:Aim To investigate the effect of endothelin-1(ET-1) on osteopontin expression of rat aortic vascular smooth muscle cell(VSMC). Methods Rat aortic vascular smooth muscle cell were cultured in vitro with DMEM and 10 percent calf plasma.The cell were incubated with different concentration of ET-1 for different hours,then RT-PCR and Western bloting analysis were used to observe the effect of H₂O₂ on osteopontin expression of rat VSMC. Results Different concentration of ET-1 could obviously induce gene expression and protein synthesis of osteopontin,without exhibiting dose-dependent promotion effect.Cell incubated with ET-1 for different hours also obviously induce gene expression and protein synthesis of osteopontin except 6 h,and exhibited time-dependence. Conclusions These results demonstrate that ET-1 could stimulate osteopontin expression of rat VSMC.

Abstract:Aim To examine the effect of epigallocatechin-3-gallate(EGCG) on intimal hyperplasia in a vein graft model by local delivery. Methods Rabbit jugular vein segments were incubated in saline containing EGCG 0.1 g/L.HPLC and FITC-labelled EGCG were used to assess the absorption characteristics of EGCG in vitro.Autogenous vein graft model was established in 20 rabbits which were randomly divided into two groups: EGCG group and control group.Vein grafts were incubated with EGCG or saline before arterial interposition grafting(n=10).Animals were sacrificed 21 days later and intimal thickening(intimal thickness and intimalto-medial ratio,I and I/M) was assessed by computerized digital morphometry.The proliferation and apoptosis of neointimal cells were determined by Ki67 staining and terminal deoxynucleotidyl transferase biotin nick end-labeling(TUNEL) method,respectively. Results At dose of 0.1 mg/L,the vein segments showed evidence of EGCG localization,EGCG was absorbed at levels of 2.9±0.9 mg/g,5.8±2.1 mg/g and 8.0±2.3 mg/g after incubation for 1 h,2 h and 4 h,respectively.All animals survived until the time of harvest,and there was 1 closed case respectively in two study groups.The EGCG group showed a significant reduction in neointimal formation at 21 days compared with the control condition(41.1±13.6 μm vs 89.9±48.3 μm,p<0.01 and 0.40±0.18 vs 0.77±0.31,p<0.05).Immunohistochemical analysis of Ki67 also indicated decreased rate of positive smooth muscle cells in the EGCG group(22.4%±8.6% vs 8.8%±(2.4%),p<0.05).Cell apoptosis was not different in two groups(0.40%±0.55% vs 0.60%±0.89%,p>0.05). Conclusions These results indicate that the local delivery of EGCG prevent intimal hyperplasia in a rabbit vein bypass grafting model through inhibition of neointimal smooth muscle cells proliferation.

Abstract:Aim To investigate the effects and potential mechanisms of atorvastatin on atherosclerotic lesion of hypercholesterolemic rabbits. Methods Fourteen male New Zealand rabbits were randomly divided into normal diet group(n=4),high-cholesterol diet group(n=10).After 8 weeks,highcholesterol diet rabbits were randomly switched to receive either atorvastatin 1.5 mg/(kg·d)(n=5) or starch(n=5).Two weeks later,the aortas of all rabbits were removed under deep anesthetization.The distal portion of aorta to the iliac bifurcation was exercised for hematoxylin-eosin staining and monocyte chemotactic protein-1(MCP-1) mRNA detection by in-situs hybridization. Atherosclerotic area,intima and media thickness were measured by experienced pathologist using Beihang pathology imaging analysis system.MCP-1 expression was expressed as average positive cell counts per ten HP. Plasma interleukin(IL-6) levels were determined by enzyme-linked immunosorbent assay(ELISA). Results Plasma IL-6 levels positively correlated with plasma low density lipoprotein cholesterol(LDLC) levels(r=0.852,p<0.01). Compared with starch group,atorvastatin treatment decreased plasma IL-6 levels by 55% and resulted in the decreased atherosclerotic area(52.5%±4.2% vs 81.9%±2.8%,p<0.01) and reduced intimae thickness(24.18±10.21 μm vs 77.51±22.47 μm,p<0.01).Additionally,atorvastatin treatment led to MCP1 positive cell counts decreased in aorta intimae(29±5 /HP vs 49±17 /HP,p<0.01).Intimae thickness of aorta was positively correlated with MCP-1 mRNA expression in intimae(r=0.831,p<0.01). Conclusion Hypercholesterolemia may induce systemic inflammation.Atorvastatin treatment can take anti-atherosclerotic effects possibly through reducing plasma cholesterol levels and inhibiting plasma IL-6 and MCP-1 expression in intima.

Abstract:Aim To observe the role of quercetin(QUE),isorhamnetin(ISOR) and norepinephrine(NE) on the proliferation of human aortic vascular smooth muscle cell(VSMC). Methods The effects of QUE,ISOR and NE on these VSMC growth and DNA synthesis was investigated by cell counting,~3 H-thymidine incorporation. Results QUE could inhibit proliferation and DNA synthesis of VSMC but these effects of ISOR was weaker than that of QUE.When VSMC were treated by 1～200 μmol/L of QUE or ISOR,the best inhibitory effects occured at 200 μmol/L.There were dose-dependent inhibitory effects in different concentration of QUE and ISOR.NE 10 μmol/L could stimulate proliferation and DNA synthesis of VSMC.The effects could be inhibited by phentolamine(Ph).QUE and ISOR markedly inhibited proliferation and DNA synthesis of VSMC induced by NE in a dose dependent manner.At the same time QUE and ISOR had cooperative effects on inhibition of the stimulation of NE.QUE and ISOR also had more powerful inhibitory effect on proliferation and DNA synthesis of VSMC co-incubation with NE than Ph.There were no cyto-toxic effects on VSMC treated with QUE and ISOR. Conclusion These results indicated that QUE and ISOR had no cyto-toxic effect on VSMC cultured in vitro and could effectively inhibit the proliferation and DNA synthesis of VSMC,especially inhibit the proliferation and DNA synthesis of VSMC stimulated by NE.

Abstract:Aim To study the effects of BPI-1096,a nitric oxide-releasing aspirin,on platelet aggregation. Methods Platelet-rich plasma(PRP) samples was prepared from healthy adult volunteers.The antiaggregant effects of BPI-1096 were tested by a standard nephelometric technique after incubating the drug with platelet at 37℃ for 10 min. Results The platelet aggregation induced by adenosine diphosphate(ADP),platelet activating factor(PAF),adnephrin and restocetin was inhibited by BPI-1096 at different concentration(p<0.05).BPI1096 inhibited platelet aggregation induced by arachidonic acid in every concentration but did ont show any significance(p>0.05).BPI-1096 displayed inhibitory effects similar to ASA at the same concentration.BPI-1096 did not strengthen the inhibitory effects on platelet aggregation at high concentration. Conclusions As a newly nitric oxide-releasing aspirin,BPI-1096 can significantly inhibit platelet aggregation induced by adnephrin,restocetin,ADP and PAF in vitro;the antiaggregation effects was similar to Aspirin at the same concentration.BPI-1096 did not show the correlation between concentration and efficiency.

Abstract:Aim To observe the change of thrombomodulin and its relationship with tumor necrosis factor-α(TNF-α)in acute cerebral infarct with hospital acquired pneumonia(HAP). Methods The patients with acute cerebral infarct were divided into pure infarct group(n=24),and acute infarct with HAP group(n=16),compared with normal control group(n=13).The serum samples were obtained from 1 d to 21 d after infarct.The serum level of thrombomodulin and TNF-α were examined by enzyme-linked immunosorbent assay(ELISA) method and radioimmunoassay respectively. Results The serum levels of thrombomodulin of acute infarct with HAP group were significantly lower than those of pure infarct group in the first week and 14 d after infarct(p<0.05),with similar result at 21 d.The serum levels of TNF-α of acute infarct with HAP group were significantly higher than those of pure infarct group from the first week to 21 d after infarct(p<0.05),although HAP had been prohibited in 21 d.In the group of acute infarct with HAP the serum levels of thrombomodulin were related inversely with that of TNF-α.The relationship between thrombomodulin and TNF-α compromised with Gompertz curve. Conclusions It's possible that the increase of serum TNF-α in acute infarct with HAP may injury the endotheliocyte of brain vessel and prohibited the anticoagulative system,presenting with downregulation of the level of serum thrombomodulin.

Abstract:Aim To investigate whether the plasma plasminogen activator inhibitor-1(PAI-1) activity and gene polymorphism played a pathogenetic role in acute coronary syndrome(ACS). Methods 106 patients suffering from acute coronary syndrome(ACS) and 98 subjects,who were suffering from stable coronary heart disease(SCHD),and 60 normal controls were recruited.The 4G/5G allele polymorphism were genotyped by using polymerase chain reaction technique,while plasma activity assayed by enzyme linked immunosorbent assay. Results The plasma PAI-1 activity level in the ACS group(18.0±2.9 kAU/L) was significantly higher than that in the SCHD group(16.8±2.7 kAU/L) and in the control group(16.2±2.8 kAU/L;p<0.01 or p<0.005).The frequency of 4G/4G allele homozygous in the ACS group(49.1%) was significantly higher than that in the SCHD group(28.6%) and in the control group(26.7%;p<0.05).The plasma PAI-1 activity level was statistically higher in 4G allele homozygous than those in 4G/5G heterozygous and 5G homozygous(p<0.05). Conclusions There may be a link between the plasma PAI-1 activity and it's 4G/5G genotype.Elevated plasma PAI-1 activity and essentially 4G allele homozygous genotype might be the major risk factors of ACS.

Abstract:Aim To study the relationships between carotid plaque and plasma matrix metalloproteinase and tissue inhibitor metalloproteinase in patients with acute coronary syndrome(ACS). Methods Using high-resolution ultrasonic instrument the intima-media thickness(IMT)and plaque score in carotid artery were detected in 30 patients with ACS,29 patients with stable angina pectoris(SAP)and 17 control subjects,and the plasma concentration of matrix metalloproteinase9(MMP-9)and tissue inhibitor metalloproteinase-1(TIMP-1)were measured by sandwich enzyme immunoassay technique. Results Compared with SAP group and control group,plasma MMP-9,TIMP-1,MMP-9/TIMP-1,and IMT,whole plaque score,soft plaque score and hard plaque score in carotid artery in ACS group were significantly increased(p<0.01).MMP-9 had positive relationship with IMT,whole plaque score and soft plaque score in carotid artery(r=0.468,p<0.01;r=0.592,p<0.01;r=0.677,p<0.01),respectively.TIMP-1 had positive relationship with IMT,whole plaque score,soft plaque score and hard plaque score in carotid artery(r=0.449,p<0.01;r=0.493,p<0.01;r=0.435,p<0.01;r=0.227,p<0.05),respectively.MMP-9/TIMP-1 had positive relationship with IMT,whole plaque score and soft plaque score in carotid artery(r=(0.253),p<0.05;r=0.431,p<0.01;r=0.547,p<0.01), respectively.Removing age factor,partial correlation analysis showed IMT had positive relationship with MMP-9(r=0.461,p<0.001),TIMP-1(r=0.441,p<0.001),MMP-9/TIMP-1(r=0.241,p<0.01),respectively. Conclusions Plasma MMP-9,TIMP-1,MMP-9/TIMP-1,and IMT,whole plaque score,soft plaque score and hard plaque score in carotid artery in ACS group are significantly higher than those in SAP group and control group.Plasma MMP-9,TIMP-1,MMP-9/TIMP-1 have positive relationship with IMT,whole plaque score and soft plaque score in carotid artery,respectively.

Abstract:Aim To explore the stability of different morphologic types of plaque and its predict value in unstable angina. Methods 85 unstable angina patients and 40 control subject underwent coronary angiography.According to the morphologic types of plaque,the patients with unstable angina were divided into type I(smooth borders) group(n=21) and type Ⅱ (irregular lesions) group(n=45) and type Ⅲ(long lesions) group(n=19).The plasma factor Ⅶ clotting activity and the plasma concentrations of tissue type plasminogen activator(tPA),plasminogen activator inhibitor(PAI),fibrinogen(FG) and D-dimer were measured respectively.56 unstable angina patients were follow-up one year. Results The plasma coagulative and fibrinolytic parameters of the unstable angina group was more extraordinary than those of the control group.In unstable angina group,the coagulative activity of type Ⅱ group was higher than type I and type Ⅲ group(p<0.05) and cardiac events such as acute myocardial infarction and sudden cardiac death were also higher within one year(p<0.01). Conclusion Each morphologic types of plaque were different in both coagulative and fibrinolytic activity and can be used to judge the stability of unstable angina's plaque and to predict prognosis.

Abstract:Aim To investigate the associations between soluble intercellular adhesion molecule-1(sICAM-1) and high-sensitivity C-reactive protein(hs-CRP) with atherosclerosis in type 2 diabetes mellitus patients(T2DM). Methods The levels of sICAM-1 and hs-CRP in T2DM with or without atherosclerosis and normal comtrols was determined,sICAM-1 was measured by enzyme-linked immunosorbent assay,hs-CRP was measured by immuoturbidimetric method.The carotid intima-media thickness(IMT) and carotid plaques were measured by ultrasound. Results The levels of sICAM-1 and hs-CRP in T2DM with or without atherosclerosis were higher than those in normal comtrols(p<0.01),and the levels of sICAM1 in T2DM with atherosclerosis were higher than that in T2DM without atherosclerosis(p<0.01). Conclusions sICAM-1 is a special risk factor for atherosclerosis in T2DM,but hs-CRP is not.

Abstract:Aim To investigate the association between myocardial infaction(MI) and functional matrix metalloproteinase-9 polymorphism(C1562T). Methods A case-control study of seventy-eight patients with angiographically documented MI and eighty-one control subjects with a normal angiogram was conducted.Genotype was determined by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) for the common C1562T functional promoter polymorphism of the MMP-9 gene. Results The results of individual polymorphisms analysis showed that the frequency of C/T genotype and 1562T allele of MMP-9 gene of MI patients(26.9% and 13.5%) were significantly higher than that in control group(9.9% and 4.9%;p<0.01). Conclusion The present findings suggest that the genetic polymorphism in MMP-9 promoter(C1562T) is associated with the susceptibility to MI in the Han population of China.

Abstract:Aim To study the relationship between endothelium-dependent vasodilatation in abdominally obese patients and serum leptin. Methods The patients were matched into abdominally obese group and controll group according to age and sex.The concentration of leptin was assayed by radioimmunoassay.The endothelium-dependent vasodilatation was measured by high-resolution ultrasound in the two groups. Results The brachial flow-mediated vasodilatation in abdominally obese group was much lower compared with that in control group(6.71%±3.60% vs 13.81%±3.71%,p<0.01).However,there was no significant difference in response to nitroglycerin between the two groups(19.71%±6.63% vs 18.60%±6.35%,p>0.05).The concentration of serum leptin in abdominally obese group was much higher than that in control group,the obese male vs the controlled male is 8.63±3.73 μg/L vs 3.05±1.56 μg/L(p<0.01);the obese female vs the controlled female is 16.73±6.93 μg/L vs 7.93±3.66 μg/L(p<0.01).The brachial flow-mediated vasodilatation was negatively correlated with the concentration of serum leptin according to Pearson correlation analysis. Conclusions The endothelium-dependent vasodilatation dysfunction in abdominally obese patients is negatively correlated with the increase of the concentration of serum leptin.

Abstract:Aim To investigate the relationship between carotid atherosclerosis,C-reactive protein(CRP) and acute cerebral infarction. Methods Histological changes of carotid artery were observed by color Doppler and the serum CRP were evaluated in 86 patients with acute cerebral infarction,compared with the control group which contained 40 non-cerebral infarction patients. Results Incidence of carotid artery endothelial plaque in patients with acute cerebral infarction was significantly higher than those of control group(p<0.01). In patients with acute cerebral infarction,soft plaque and mixed plaque were predominant while hard plaque were predominant in control group,there were significant difference between the two groups(p<(0.01)).Serum CRP levels in patients with cerebral infarction were obviously higher than those of control group(p<0.01).Serum CRP levels in 5 groups were respectively 9.80±2.43 mg/L,10.72±2.55 mg/L,7.46±2.54 mg/L,6.38±1.96 mg/L and 6.15±1.71 mg/L,which histological changes of carotid artery endothelial were soft plaque,mixed plaque,hard plaque,crude endothelium and normal,the levels of serum CRP in soft plaque group and mixed plaque group were higher than those of hard plaque group,crude endothelium group and nomal group(p<0.01). Conclusion Incidence of carotid artery endothelial plaque and serum CRP levels were obviously increased in patients with acute cerebral infarction.Increase of the serum CRP levels can represent the properties and stability of carotid artery plaque.

Abstract:Aim To investigate the relationship between apolipoprotein A5 gene-1131T>C polymorphism,S19W polymorphism and the serum lipid levels in Chinese patients with endogenous hypertriglyceridemia. Methods The restriction fragment length polymorphism(RFLP) of-1131T>C and S19W was studied using polymerase chain reaction(PCR) in 200 healthy subjects and 182 subjects with endogenous hypertriglyceridemia from a population of Chinese Han nationality in Jiangsu Province. Results The frequencies of T,C alleles of-1131T>C polymorphism were 52.7%,47.3% and 67.0%,33.0% in HTG group and control group,respectively. There were significant differences in allele frequencies between HTG group and control group(p<0.05). The relationship between S19W polymorphism and the disease risk was not found. Conclusion C allele of apolipoprotein A5-1131T>C polymorphism is associated with higher serum TG.