Abstract:Aim To determine whether advanced glycation end products(AGE)altered the morphology and distribution of the adherens junction protein vascular endothelial(VE)-cadherin in human umbilical vein endothelial cells(hUVEC)and to understand the mechanism of this alteration.Methods Human umbilical vein endothelial cells(hUVEC)were respectively incubated with AGE-HSA in different concentrations and timing.In some other cases,hUVECs were pretreated with mitogen activated protein kinase(MAPK)pathway inhibitors(PD98059,SP600125,SB203580),or transfected with adenovirus.To visualize the morphological changes of VE-cadherin,cells were incubated with rabbit anti-VE-cadherin primary antibody and then FITC anti-rabbit IgG secondary antibody.The morphological changes of VE-cadherin were observed with confocal microscope.Results The morphological of VE-cadherin was significantly changed in coincident with an increase of the dose and time of AGE-HSA.These changes could be inhibited with MAPK and Rho inhibitors.Conclusion These observations suggested that AGE modified proteins can induce morphological changes of VE-cadherin in endothelial cell.Activations of MAP kinase and Rho kinase pathways play an important role in AGEs-induced hUVECs VE-cadherin distribution.

Abstract:Aim study the change of expression of matrix metalloproteinase-9(MMP-9)and the permeability of blood brain barrier(BBB)in the rats model with cerebral ischemic and reperfusion.Methods A right middle cerebral artery ischemia-reperfusion models were established in this experiment,the change of the permeability of BBB had been evaluated by mensurating the content of Evan's blue(EB),The expression of MMP-9 in ischemia reperfusion rat brains had been investigated by immunohistochemistry technology.Result After cerebral ischemia-reperfusion,the mount of EB exudation of the brain tissues begin increase in 3 h(0.18±0.08 vs 0.86±0.12 μg/g),and will get its peak at the time of 1 d(4.64±1.27 μg/g),and will decrease gradually 2 d later(4.48±0.65 μg/g).The expression of MMP-9 increases notablely at 12 h(3.64±0.56 vs 6.76±2.26),1 d(11.24±2.64),2 d(21.52±2.03),4 d(16.66±1.38),and 7 d(14.64±3.77)later after ischemia-reperfusion,and will go to its peak at the second day.Conclusion The expression of MMP-9 positive cells presents time-related with the increase of BBB permeability after cerebral ischemia-reperfusion,which point out the MMP-9 participated the process of the damage of BBB.

Abstract:Aim To investigate the effects of probucol on high fat-high cholesterol(HFHC)diet-induced hyperlipidemia and oxidative stress in apo E deficient(apoE-/-)mice and genetic background C57BL/6J mice.Methods The hyperlipidemic mice were induced by feeding HFHC diet which was supplemented with 15%lard and 0.25% cholesterol.The levels of plasma lipids were determined by standard assays.The oxidative stress was evaluated by the serum levels of total antioxidant capacity(T-AOC),superoxide dismutase(SOD),and malonic dialdehyde(MDA)determined by commercially provided kits.The aortic atherosclerotic lesions were demonstrated by Sudan IV staining.Activity of paraoxonase 1(PON1)was assessed by modified Echerson's method.Expression level of PON1 in liver was evaluated by semi-quantitative reverse-transcription-polymerase reaction(RT-PCR).Results HFHC diet elevated the levels of serum triglyceride(TG),total cholesterol(TC),low density lipoprotein-cholesterol(LDLC),and high density lipoprotein-cholesterol(HDLC).Probucol decreased the elevation of serum levels of lipids.In C57BL/6J mice,the levels of TG,TC,LDLC,and HDL-C lowered 41%,43%,44.8%,and 41.3%,respectively,compared with control group(P<0.05).In apo E-/-mice,decrease of 36.2%,28.2%,31.3%,and 33.9%,respectively,was observed versus apo E-/-control(P<0.05).Probucol treatment prevented the atherosclerotic lesion development in both C57BL/6J and apoE-/-mice.Compared with HFHC groups,the aorta lesions were significantly decreased by 61.1% in C57BL/6J mice and by 27.2% in apo E-/-mice(P<0.05),respectively,after treatment of probucol.Admi-nistration of probucol lowered the serum levels of MDA and oxLDL,and elevated the levels of serum T-AOC and SOD.The results also demonstrated a more significant increase of PON1 activity and expression level in C57BL/6J mice than in apo E-/-mice.Conclusion Probucol decreases the levels of serum lipids and alleviates the atherosclerotic lesions in HFHC diet-induced hyperlipidemic mice.Probucol may contribute to a significant decrease of the oxidative stress.The effects of probucol may be attenuated by apo E deficiency in mice.

Abstract:Aim To explore effects of telmisartan and angiotensin Ⅱ(AngⅡ)on expression of angiotensin converting enzyme-2(ACE-2)in thoracic aortic smooth muscle cells(A10 cells).Methods The expression of ACE-2 protein was tested by Western blotting analysis and the expression of ACE-2 mRNA was determined with reverse transcription-polymerase chain reaction(RT-PCR)technique.Results Telmisartan at the concentration from 10-8 to 10-5 mol/L dose-dependently enhanced the expression of ACE-2 protein;within 0 to 24 h,10-6 mol/L telmisartan increased ACE-2 protein expression in a time-dependent manner.Moreover,telmisartan at 10-6 mol/L upregulated the expression of ACE-2 mRNA.AngⅡ at 10-8 to 10-5 mol/L downregulated the expression of ACE-2 protein in a dose-dependent manner.Telmisartan at 10-6 mol/L obviously blocked the inhibition of 10-6 mol/L AngⅡ on the expression of ACE-2 protein and ACE-2 mRNA respectively.Conclusion Telmisartan not only up-regulates the expression of both ACE-2 protein and ACE-2 mRNA in A10 cells,but also antagonizes the inhibition of AngⅡon the expression of both ACE-2 protein and ACE-2 mRNA.

Abstract:Aim To study the effect of stromal cell derived factor 1 alpha(SDF-1α)on CXCR4 expression in THP-1 monocyte,and the effect of SDF-1α and CXCR4 antibody on monocyte/endothelium adhesion.Methods CXCR4 mRNA and protein expressions were detected by RT-PCR and Western blotting respectively in moncyte pretreated with 100 μg/L of SDF-1α for 0 and 30 min.ECV-304 endothelial cells pretreated with 0,50,100 and 200 μg/L SDF-1α for 30 minutes and sub-incubated with THP-1 cells,with or without CXCR4 antibody(10 mg/L),THP-1/ECV-304 adhesion was observed.Results CXCR4 was constitutionally expressed in THP-1 cells and increased remarkably by 100 μg/L SDF-1α for 30 min.THP-1 and ECV-304 adhesion was enhanced by 50～200 μg/L SDF-1α,but it was also apparently inhibited by the CXCR4 antibody(P<0.05).Conclusions SDF-1α enhances CXCR4 expression in THP-1 cells.SDF-1α enhances monocyte/endothelium adhesion,which is related to up-regulating CXCR4 expression.

Abstract:Aim To study the effect of simvastatin on adiponectin(APN)and nuclear factor-kappa B inhibitor kinase(IKK)mRNA expression in adipose tissue of insulin resistant rats.Methods Insulin resistant rat model was induced by high-fat diet feeding.Twenty-five male Wistar rats were randomly divided into two groups fed with either basic chow(n=10)or high-fat diet(n=15)for 10 weeks,then assessed by euglycemic-hyperinsulinemia clamp technique.And rats in high-fat group were randomly divided into simvastatin intervention group(n=5)and non-simvastatin intervention group(n=5).Rats in simvastatin intervention group were fed with simvastatin 10 mg/(kg·d)in gavage.After 5 weeks intervenient treatment,APN and IKK mRNA expressions were tested with RT-PCR.Results The glucose infusion rate(GIR)in high-fat group decreased significantly compared with basic chow group 0.76±0.28 mg/(kg·min)vs 4.26±0.70 mg/(kg·min),P<0.01.There were no obvious difference on APN mRNA expression of adipose tissue between simvastatin intervention group and non-simvastatin intervention group(0.25±0.12 vs 0.29±0.11,P>0.05);but they decreased significantly compared with basic chow group respectively(1.18±0.12,P<0.05).IKK mRNA expression of adipose tissue in simvastatin intervention group decreased significantly compared with non-simvastatin intervention group(0.15±0.03 vs 1.21±0.03,P<0.05),with no obvious difference from basic chow group(0.15±0.03,P>0.05).Conclusion Simvastatin could not increase APN mRNA expression in adipose tissue of insulin resistant rats induced by high-fat diet feeding.But simvastatin may restore IKK mRNA expression in adipose tissue of insulin resistant rats induced by high-fat diet feeding,which maybe benefit from the anti-inflammatory effect beyond the lipid-lowering effect.

Abstract:Aim To investigate the phenotypic modulation and proliferation changes of vascular smooth muscle cell(VSMC)and vascular adventitia fibroblasts(VAF)in the vein graft restenosis model.To evaluate the effects of the phenotypic modulation and proliferation changes of VSMC and VAF in vascular remodeling.Methods To create the pig autogenous vein grafts restenosis model,specimens were harvested at three time points that was 7th,14th and 30th day.Histomorphometrical and immunohistochemical approach were used to detect the changes of vascular remodeling and the expression of proliferation cell nuclear(PCNA),smooth muscle α-actin(SM-α-actin)and osteopontin(OPN).Results ①At 7th day after operation,the neointima formed and continuously thickened,whose thickness reached maximum at 30th day postoperation.Internal elastic lamina(IELA)gradually decreased after operation,and at 30th day group luminal area reduced minimally(P<0.05).Remodeling index and external elastic lamina(EELA)slightly increased at 7th day group,but reduced gradually after that,which at 14th day and 30th day group decreased distinctly(P<0.05).②According to PCNA stain,the proliferation index of VAF and VSMC increased significantly at 7th day after operation,which reached peak at 14th day,whereas the expression of PCNA reverted toward the baseline at 30th day after operation.③According to SM-α-actin stain,at 7th day after operation,the positive cell area decreased in the medium,but adventitia and neointimal were positive;At 14th day,the positive cell area decreased significantly in the medium,however,the positive area of adventitia and neointimal increased significantly;At 30th day positive expression of the medium recovered gradually,and the positive area of adventitia was less than that at 14th day,but the positive expression in the neointimal has no significant changes compared with 14th day.④OPN stain showed that the positive expression was significant in the medium at 7th day after operation.At 14th day,the positive expression of VSMC was less than that at 7th day in the medium,but that of neointimal increased significantly,which reached peak.At 30th day,there was a less positive part of VSMC in the medium,and the positive expression of VSMC in the neointimal decreased significantly compared with 14th day.OPN was negative in adventitia at different time after operation.Conclusion The phenotypic modulation and proliferation changes of VSMC and VAF are the important factors on vascular remodeling,which takes part in and accelerates the course of vein restenosis.

Abstract:Aim To probe the relationship between plaque stability and expression levels of interleukin-6(IL-6),interleukin-18(IL-18)and tumor necrosis factor-α(TNF-α)in human coronary atherosclerosis.And to investigate the correlation of IL-6,IL-18 and TNF-α.Methods One hundred and four autopsy samples of coronary paraffin sections were selected and divided into unstable plaque group,stable plaque group and normal coronary group by means of SmartScape biology miscroscope analysis system.Plaque area,lipid core area and fibrous cap thickness in the three groups were measured and analyzed.The expression levels of IL-6,IL-18 and TNF-α in plaque of human coronary atherosclerosis were measured by immunohistochemical technique.Results There was no statistically and significant difference between the unstable plaque group and the stable plaque group in the area of coronary atherosclerosis plaque(P>0.05),but the lipid core area and fibrous cap thickness were statistically and significanly different between them(P<0.05).IL-6,IL-18 were not expressed,and TNF-α was little or not expressed in the normal coronary group.The expression levels of IL-6,IL-18 and TNF-α were higher in the unstable plaque group than in the stable plaque group,especially in the shoulder of fibrous cap and lipid core(P<0.01).There was positive correlation among the expression of IL-6,IL-18 and TNF-α.Conclusions The expression levels of IL-6,IL-18 and TNF-α in plague of human coronary atherosclerosis was not only related to plaque stability but also correlated among one another.

Abstract:Aim To investigate the influence of granulocyte-macrophage colony stimulating factor on functional activity of rat endothelial progenitor cells in vitro in order to further provide experimental basis for the mechanism of reendothelialization and prevention and treatment of coronary artery disease.Methods Endothelial progenitor cells were separated from bone marrow and spleen of rat with density gradient centrifugation method.Then growth and differentiation procession of endothelial progenitor cells were observed with inverted phase contrast microscope.Endothelial progenitor cells were identified via dual fluorescent staining of FITC-AC133 and PE-vWF.After cultured for seven days,endothelial progenitor cells were added stimulating factors.Then the influence of granulocyte-macrophage colony stimulating factor on proliferation,transfer and blood vessels formation of endothelial progenitor cells were detected with MTT colorimetry,Transwell chamber and Matrigel.Finally the experiment results were analysed.Results Compared with control,granulocyte-macrophage colony stimulating factor significantly increased OD490,the number of migratory cells and tubes(P<0.001).Conclusions Granulocyte-macrophage colony stimulating factor can significantly facilitate proliferation,migration and blood vessels formation of endothelial progenitor cells in vitro.

Abstract:Aim To establish 2-dimensional gel electrophoresis images and seek differentially expressed plasma proteins of unstable angina pectoris patients with blood stasis syndrome.Methods 2-dimensional gel electrophoresis and Matrix-assisted laser desorption/ionization time of flight mass spectrometry were used to screen the differentially expressed plasma proteins among unstable angina pectoris with blood stasis syndrome patients and health volunteers.Results α1-acid glycoprotein,haptoglobin α1 chain,α1-antitrypsin,haptoglobin β chain and haptoglobin α2 chain were significantly highly expressed in the plasma of unstable angina pectoriswith blood stasis syndrome patients,while apolipoprotein AⅣ,apolipoprotein AⅠ and transthyretin were decreased in the plasma of unstable angina pectoris patients with blood stasis syndrome.Conclusions α1-acid glycoprotein,haptoglobin α1 chain,α1-antitrypsin,haptoglobin β chain,apolipoprotein AⅣ and apolipoprotein AⅠ are differentially expressed in the unstable angina pectoris patients with blood stasis syndrome and control group,and some differentially expressed proteins are correlated with inflammatory reaction or lipid metabolic disorder,and these proteins could provide clues for the study and discovery of new protein targets for antianginal drugs.

Abstract:Aim To investigate the correlation between endogenous inhibitor of nitric oxide synthase(NOS)asymmetric dimethylarginine(ADMA)in patients with coronary artery heart disease and severity of coronary atherosclerosis.Methods A total of 75 patients were enrolled and divided into four groups:stable angina pectoris group(SAP,n=15),unstable angina pectoris group(UAP,n=23),acute myocardial infarction group(AMI,n=20)and control group(n=17).Clinical history was collected and total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDLC),high density lipoprotein cholesterol(HDLC)and ADMA levels were determined.Coronary artery lesion was evaluated by Gensinis coring system.Relevance of ADMA to age,sex proportion,TC,TG,LDLC,HDLC,and coronary artery Gensinis coring was analyzed.Results Compared with the control group(3.81±0.48 μmol/L),the serum ADMA level of the SAP group(4.82±0.50 μmol/L),UAP group(5.52±0.55 μmol/L)and AMI group(6.03±1.13 μmol/L)increased significantly(P<0.01).The serum ADMA level of the UAP group and AMI group were higher than that of the SAP group(P<0.01).The difference of the serum ADMA level between the UAP group and AMI group wasn't statistically significant(P=0.279).Gensinis coring of the control group was significantly different from the other three groups(P<0.01).Gensinis coring of the SAP group(17.79±1.86)was significantly lower than those of the UAP group(38.05±4.46)and AMI group(40.62±3.80)(P<0.01).There was nosignificant difference of Gensinis coring between the UAP group and AMI group(P>0.05).Significant correlation between ADMA level and TC,LDLC levels,and Gensinis coring were found(P<0.01).There was no significant correlation between ADMA level and age,sex proportion,TG and HDLC levels(P>0.05).Conclusions The serum ADMA is positively correlative to the degree of coronary atherosclerosis and the morbid state of coronary artery heart disease.It is important for the diagnosis therapy and estimation the morbid state of coronary artery heart disease to observe the changes of the serum ADMA level.

Abstract:Aim To explore the association of serum lipoprotein(a)and serum bilirubin with carotid atherosclerosis in patients with hypertension.Methods 124 patients with hypertension whose carotid atherosclerosis were detected by ultrasound were divided into increased IMT group,atheroma group,mild stenosis group,moderate stenosis group,and severe stenosis group.The serum lipoprotein(a)and serum bilirubin levels were measured using automatic biochemistry analysis system.Results Those patients with more severe carotid atherosclerosis had higher serum lipoprotein(a)and lower serum total bilirubin and serum indirect bilirubin.The level of lipoprotein(a)among five groups above(156.7±89.2 mg/L,237.0±170.3 mg/L,324.0±126.0 mg/L,331.9±197.5 mg/L and 346.2±124.0 mg/L)was significant statistical difference(P<0.05).So were serum total bilirubin(12.4±2.6 μmol/L,10.7±1.4 μmol/L,9.1±1.0 μmol/L,8.1±0.9 μmol/L and 7.1±0.8 μmol/L)and serum indirect bilirubin(9.1±2.4 μmol/L,7.4±1.5 μmol/L,5.9±1.2 μmol/L,4.9±0.8 μmol/L and 3.9±0.8 μmol/L).While Serum direct bilirubin showed no statistical difference between five groups(P>0.05).Otal and indirect serum bilirubin correlated negatively with lipoprotein(a)(r=-0.332 and-0.331,P<0.01 for all).Conclusion High serum lipoprotein(a)and low serum bilirubin may be closely related to atherosclerosis in patients with hypertension.

Abstract:Aim To investigate the relationship of plasma Apelin with left ventricular hypertrophy and carotid artery disease,and to observe the change of plasma Apelin levels in patients with hypertension.Methods 125 essential hypertensive patients and 31 healthy controls were selected.All patients with hypertension were divided into left ventricular hypertrophy group,intima-media thickeness group,carotid artery plaque group and simple hypertension group according to the measuring results by ultrasonography.The levels of plasma Apelin were determined by enzyme immunoassay.Results The plasma Apelin levels were significantly lower in left ventricular hypertrophy group(616±112 ng/L)than in control group(757±176 ng/L)and simple hypertension group(715±170 ng/L,P<0.01 or 0.05).The plasma Apelin levels were decreased in intima-media thickness group(613±133 ng/L)and carotid artery plaque group(592±146 ng/L)compared with control group and simple hypertension group(P<0.001 or 0.05).There were negative correlation between left ventricular mass index,intima-media thickness,carotid artery plaque index and plasma Apelin(respectively r=-0.253,r=-0.255 and r=-0.246,P<0.05).Conclusion Apelin may participate in the pathophysiological process of myocardial hypertrophy and atherosclerosis.

Abstract:Aim To compare the difference of levels of serum oxidized low density lipoprotein(ox-LDL)and high sensitive C-reactive protein(hs-CRP)among different types of coronary heart disease.Methods Levels of serum ox-LDL and hs-CRP and low density lipoprotein cholesterol(LDLC),total cholesterol(TC)and high density lipoprotein cholesterol(HDLC)were measured among 82 patients with coronary heart disease including 42 acute coronary syndrome(ACS),20 stable angina pectoris(SAP)and 20 old myocardial infarction(OMI)and 22 normal healthy control subjects,then the differencs of levels of all four markers were compared among different types of coronary heart disease.Results Serum ox-LDL levels in the coronary heart disease group(62.5±24.8 μg/L)were significantly higher than that in normal healthy control group(42.3±17.9 μg/L,P<0.05),and serum ox-LDL levels in ACS group(68.9±23.4 μg/L)higher than that in SAP group(41.5±21.3 μg/L)and serum ox-LDL levels in OMI group(50.5±21.6 μg/L,P<0.05);serum hs-CRP levels in coronary heart disease group(15.58±4.32 mg/L)were significantly higher than that in normal healthy control group(6.94±1.93 mg/L,P<0.05),and serum hs-CRP levels in ACS group(19.31±1.43 mg/L)higher than that in SAP group(10.29±1.01 mg/L)and serum hs-CRP levels in OMI group(9.56±1.72 mg/L,P<0.05);serum LDLC levels in coronary heart disease group(3.46±1.11 mmol/L)were significantly higher than that in normal healthy control group(2.87±0.82 mmol/L,P<0.05),and serum LDLC levels in ACS group(3.64±0.85 mmol/L)higher than that in SAP group(3.06±1.23 mmol/L)and serum LDLC levels in OMI group(3.40±1.35 mmol/L,P>0.05).Serum TC/HDLC in the coronary heart disease group(3.46±1.11)were significantly higher than that in normal healthy control group(2.87±0.82).Serum TC/HDLC in different types of patients with coronary heart disease were different(P<0.05).Conclusions The levels of serum ox-LDL and hs-CRP and TC/HDLC have differences among different types of coronary heart disease,and these may be targets of coronary heart disease.

Abstract:Aim To study the relationship of ankle brachial index(ABI),atherosclerosis and conventional risk factors.Methods 174 inpatients were performed coronary angiography,Gensini's coronary angiography score was calculated to quantify the extent of coronary artery atherosclerosis.ABI of all subjects were measured,retinal artery was inspected and risk factors of atherosclerosis were investigated.Results ABI between normal coronary angiography group and anormal coronary angiography group had statistically significant difference(P<0.001).There was an inverse linear trend between ABI and Gensini's coronary angiography score(r=-0.768,P<0.001),the more Gensini's coronary angiography score the lower ABI.ABI between normal retinal artery group and anormal retinal artery group had statistically significant difference(P<0.001),the more severity of retinal artery the lower ABI.Gensini's coronary angiography score between normal retinal artery group and anormal retinal artery group had statistically significant difference(P<0.001),the more severity of retinal artery the more Gensini's coronary angiography score.Logistic regression model analysis showed that risk factors including ages,sex,triglyceride,low density lipoprotein,quantity of smoking,diabetes and genetic factor were all associated with ABI significantly.Conclusions ABI is a handy and dependable predictor of atherosclerosis.ABI are all associated with conventional risk factors including ages,sex,triglyceride,low density lipoprotein,quantity of smoking,diabetes and genetic factor.