Abstract:Aim To investigate the effects of Stichopus Variegates on proliferation and apoptosis induced by platelet-derived growth factor-BB(PDGF-BB)in rat vascular smooth muscle cells(VSMC),and to explore its mechanism of anti-atherosclerosis role,prevention from restenosis after percutaneous transluminal coronary angioplasty(PTCA).Methods The tissue plate method was employed to culture rat thoracic aorta smooth muscle cells in vitro.The effects of Stichopus Variegates on proliferation were analysed using methyl thiazolyl tetrazolium(MTT)colorimetry and flowcytomerty.Cell apoptosis was investigated by flowcytomerty and TUNEL methods.Results Treatment of VSMC with PDGF-BB led to promote proliferation and suppress apoptosis,oppositely Stichopus Variegates significantly reversed these effects.MTT colorimetry showed mean OD value decreased from 0.406±0.003 to 0.187±0.017(p<0.01).Flow cytometry showed the percentage of G0/G1 phase cells increased from 77.4%±5.7% to 88.1%±5.3%(p<0.05)and the percentage of apopototic cells increased from 8.6%±2.3% to 22.6%±2.3%(p<0.05).TUNEL results showed that the percentage of apopototic cells increased from 2.7%±0.4% to 10.1%±0.9%(p<0.01).Conclusion Stichopus Variegates significantly inhibited PDGF-induced proliferation and promoted cell apoptosis of VSMC,which may play important roles in the prevention of atherosclerosis and restenosis after PTCA in vivo.

Abstract:Aim To investigate the involvement of stromal cell-derived factor-1(SDF-1)/CXCR4 Axis in recruitment of endothelial progenitor cell(EPC)and reendotheliazation after vascular injury.Methods EPC migration induced by SDF-1α was determined with modified Boyden chamber assay.Treatment of mice after carotid injury with EPC or EPC coincubated with AMD3100(an antagonist of CXCR4),recruitment of EPC,reendotheliazation and neointimal lesion area were determined 14 d later.Results SDF-1α profoundly enhanced EPC migration(p<0.01),but could not enhance migration of EPC treated with AMD3100(p>0.05).Transfused EPC were strictly restricted to the injury site(14.2±3.6 cell/section),and lectin binding confirmed the endothelial phenotype,but only few EPC coincubated with AMD3100 were recruited to the injury site(4.0±2.5 cells/section).Treatment with EPC caused enhanced reendothelialization(83.45%±5.44%,p<0.01)associated with a reduction of neointima formation(19 237±1 875 μm2,p<0.01)compared with isotype control(66.46%±6.16%,34 676±2 412 μm2);treatment with EPC coincubated with AMD3100 caused no enhanced reendothelialization(68.02%±6.68%,p>0.05)and no reduction of neointima formation(32 451±2 081 μm2,p>0.05)compared with isotype control.Conclusions Stromal cell-derived factor-1/CXCR4 Axis plays an instrumental role in recruitment of EPC and reendotheliazation after vascular injury in mice.

Abstract:Aim To investigate the effects of the Lobelia Chinensis Lour Alkaloids(LCLA)on endothelin-1(ET-1)mRNA expression and endothelin synthesis and release in renal hypertensive rats(RHR).Methods The glodblatt renovascular hypertension model was induced in rats by two-kidney one clip method.After 4 weeks the rats were divided into 3 groups,including the hypertensive group,the captopril group and the LCLA group,while the control group was given sham-operation.Using in situ hybridization to observe ET-1 mRNA expression in peripheral blood leucocytes.Using immunohistochemical technique to count positive rate of ET in “en face” preparations of arterial endothelium to reflect the synthesis of ET.Using radioimmunoassay to examine the level of ET in serum.Results Compared with the sham operation group,the positive rate of ET-1 mRNA in peripheral blood leucocytes(34.64%±8.39% vs 9.34%±4.47%,p<0.05),the positive rate of ET in arterial endothelium(7.42%±0.24% vs 1.58%±0.24%,p<0.05)and the plasma concentration of ET(221±24 ng/L vs 138±19 ng/L,p<0.05)were increased significantly in hypertensive rats.After treating with LCLA for 8 w,compared with the hypertensive group,ET-1 mRNA expression(20.38%±11.31% vs 34.64%±8.39%,p<0.05),endothelin(ET)synthesis(3.53%±0.21% vs 7.42%±0.24%,p<0.05)and release(191±21 ng/L vs 221±24 ng/L,p<0.05)were significantly inhibited.Conclusions The expression of ET was increased in RHR.The LCLA can inhibit the expression of ET in the transcriptional and translational level which may be effective to the preventing and treatment of the renal hypertension.

Abstract:Aim To explore the effect of stromal cell derived factor 1α—CXCR4(SDF-1α—CXCR4)on THP-1 cells chemotaxis,the influence of oxidized low density lipoprotein(ox-LDL)on THP-1 cells migration exposed to SDF-1α,and the effect of ox-LDL on the expression of CXCR4 in THP-1 cell.Methods THP-1 cells chemotaxis was examined with transwell permeable supports.CXCR4 mRNA and protein was revealed by RT-PCR and Western blot respectively in THP-1 cells incubated with different concentrations/time of ox-LDL.Results SDF-1α induced a pronounced migration of input cells in a concentration-related manner,but this effect was obviously inhibited by the antibody to CXCR4.After pretreated with different concentration of ox-LDL for 48 h,THP-1 cells were exposed to 10 μg/L SDF-1α,ox-LDL enhanced THP-1 cells migration in a concentration-dependent manner and 50 mg/L ox-LDL support 11 fold higher of cells migration.CXCR4 was constitutionally expressed in THP-1 cells.Compared with the control group,a four-to-five fold induction of CXCR4 occurred in THP-1 treated with 50 mg/L ox-LDL.The up-regulation effect was monitored within 6 h,it was peaked at 12 h,then followed with a decline,which mirrored the expression pattern of CXCR4 in monocyte.Conclusions SDF-1α/CXCR4 is implicated in chemotaxis of THP-1 cell and chemotactic response of monocyte to SDF-1α is enhanced by ox-LDL;ox-LDL up-regulates CXCR4 expression.

Abstract:Aim To investigate interleukin-10(IL-10)affecting chemokine mRNA expression in monocyte-macrophage derived foam cells and its mechanism.Methods Macrophage was induced by phogrbol myristate acetate(PMA)forming THP-1 cell,and then the cells were further stimulated by oxidized low density lipoprotein with or without IL-10.The influence by IL-10 on chemokine mRNA expression was observed by reverse transcription-polymerase chain reaction(RT-PCR)and the nuclear factor-κB(NF-κB)activation was observed by electrophoretic mobility shift assay(EMSA).Results IL-10 could selectively inhibit monocyte chemoattractant protein-1(MCP-1)and macrophage inflammatory protein-5(MIP-5)mRNA expression.The effects were in dose-dependent fashion.However there was no effect on IL-8 mRNA expression.IL-10 could also significantly inhibit ox-LDL induced NF-κB activation.Conclusions The inhibition on NF-κB activation induced by IL-10 is responsible for decreasing MCP-1 and MIP-5 mRNA expression.The reduction of chemokine expression at the site of atherosclerotic plaque might cut down inflammatory cells recruitment and reduce the plaque further formation.

Abstract:Aim To compare the different effects of perindopril and losartan on kidney,and investigate the mechanisms of protective roles of these two drugs.Methods The investigation comprised normotensive control,hypertensive group,perindopril group and losartan group,of which blood pressure,nitric oxide(NO)level,microalbuminuria,renal function and relevant ultrastructural indexes in vessel and glomerulus with transmission electronic microscope were acquired.Results In three-month old hypertensive rats,electronic microscopic observation demonstrated early impairment of vessel endothelium and slight lesion of glomerular basement membrane compared with age-matched controls.Also,the hypertensive rats had lower level of renal NO and elevation of microalbuminuria.At eight-month old,aggravation of impairment of vessel endothelium and lesion of glomerulus has been shown.Microalbuminuria was associated with lesion of endothelium and glomerulus and renal NO level,but not related to blood pressure.With the treatment of perindopril or losartan,not only reduction of microalbuminuria was achieved but also elevation of NO and decrease in volume density of vessel endothelium,mesangial cells and mesangial area were shown.In addition,NO level was higher in perindopril group than that of losartan.Conclusions Endothelial dysfunction seems to be the most plausible cause of microalbuminuria in hypertension.By way of blocking Ang Ⅱ and improving endothelial function,perindopril and losartan can inhibit the deteriorations of kidney more than blood pressure lowering.Moreover,perindopril has more significant effect on elevating NO level.

Abstract:Aim To observe the peroxisome proliferator-activated receptor-γ coactivator-1(PGC-1)expression in rat skeletal muscle after the low-intensity swimming exercise,and to determine whether the PGC-1 is involved in the regulation of rat energy metabolism.Methods 26 Wistar rats were divided into 3 groups:9 in control group,9 in high-fat feeding group and 8 in swimming exercise following high-fat feeding group(exercise group).PGC-1 mRNA expressions were determined by reverse transcription polymerase chain reaction(RT-PCR).Results Compared with control group,serum tryglyceride(TG),total cholesterol(TC)and low density lipoprotein cholesterol(LDLC)levels were elevated in high-fat feeding group(p<0.05,p<0.01);Compared with high-fat feeding group,serum TG,TC and LDLC levels were decreased(p<0.05),while high-density lipoprotein cholesterol(HDLC)was increased in exercise group(p<0.05).Low-intensity of swimming induced a significantly elevated expression of PGC-1 mRNA in rat skeletal muscle,compared with the control rats(p<0.01)and high-fat feeding rats(p<0.05).PGC-1 protein was also elevated in swimming rats than those in control and high-fat feeding rats(p<0.05).Conclusion Exercise training improved the energy metabolisms of skeletal muscles via PGC-1 in rats,and thus may altered the hyperlipidemia as well as the course of atherosclerosis.

Abstract:Aim To investigate the effects of early estrogen replacement therapy(ERT)of different doses on atherosclerotic lesions,serum lipid and monocyte chemoattractant protein-1(MCP-1)levels in ovariectomized fat-fed female rabbits.Methods Twenty-eight female New Zealand White rabbits were randomly divided into sham operation group,ovariectomized group,ovariectomized with low-dose ERT group(estradiol benzoate 200 μg)and ovariectomized with high-dose ERT(estradiol benzoate 1 mg)group.All rabbits were given high-fat diets after the surgery for 12 weeks.The levels of serum total cholesterol(TC),low density lipoprotein cholesterol(LDLC),high density lipoprotein cholesterol(HDLC),estradiol and MCP-1 were measured.12 weeks later,the aortas were taken for pathological analysis and for calculating the areas of atherosclerotic plaques.Results Compared with sham operation group,the serums levels of TC,LDLC and MCP-1 in ovariectomized group were significantly increased after high-fat diet for 12 weeks,meanwhile the serum levels of HDLC and estradiol were decreased.But the above indexs had no differences between ERT groups and sham operation group.Compared with low-dose ERT group,high-dose ERT group had higher levels of estradiol and lower levels of MCP-1.After high-fat diet for 12 weeks,the extent of atherosclerotic lesions of aorta in ovariectomized group was remarkably bigger than that in sham operation group.The extent in both ERT groups was significantly smaller than that in sham operation group and ovariectomized group.There were no statistical difference in the aortic lesions among two ERT groups.The areas of atherosclerotic plaque were positively correlated with the serum levels of LDLC,TC,and MCP-1,but negatively correlated with the serum levels of HDLC and estradiol.Conclusions Early ERT can modulate the blood lipid,decrease the serum levels of MCP-1 and reduce the areas of atherosclerotic plaque.The antiatherogenic effect of early ERT may be associated with its effects of ameliorating the lipid metabolism and anti-inflammation.

Abstract:Aim To investigate the effects of rosiglitazone on the expression of angiotensin Ⅱ(AngⅡ)type 1 receptor(AT1R)and AngⅡ type 2 receptor(AT2R)in AngⅡ-induced vascular smooth muscle cells(VSMC)and possible molecular mechanisms of anti-atherosclerosis.Methods VSMC of mice were cultured by the explant-attachment method.Using semi-quantitative reverse transcription polymerase-chain reaction(RT-PCR)and immunochemistry,we measured the dose and time dependent effects of rosiglitazone on AT1R and AT2R mRNA and protein in AngⅡ-induced mice VSMC.Results Slight expressions of AT1R and AT2R were observed in cultured VSMC in vitro.AngⅡmarkedly upregulated the expression of AT1R mRNA and protein(p<0.01).Meanwhile,AngⅡdownregulated the expression of AT2R mRNA and protein(p<0.01 and 0.05).Twelve hours post different concentration rosiglitazone(20,30,50 μmol/L)treatment,the expression of AT1R mRNA and protein of VSMC were significantly attenuated(p<0.01,vs AngⅡ group).However,the expression of AT2R mRNA and protein of VSMC were markedly upregulated(p<0.01,vs AngⅡ group).At 6 hours post 30 μmol/L rosiglitazone treatment,the expression of AT1R mRNA and protein decreased,and then the expression of AT2R mRNA and protein increased,reaching a maximum at 24 h post treatment(p<0.01).Conclusions Rosiglitazone can not only downregulate the expression of AT1R mRNA and protein,but also upregulate the expression of AT2R mRNA and protein in AngⅡ-induced VSMC in concentration-dependent and time-dependent manners.Above effects may be crucial mechanisms of the anti-inflammation and anti-atherosclerosis of peroxisome proliferator-activated receptor-γ agonists.

Abstract:Aim To investigate if the atorvastatin calcium protects the brain against injury induced by ischemia/reperfusion(I/R)and its impact on the expression of nuclear factor-κBp56(NF-κBp56)and intercellular adhesion molecule-1(ICAM-1).Methods Rats were subjected to cerebral I/R injury through the occlusion of the middle cerebral artery.Neurological deficits were determined by Longa's score.The expression of ICAM-1 and NF-κB were detected by immuno-histochemical analysis.The effusion of polymorphonuclear leucocytes(PMNLs)in brain tissue was evaluated by HE staining.Results In group I/R+statin,contrasting to group I/R,the expression of ICAM-1 and NF-κB were decreased significantly(p<0.05);the Longa's score were lower(p<0.05);and the effusion of PMNLs were inhibited(p<0.05).Conclusion Atorvastatin-calcium has protective effects on the brain after experimental stroke and its probable mechanism involves the decreased expression of NF-κBp56 and ICAM-1.

Abstract:Aim To investigate the effects of advanced glycation end product(AGE)on cellular proliferation and expression of plasminogen activator inhibitor-1(PAI-1)in cultured rat aortic vascular smooth muscle cell(VSMC).Methods Primary cultures of smooth muscle cell from rat aorta were exposed to AGE of different times and different concentrations.3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT)assay was adopted for the quantification of the cell proliferation ratio and PAI-1 expression was determined by reverse-transcription polymerase chain reaction(RT-PCR).Results Cell proliferation in different AGE concentration medium was greater than that in nonglycated BSA medium(p<0.05),except for 8 hours.After intervention of AGE,plasminogen activator inhibitor-1 mRNA was increased markedly(p<0.001)in a time-related manner.The expression of plasminogen activator inhibitor-1 mRNA was upregulated in a dose-related manner(0.85±0.05,0.97±0.05,1.08±0.12,1.41±0.05),in comparison with that in control group(0.80±0.03,p<0.05).Conclusion AGE induced proliferation and the expression of PAI-1 mRNA in VSMC.This may be involved in the pathogenesis of atherosclerosis in patients with diabetes mellitus.

Abstract:Aim To investigate the changes of matrix metalloproteinase-2 and 9(MMP-2,MMP-9)in rat cardiac hypertrophy,and the effects of Pentoxifylline(PTX)on it in a rat model.Methods Twenty-four male SD rats were randomly divided into control group,norepinephrine(model)group and norepinephrine + PTX(treated)group.The rat cardiac hypertrophy models were established by intraperitoneal injection of norepinephrine(NE)twice a day for 15 days.Extracellular matrix remodeling was evaluated by morphological examination,stained with Van-Gieson(VG).The content of hydroxyproline in the tissue of myocardium was measured.The protein expression of MMP-2 and MMP-9 were examined by immunohistochemical analysis.Results NE-induced hypertrophy and extracellular matrix remodeling predominantly occurred in the left ventricular,the expression of the collagen(1.929±0.514 mg/g vs 1.009±0.442 mg/g,p<0.01)elevated obviously;the average of the grave scale of protein expression of the MMP-2(131.1±9.8)and MMP-9(125.3±4.1)were lower than those of the control group obviously(p<0.01);After PTX treatment,the collagen(1.151±0.215 mg/g)was decreased and the average of the grave scale of protein expression of MMP-2(153.5±6.9)and MMP-9(149.5±5.3)were higher than those of the model group prominently(p<0.01).Conclusion Pentoxifylline can prevent the cardiac hypertrophy and extracellular matrix remodeling,which was associated with the attenuation of myocardial MMP-2 and MMP-9.

Abstract:Aim To investigate the effect of angiotensin Ⅱ(AngⅡ)on the expression of monocyte chemoattractant protein-1(MCP-1)receptor CCR2 and therapeutic effect of losartan on atherosclerosis.Methods Human monocytoid cell(THP-1)were treated with AngⅡ(10-7 mol/L)for 24 h in the absence or presence of losartan(10-7,10-6,10-5 mol/L).The level of MCP-1 in the medium,the adhesion of monocytes to endothelial cell and the expression of CCR2 mRNA were determined.Results Compared with control,incubation of THP-1 with AngⅡ(10-7 mol/L)for 24 h significantly elevated the concentrations of MCP-1(26.46 ±3.58 ng/L vs.10.56 ±2.34 ng/L,p<0.01),increased the number of monocytes binding to endothelial cell(596±27 vs.268±16,p<0.01)and upregulated the expression of CCR2 mRNA.Treatment with losartan decreased the levels of MCP-1,inhibited monocytic binding to endothelial cell,and downregulated the expression of CCR2 mRNA by AngⅡ.Conclusions These data suggest that treatment with losartan in vitro inhibits monocytic activation via downregulation of MCP-1 receptor CCR2 genetic expression.

Abstract:Aim To observe the levels of nitric oxide,endothelin-1 and soluble P-selectin in patients with obstructive sleep apnea/hyponea syndrome and determine whether continuous positive airway pressure treatment has effect on them.Methods Blood concentration of nitric oxide,endothelin-1 and soluble P-selectin,the markers of vascular endothelial cell function,were determined by nitrate reductase,radioimmunoassay,and enzyme-linked immunoassay,respectively.Results Compared with healthy controls(83.26±47.74 μmol/L,58.49±15.02 ng/L and 32.90±55.73 mg/L;respectively),the levels of nitric oxide,endothelin-1 and soluble P-selectin both in patients without cardiovascular disease(63.41±27.68 μmol/L,71.56±23.33 ng/L and 131.13±77.93 mg/L;p<0.05)and in those with cardiovascular disease(59.59±34.19 μmol/L,89.01±44.87 ng/L and 160.83±125.45 mg/L;p<0.05)were significantly changed.However,after continuous positive airway pressure treatment,the blood concentration of nitric oxide(60.46±23.53 μmol/L vs 75.11±23.91 μmol/L;P=0.025),endothelin-1(73.60±20.78 ng/L vs 61.84±18.79 ng/L;P=0.031)and soluble P-selectin(137.34±90.35 mg/L vs 89.56±34.60 mg/L;P=0.013)were significantly improved.Conclusions Obstructive sleep apnea/hyponea syndrome patients,no matter whether complicated by cardiovascular diseases or not,have significant vascular endothelial cell dysfunction,which can be improved through the continuous positive airway pressure treatment.

Abstract:Aim To study the protective effects of luomaishutong granule on myocardium after reperfusion in patients with acute myocardium infarction(AMI).Methods The research was performed on the patients with AMI whose initial ECG showed ST segment elevation and the patients who received percutaneous coronary intervention(PCI)immediately after onset.All patients were classified randomly into two groups:control group in which the patients were given routine drug treatment(50 cases)and treatment group in which the patients were given luomaishutong granule(50 cases).The course of treatment is two weeks.We examined the blood interleukin 6(IL-6)mRNA and high-sensitivity C-reactive protein(hs-CRP)levels on the 1st day,7st day after percutaneous coronary intervention.We observed the left ventricular ejection fraction(LVEF)in two dimensional echocardiography at the end of the fist week and the second week.At the same time we also observed the incidence rate of arrhythmogenesis and degree of ST segment resolution.Results The day after the PCI IL-6 mRNA and hs-CRP in the patients of the two groups increased and there is no difference between the two groups.But seven days later,IL-6 mRNA and hs-CRP were decreased much more than before(p<0.01).Compared with control group,in treatment group IL-6 mRNA and hs-CRP were also decreased(p<0.05),and the incidence rate of arrhythmogenesis and degree of ST segment resolution between the two groups had obvious difference(p<0.05),and LVEF was improved obviously(p<0.05).Conclusions Luomaishutong granule can prevent myocardial ischemia-reperfusion injury,improve myocardial reperfusion,restrain inflammatory reaction and improve LVEF.

Abstract:Aim To explore the prognostic value of measurements of Troponin T(TnT),high sensitive C-reactive protein(hs-CRP),and B-type natriuretic peptide(BNP)in patients with non-ST elevation acute coronary syndrome(ACS).Methods Levels of serum TnT,hs-CRP,and BNP were measured among 145 ACS patients verified by coronary angiography.Patients with ACS were followed up 12 months.The end point was cardiac events including new or recurrent myocardial infarction(MI)and cardiac death.Results The multivariate Logistic regression mode analysis revealed that TnT,hs-CRP,and BNP were independent long-term prognostic markers in patients with non-ST elevation ACS.The number of elevated biomarkers remained a significant predictor of the composite endpoint after adjustment for known clinical predictors.Conclusions Measurements of TnT,hs-CRP,and BNP had practical significance in evaluating the long-term outcome of non-ST elevation ACS.

Abstract:Aim To investigate the effect of obesity on the arteriosclerosis development in children by comparing the risk factors for arteriosclerosis in obese children with those in non-obese children.Methods Fifty-one obese children and thirty-two non-obese children were evaluated for endothelium-dependent arterial dilation,intima-media thickness(IMT)of the common carotid artery,the maximum preperitioneal fat thickness(Pmax)and minimum subcutaneous fat thickness(Smin)by B ultra sonograthy;24 h average systolic blood pressure(SBP)and diastolic blood pressure(DBP)were detected;the serum levels of triglyceride(TG),total cholesterol(TC),blood glucose and free fatty acids(FFA),low density lipoprotein cholesterol(LDLC),high density lipoprotein cholesterol(HDLC)were deteced by chemistry;insulin(FIns),by RIA;adiponectin(AD),resistin by ELISA,and high-sensitivity c-reactive protein(hs-CRP),by scatter nephelomete method.Results The values of Hs-CRP,HOMA-IR,INS,Smin,Pmax,SBP,TG,LDLC,and FFA were higher in the obese children than in the non-obese children(p<0.05),while HDL-C and adiponectin in the obese children were lower than those in the non-obese children(p<0.05).However,no statistical difference was found between the levels of IMT,DBP,and Endothelium-dependent arterial dilation in the obese children and those in the non-obese children(p>0.05).The BMI was correlated with SBP(r=0.43,P=0.006),hs-CRP(r=0.461,P=0.018),HOMA-IR(r=0.463,P=0.007),FIns(r=0.404,P=0.013),FFA(r=0.358,P=0.018),while it was negatively correlated with AD(r=-0.356,P=0.031).However,there was no statistically different relationship between ITM and other biochemical indicators.Conclusions The levels of the risk factors for arteriosclerosis were much higher in obese children than those in non-obese children.The results showed that the chronic inflammation situation of arteriosclerosis has already existed in the obese children,but no morphological change in the endarterium was found.