Abstract:Aim To approach Zhixinkang possible regulation mechanism for atherosclerosis (As) foam cell, and to investigate the effect of Zhixinkang and Lovastatin on CD36 mRNA in foam cell originated from vascular smooth muscle cell (VSMC). Methods The cultured VSMC were loaded with oxidize low density lipoprotein (ox-LDL) and interfered in Zhixinkang and Lovastatin. The expression of CD36 mRNA were detected with flow cytometer and In Situ hybridization and analyzed with statistics. Results Zhixinkang and Lovastatin could decrease CD36 mRNA expression than that in the isotonic Na chloride group and the blank group. Moreover, CD36 mRNA expression in Zhixinkang group was different from that in Lovastatin group. Conclusion Zhixinkang could decrease CD36 mRNA expression, which was superior to Lovastatin. Zhixinkang had the effect of tonifying qi and promoting blood, neutralizing poison and resolving turbid in VSMC, which was one of mechanism to inhibit the multiplication of VSMC and restrain the formation of foam cell.

Abstract:Aim To study the effect of stromal cell derived factor-1 alpha (SDF-1α) on adherence of ECV304/THP-1. Methods Rat SDF-1α gene and pcDNA3.1 were both cut with EcoRⅠ, dephosphorylated, and connected in connecting buffer. Then, the connecting product was transformed to DH5α by calcium chloride, scanned by ampicillin, and cut by enzyme for the right inserting identification, and sub-selected by G418, SDF-1α expression of ECV304 was measured by reverse transcription polymerase chain reaction (RT-PCR) to get a stabilized SDF-1α expressional cells line. Trans-gene ECV304 was seeded in a six-pore plate. Then, THP-1 was added and incubated for 30 minutes, washed with PBS about 3 times, to remove unattached cells. Cells number was counted under microscope from 5 visual field -up, under, left, right and center. Results Stable SDF-1α expression cells line was obtained which was identified by RT-PCR (about 360 bp), and the adherence ability of trans-gene ECV304 was ten times and more than control, and CXCR4 mono-antibody obviously inhibited this adherence ability (P<0.01). Conclusion Endogenous SDF-1α improves adhesion of ECV304/THP-1.

Abstract:Aim To investigate the adhesion regulation effect of stromal cell-derived factor 1α (SDF-1α) on monocytes and endothelial cells induced by oxidized low density lipoprotein (ox-LDL). Methods SDF-1α mRNA and protein were measured by reverse transcript-polymerase chain reaction (RT-PCR) and Western blot respectively in endothelial cells. Cell counting was used to observe the effect of the monocytes and endothelial cells adhesion treated with ox-LDL or SDF-1α antibody. Results SDF-1α was hardly expressed in normal endothelium, but was up-regulated by ox-LDL in a dose and time dependent manner, It peaked at 25 mg/L ox-LDL for 48 h. The adhered amount of monocytes to endothelial cells were 190±15, 226±23, 280±14, 253±8 respectively responding to 1, 5 and 125 mg/L ox-LDL. And the amount was 104±10 in the control (P<0.05). While endothelial cells pretreated with ox-LDL were incubated with SDF-1α antibody with 0.01, 0.1, 1 μg/L respectively, the adhesion of monocytes was 202±17, 142±6, 115±12. It was lower than that of control group significantly (279±11; P<0.05). Conclusion SDF-1α is implicated in monocytes and endothelium cells adhesion.

Abstract:Aim To study whether stromal cell-derived factor-1α (SDF-1α) have effects on endothelial progenitor cells (EPC) and its preliminary mechanism. Methods Bone marrow derived EPC were aquried by Micropore-method and characterized by immunofluorescence staining. The abilities of migration were detected by the methods of transwell migration after EPC treated with different concentrations of SDF-1α. Results SDF-1α promoted EPC biologic activity of migration in a concentration-dependent manner (P<0.05 or P<0.01). Conclusion SDF-1α can improve migration of EPC.

Abstract:Aim To investigate the effect of pcDNA3.1-SDF-1α plasmid on THP-1 cells chemotaxis. Methods pcDNA3.1-SDF-1α plasmid was transfected into ECV304 cells by lipofectamine and the positive clones were screened by G418. The expression of SDF-1α gene in the transfected ECV304 cells were detected by RT- PCR. Then explored the effect of SDF-1α on THP-1 cells chemotaxis with transwell permeable supports. Results G418-resistant ECV304 cell clones were obtained. pcDNA3.1-SDF-1α plasmid effect on THP-1 cells migration. Conclusion SDF-1α recombinant protein obviously induce migration of THP-1 cells.

Abstract:Aim To explore the effect of oxidized low density lipoprotein (ox-LDL) on stromal cell derived factor -1α (SDF-1α) expression in rat vascular smooth muscle cell (VSMC). Methods and Results Primary rat VSMC was incubated with different concentrations/time of ox-LDL, SDF-1α mRNA and protein was revealed by RT-PCR and Western blot respectively. SDF-1α was constitutionally expressed in rat VSMC and was concentration-dependently up-regulated by ox-LDL within 0～50 mg/L, 5 folds up-regulation was induced by 50 mg/L ox-LDL; SDF-1α expression was peaked at 12 h and then followed with a decline, but maintained at 3 folds level of basal expression. Conclusion Oxdizied LDL powerfully induces upregulation of SDF-1α in rat vascular smooth muscle cell.

Abstract:Aim To examine the effect of timisartan on expression and activity in vascular remodeling after artery injury in rats. Methods The rat models of balloon endothelial denudation were builded. Male Wistar rats were randomized to three groups: nomal control group (n=12), model group (n=12) and timisartan group (n=12). The rats in timisartan group were supple diet with timisartan 40 mg/(kg·d). The animals were sacrificed at 30 days after the injury. Western blot were used to detect the expression and activity of focal adhesion kinase (FAK) with RT-PCR or Western blot method. Hyperplasia of intima were observed. Results Compared with the control group, hyperplasia of intima were significantly increased, the expression of FAK mRNA, protein and its activity were significantly increased in model group at 30 days after the injury. Compared with the model group, hyperplasia of intima were significantly lower, the expression of FAK mRNA, the expression of FAK protein and activity were significantly decreased inTimisartan group. Conclusion Timisartan significantly inhibited artery wall remodelling and expression and activity of FAK in the rat model of artery injury. The mechanisms of Timisartan inhibiting artery wall remodelling could involve inhibition of expression and activity of FAK.

Abstract:Aim To evaluate the clinical effects of amlodipine on arterial elasticity and left ventricular hypertrophy in elderly hypertensive patients. Methods The arterial elasticity indexes of the hypertensive patients(76 cases) were detected with CVP rofilor DO-2020,and left ventricular mass indexes were measured with ultrasound cardiography before and after long-term (36 months) treatment with amlodipine. Results the blood pressure was controlled to therapeutic aim and maintained stable after 8 weeks treatment with amlodipine. The small artery elasticity index rised significantly than that after 36 months treatment with amlodipine (2.60±0.77 mL/Pa vs 3.80±2.09 mL/Pa, P<0.01); and the left ventricular mass indexes reduced dramatically than that after therapy (147.3±26.8 g/m2 vs 131.7±24.3 g/m2,P<0.01). Conclusion Amlodipine can improve arterial eladticity and vascular function,retard or even reverse left ventricular hypertrophy,while lowering blood pressure stably.

Abstract:Aim To investigate the effects of Amaryl on newly diagnosed type2 diabetes. Methods 33 newly diagnosed type 2 diabetic patients with fasting plasma glucose (FPG)<12 mmol/L were treated with Amaryl for 12 weeks. Oral glucose tolerance test (OGTT) was performed before and after Amaryl therapy. The fasting and post 2 h plasma glucose (P2hPG), hemoglobin A1C (GHbA1C), Homa-IR, TC, TG, LDLC, HDLC, tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) were compared before and after Amaryl. Results After 2 weeks′Amaryl treatment, good glycemic control was achieved. The FPG, P2hPG, GHbA1C, TC, TG, LDLC, Homa-IR, tPA were lower than before.But HDLC, PAI-1 were higher than before. Conclusion Excellent glycemic controls, improvements of lipid profiles and PAI-1 and decreased insulin resistence can be induced by Amary therapy in newly diagnosed type 2 diabetic patients.

Abstract:Aim To evaluate the relationship between basic fibroblast growth factor (bFGF), carotid artery atheroscleorsis and cerebral ischemicin patients of essential hypertension. Methods B-mode ultrasound examination of common carotid artery (CCA) and internal carotid artery (ICA), bifurcation of carotid artery (BIF) was used. bFGF was measured by enzyme linked immunosorbent assay (ELISA) methods. Results The levels of bFGF (364.5±91.0 mg/L vs 283.4±91.2 mg/L), the incidents of soft plaques (92.3% vs 52.4%) and the peak of blood flow in stage with systole of right internal carotid artery (57.65±7.34 mm/s vs 52.41±7.97 mm/s) increased in patients of essential hypertension with cerebral ischemic group than those in patients of essential hypertension without cerebral ischemic group (P<0.05 or 0.01). The levels of bFGF in soft plaques group increased than that in flat plaques group (P<0.05). Conclusion Levels of bFGF and forming atherosclerosis may be an important factor during the development and discovery of cerebral ischemia, providing important basis for distingusihing the degree of desease with essential hypertension and preventing cerbral infarction, which contributes to intervene treatment.

Abstract:A20 was originally characterized as a TNF-inducible gene in human umbilical vein endothelial cells. As an inhibitor of NF-kappaB signaling, A20 could protect apoptosis, inflammatory and cardiac hypertrophy. We investigated the role of A20 on acute myocardial infarction (MI), ox-LDL-induced apoptosis in macrophage, and vascular remodeling. (1)We investigated the role of constitutive human A....

Abstract:C-reactive protein (CRP) is not only a predictor of cardiovascular events but also may be a new potential risk factor for the development of atherosclerosis. More recently, a number of studies using cultured cells showed that CRP can directly induce many cellular changes including the enhancement of cytokine and matrix metalloproteinases (MMP) production, cholesterol uptake by macrophages, ....

Abstract:近30年来糖尿病患病率在多数欧美和亚洲国家中迅速增高。糖尿病心血管并发症是糖尿病患者的主要死亡原因,其发生机制不甚明了。糖尿病有关的代谢因素增加循环和组织中氧化和糖化蛋白或脂蛋白的水平和氧化应激性。这些改变可引起动脉粥样硬化,凝血因子和血小板活性增高,纤溶活性减低,进一步导致血栓形成和急性组织缺血。纤溶酶原激活物抑制剂1(PAI-1)是一种急性反应物和炎症介质。血浆PAI-1增高被认为是一个内皮细胞功能失调的标....

Abstract:Aim To observe the inhibition effect of Tongxinluo on inflammation and intima proliferation response induced by interleukin-1β(IL-1β) in small swine,and to investigate its action mechanism. Methods Chinese mini-swine were randomly divided into three groups,control group,model group and Tongxinluo group.The control group and model group were fed with special diet;the Tongxinluo group were fed with special diet and Tongxinluo [1 g/(kg·d)].After 2 weeks' feeding,chest operation was done by isolation of left-anterior descending banch and circumflex branch proximate.Coronary artery membranna externa in swine was packed by paper towel absorbing saline agarose particle suspension in control group.But in model group and Tongxinluo group,coronary artery membranna externa was packed by paper towel absorbing IL-1β(2.5 μg)agarose particle suspension.After 2 weeks,lumens narrow was observed by coronary arteriogramphy.The vascular section in the treated site was taken out to be examined by pathology and the monocyte chemoattractant protein-1(MCP-1),P-selection,E-selection, intercell adhesion molecule-1(ICAM-1) and vascular cell adhesion molecule-1(VCAM-1) mRNA expression was measured by reverse transcription polymerase chain reaction analysis(RT-PCR). Results The coronary arteriogramphy showed that atherosclerotic changes and luminal stricture in local blood vessel in Tongxinluo group was dscreased compared with control group.The pathological examination showed that the degree of luminal stricture,the macrophage accumulation and endometrial hyperplasia in Tongxinluo group was obviously lighter than that of model group.The expression of mRNA of MCP-1,P-selection,E-selection,ICAM-1 and VCAM-1 in Tongxinluo group were decreased. Conclusions Tongxinluo could inhibit coronary artery early inflammation response and intima proliferation induced by interleukin-1β by inhibiting the mRNA expression of MCP-1,P-selection,E-selection, ICAM-1 and VCAM-1.

Abstract:Aim To determine the consequences of hypoxia inducible factor-1α(HIF-1α)-modified endothelial progenitor cells(EPC) on neovascularization in vivo and discussed the possible mechanisms of homing. Methods Adenovirus mediated human HIF-1α(Ad-HIF-1α) were transduced in human EPC in vitro.To determine if such transduced EPC may facilitate therapeutic neovascularization,heterologous EPC transduced with adenovirus encoding HIF-1α were administered to Balb/c nude mice with hindlimb ischemia.Exogenous EPC homing was observed and the possible mechanism of EPC homing was explored. Results Adenovirus mediated green fluorescence protein(GFP) was consistently expressed in EPC.A significantly higher capillary/myocyte ratio in mice transplanted with Ad-HIF-1α-EPC than in those receiving Ad-GFP-EPC at each time point was found.HIF-1α mRNA and protein expression in the ischemia zone was enhanced(p<0.05).Stromal derived factor(SDF) and CXCR4 upregulation were accompanied by paralleled vascular endothelial growth factor(VEGF) upregulation(p<0.05). Conclusions In vivo,gene-modified EPC facilitate the strategy of cell transplantation to augment naturally impaired neovascularization in an animal model of experimentally induced limb ischemia.The recruitment of EPC may be due to the inducement of SDF,CXCR4 and the VEGF induction may contribute to the EPC proliferation.

Abstract:Aim To detect the change of transcriptional profiling of C₂C₁₂ cells overexpressed krüppel-like factor 4(KLF4). Methods cDNA chip containing 14000 mouse cDNAs were employed to investigate the expression of genes influenced by KLF4 overexpression in C₂C₁₂ cells;bioinformatics database and software such as MEDLINE and Genomatix were performed to analyze the above genes. Results Two hundred and fifity genes were down-regulated in C₂C₁₂ cells overexpressed KLF4,with 155 known; Whereas 355 genes were up-regulated,with 255 known.Several genes about inflammatory or apoptosis were included. Conclusion Many downstream genes were influenced by KLF4 overexpression in C₂C₁₂ cells.

Abstract:Aim To study the effects of diabetes and balloon injury on diet induced arterial atherosclerosis in rabbit. Methods Set up rabbit atherosclerotic model by high cholesterol diet and injury of abdomen aorta's endothelium with balloon;half of the animals were injected alloxan one week before endothelium injury to make diabetic atherosclerotic model,atherosclerotic rabbits were injected physiological saline instead.Ten weeks after the surgery rabbits were sacrificed by euthanasia,transverse sections of the aorta were taken for H&E stain and immunohistochemistry to analyze intima incrassaition,quantities of macrophages and smooth muscle cells. Results Thoracic and abdomen aorta of both diabetic and non-diabetic rabbits developed atherosclerosis,and the ratio of intima to media thickness in thoracic aorta(0.9±0.18 and 0.7±0.2 repectively) was much lower than in abdomen aorta(6.9±2.0 and 6.9±2.1 repectively).Though intima thickness of the abdomen aorta was similar between the two groups,diabetic atherosclerotic rabbits had more macrophages(percentage of positive stain area in the intima was 39.7±8.7) and fewer smooth muscle cells(percentage of positive stain area in the intima was 29.6±5.5) in its intima than non-diabetic atherosclerotic rabbits(percentage of positive stain area of macrophages were 38.7±5.8 and that of smooth muscle cells were 35.2±4.9),especially at luminal 1/2 of intima(percentage of positive stain area of macrophages were 18.8±4.8 and 33.9±6.2 respectively and that of smooth muscle cells were 32.8±5.8,25.4±3.8 respectively). Conclusion Balloon injury of the endothelium accelerates atherosclerosis induced by diet in diabetic and non-diabetic rabbits;diabetes promotes inflammation of the atherosclerosis.

Abstract:Aim To evaluate the effect of hypoxia-inducible factor-1α(HIF-1α) to myocardial ischemia-reperfusion injury of rats. Methods Rats received ligation of the proximal left anterior descending coronary artery(LCA) for 30 min,followed by reperfusion for 180 min.They were divided into three groups: dimethyloxalylglycine(DMOG) group,control group and the sham operation group without ligation of LCA.In control group,0.5 mL saline were administered intraperitoneally 24 h before ischemia-reperfusion injury. Results DMOG increase the expression of HIF-1α and HO-1 gene significantly.The infarct size in DMOG group and saline group was 23.56%±2.12% and 35.21%±2.34% respectively.The infarct size in DMOG group decreased significantly than that in saline group(p<0.01).Caspase3 protein expression in DMOG group significantly decreased than that in control group(p<0.01).Plasma IL-8 level was significantly higher in saline group than that in sham operation group. Conclusion HIF-1α activation can increase the expression of HIF-1α and HO-1 gene and attenuate ischemia-reperfusion injury.

Abstract:Aim To investigate the protective effects and mechanisms of alendronate on vascular calcification in calcitoroil-induced rats with subtotal nephrectomy. Methods 30 SD rats were divided randomly into sham operation group;calcification group;alendronate group.Subtotal nephrectomy was performed by excision of two thirds of the left kidney followed by right nephrectomy 1 week later.Sham operation was performed by segregation of renal fascia without nephrectomy.After operation,all animals were treated with the high calcium(4%) and high phosphate(1.8%) diet.Meanwhile,all animals were intragastric administrated with calcitriol [1 μg/(kg·d)].Additionally,alendronate group rats were intraperitoneal injected with alendronate [0.25 mg/(kg·d)].4 weeks later,all rats were sacrificed by bleeding.For histological analysis of calcium accumulation,the beginning 1 cm of the abdominal aorta sections were removed and fixed in formalin,sectioned and histological stained(H&E and Von Kossa),and studied by microscope.For measurement of calcium content of the vessel wall,the abdominal aorta section beginning 1 cm and ending at femoral bifurcation were placed to different 2ml epitubes and 1 ml 150 mmol/L HCl was added to each tube.Each tube was closed securely and mixed end over end for 24 h.Calcium levels in the acid extract of tissues were determined by automatic biochemistry detection equipment.Serum concentration of osteocalcin was determined using an enzyme linked immunosorbent assay(ELISE). Results Extensive calcification was witnessed on the aortic tunica media of the calcification group rats,but less calcification in the alendronate group,and no calcification in the sham operation group.Calcium content of the vessel wall was significantly lower in alendronate group than those in calcification group(18.82±11.07 μmol/g vs 113.67±33.32 μmol/g,p<0.001),and there was no significant difference between alendronate group and sham operation group(18.82±11.07 μmol/g vs 6.67±2.82 μmol/g,p>0.05).Serum concentration of osteocalcin was significantly higher in alendronate group than those in calcification group(1.71±1.53 mmol/L vs 0.80±0.15 mmol/L,p<0.05),and sham operation group(1.71±1.53 mmol/L vs 0.74±0.38 mmol/L,p<0.05). Conclusion Injecting alendronate can inhibit vascular calcification in calcitoroil-induced rats with subtotal nephrectomy.

Abstract:Aim To assess the effects of aspirin on early atherosclerosis. Methods 24-month-old miniature pigs were randomly assigned into three groups: control(n=6) which were fed on normal chow,3.0% cholesterol group(n=8) which were fed on atherogenic diet containing 3.0% cholesterol,aspirin group(n=8) which were added aspirin 150 mg/d in 3% cholesterol group atherogenic diet.The atherogenic diet was continued for 6 months.Half pigs for each group were killed to determine the extent of atherosclerosis and the effects of aspirin after 4 months and 6 months. Results Cholesterol feeding induced significant artery atherosclerosis by 4 months.Morphometric analysis showed the maximal intima thickness,intima area were 107.2±45.7 μm and 0.113±0.05 mm2 in 3% cholesterol group,74.2±24.7 μm and 0.093±0.05 mm2 in aspirin group in 4 months.Whereas at the end of 6 months,maximal intima thickness,ratio of intima/media thickness,intima area and ratio of intima/media area were 152.9±86.1 μm,0.85±0.49,0.188±0.207 mm2 and 0.235±0.249 in 3% cholesterol group,47.8±19.8 μm,0.33±0.09,0.017±0.012 mm2 and 0.030±0.019 in aspirin group.The intima thickness,intima area had no statistic decline in 4 months but markedly declined in 6 months in aspirin group compared with 3% cholesterol group(p<0.05 or p<0.01). Conclusion Aspirin remarkably attenuated atherosclerosis plaque.

Abstract:Aim To study dynamic changes of serum von Willebrand factor(vWF) and high sensitivity C-reactive protein(hs-CRP) in renovascular hypertensive rats,and to investigate the effect of vWF and hs-CRP in the deployment of atherosclerosis(As) and stroke. Methods vWF and hs-CRP were measured with enzyme linked immunoabsorbent assay(ELISA),the results were compared with the control group and sham-opetated group. Results At the end of 4,8,12 weeks after the operation,the concenrations of serum vWF and hs-CRP were significantly increased compared with the control group and sham-operated group;the blood pressure were increasing in the hypertensive group after the operation,and the concenrations of serum vWF and hs-CRP were increasing simultaneously. Conclusion When the blood pressure were increasing,the concenrations of serum vWF and hs-CRP were increasing simultaneously;vWF and hs-CRP played vital roles in As.The high concenrations of serum vWF and hs-CRP might be the risk factors to ischemic stroke.

Abstract:Aim To clarify the effects of angiotensin Ⅱ on the calcification in rat vascular smooth muscle cells(VSMCs) and its signal transduction. Methods Calcification of cultured rat VSMCs was prepared by incubation with β-glycerophosphate.Calcification was confirmed by Von Kossa staining,and calcium content, alkaline phosphatases activity,osteocalcin and core binding factor alpha 1(Cbfa1) mRNA expression were measured. Results Angiotensin Ⅱ enhanced the increase of calcium content,alkaline phosphatases activity,osteocalcin concentration and Cbfa1 mRNA expression in calcified VSMCs(p<0.05).Angiotensin Ⅱ type 1 receptor blocker valsartan and selective protein kinase C(PKC) inhibitor inhibited the effects of angiotensin Ⅱ on vascular calcification(p<0.05). Conclusions These data suggested that β-glycerophosphate-induced calcification in VSMCs was enhanced by angiotensin Ⅱ through angiotensin Ⅱ type 1 receptor and PKC-Cbfa1 pathway.