Abstract:Aim To explore whether Na + H + exchanger (NHE) modulated atherogenesis, we investigated the effects of cariporide, a selective inhibitor of Na +- H + exchanger, against atherogenesis in animal models induced by high fat plus high cholesterol (high lipid) diet. Methods 22 healthy New Zealand rabbits were randomly divided into 3 groups: one group was fed normal chow (n=6), another group was fed high lipid diet chow served as atherogenesis control (n=8), the third group fed high lipid diet chow plus orally cariporide (0.1 mg/kg everyday) served as drug treated group (n=8). The blood was drawn from the central ear artery to assay serum lipoprotein at the prior experiment and animal sacrifice. Animals were killed in tenth week; the samples of arteries, lives and hearts were taken for morphologic analysis. Results The ultrastructures of aortic arteries showas follows. In normal diet group, the vascular intima were smooth and intact. In high lipid diet model group, the intima were become rough and thicker. A lot of lipoid foam cells migrated to regions of intima and smooth muscle cells (SMC) which associated injuries of internal elastic lamina. In the cariporide treated group, above described vascular injuries were significantly ameliorated, but it also was found that a few of foam cells scattered under intima of artery in a few cases (one rabbit). TC and low density lipoprotein cholesterol (LDLC) was significantly increased in both groups. Cariporide did not black increases of serum TC and LDLC levels. Conclusion Cariporide significantly decreased atherogenesis induced by hyperlipidemia. The actions of cariporide against atherogenesis were independent decreasing lipemia and mainly located in blood vessels and tissue.

Abstract:Aim To dissect the contribution of each repeat of the ligand binding domain of the VLDL receptor to ligand binding. Methods Several VLDLR recombinants lacking different repeat(s) were constructed by the method of oligonucleotide directed mutagenesis and then introduced into CHO cells by transfection. The ability of the mutant receptor to bind with β VLDL was measured. Results The result of the binding experiments showed that repeat 1 and 2 are the most important repeats required for β VLDL binding. Conclusion These results lay the foundation of VLDLR structure and function study.

Abstract:Aim To investigate the effect of nitric oxide (NO) on the apoptosis of cultured rat vascular smooth muscle cells (VSMC) and the role of focal adhesion kinase (FAK) in VSMC apoptosis induced by NO. Methods Flow cytometry, DNA gel electrophoresis, Northern blot and Western blot were used to determine the effect of different level of NO on VSMC apoptosis and to explore the relationship between VSMC apoptosis and expression activity of Bcl 2 and FAK genes. Results The results showed that both endogenous NO synthesized by VSMC and exogenous NO released by sodium nitroprusside (SNP) could significantly induce apoptosis of cultured rat VSMC and the apoptosis rate of VSMC was correlated with NO content in the medium. It is demonstrated that the expression activity of Bcl 2 and FAK genes markedly decreased during apoptosis development of VSMC induced by NO. Conclusions It is suggested that FAK may be involved in signal conduction of VSMC apoptosis induced by NO, and VSMC apoptosis was related to down regulation of Bcl 2 and FAK expression.

Abstract:Aim To understand whether native very low density lipoprotein (n VLDL) and oxidized very low density lipoprotein (ox VLDL) induce the cultured human umbilical endothelial cells (EC) to express macrophage inflammatory protein 1α (MIP 1α). Methods The EC were cultured in serum free medium containing 80 mg/L of n VLDL and ox VLDL respectively, while the control group without lipoproteins. After 24 h incubation, the EC grown on the cover slips were fixed and in situ hybridized with the Digoxigenin labeled MIP 1α cDNA probe. The MIP 1α mRNA expressed by the cells of every group was determined by reverse transcription polymerase chain reaction (RT PCR) as well. The MIP 1α protein expressed by the cells was detected by immunocytochemistry. Results Densitometry scan showed that the cultured EC could express MIP 1α mRNA, and the integral absorbance (A) values of the cells exposed to lipoproteins in situ hybridized with the probe metioned above were significantly higher than that of the control group. Analysis of variance demonstrated that there was significant difference among groups (p<0.01). RT PCR also showed that the A values of the MIP 1α mRNA expressed by the cells in n VLDL and ox VLDL group were 15.4 and 14.2 fold as much as that of the control group. Immunocytochemistry showed that the A values of the MIP 1α protein expression in the cells (brown granular substance) in n VLDL and ox VLDL group were significant higher than that of the control group. Analysis of variance showed a significant difference among groups (p<0.05). Conclusion n VLDL and ox VLDL were able to induce MIP 1α mRNA and protein expression in human umbilical vein endothelial cells at a high level, and may play an important role in atherogenesis through enhancing recruitment of monocytes to the intuma.

Abstract:Aim To investigate the effect of all trans retin oi c acid (ATRA) on reendothelialization and neointima formation in the rat's abdom inal artery after bal loon withdrawal injury. Methods The rat denuded ab dominal artery model with ba lloon withdrawal injury was performed. Twelve wistar rats were randomly assigne d to control group (intraperitoneal injection of 1 mL/d intralipid) and ATRA g roup [intraperitoneal injection of 4 μg/(g·d) ATRA] dissolved in intralipi d. After fourteen days balloon withdrawal injury in the situ perfusion fixa tion was performed with 10% formalin and the specimens were obtained. Histologi c observations and morphometric analysis were made. Results Reendothelialization area of arterial balloon injury increased in ATRA g roup compared with that in control group, while Neointima area is smaller in ATR A group than in c ontrol group. Conclusion ATRA has accelerated reendothelia lization and inhibited neointima formation of injuried vessels.

Abstract:Aim To investigate the protection of effective e lements from Scute Llaria Barbata on arterial endothelium. Methods The Wistar rats were fed with high cholesterol diet and treated with the A001 eleme nt and the B001 element of S cute Llaria Barbata for 60 days. The levels of the blood total cholesterol (TG) , triglycerides (TG), endothelin 1(ET 1), endothelial nitric oxide synthase (e N OS) and the positive rate of ET in "en face" preparation of arterial endotheli um were detected. Results The concentrations of TC and TG did not reduce in hyperlipidemic rats treated with both B001 and A001 . The blood ET 1, positive rate of ET in arterial endothelium and the morpholog ical injury of the arterial endothelium decreased significantly (p<0.05) and the blood eNOS increased significa ntly (p<0.05) in the hyperlipidemic rats treated with B001. The hyperlipidemic r ats treated with A001 did not show the same changes. Conclusion The B001 is the effective element of Scute Llaria Barbata, which can protect the arterial endot helium and impede the development of atherosclerosis.

Abstract:Aim To investigate whether oxidized low density l ipoprotein induces apoptosis of vascular smooth muscle cells. Methods The cultured porcine aortal smooth muscle cells incubated with oxidized low density lipoprotein of 15 mg/L for 72 hours in vitro. Then the samples were analyzed by fluorescence microscope?confocal microscope system and flow cytometry. Results Apoptosis was triggered by incubated with oxidized low density lipoprotein accompanying with apparently morphological changes. Conclusion In the process of which oxid ized low density lipoprotein induced vascular smooth muscle cells apoptosis, the abnorm al metabolism of cellular cholesterol may be have relation to apoptosis.

Abstract:Aim Toinvestigatewhetherstatinsaltertheexpressionoflectin likeoxidizedlowdensitylipoproteinreceptor (LOX) 1mRNA anduptakeofoxidizedlowdensitylipoproteinonculturedhumanumbilicalveinendothelialcells (hUVEC) . Methods TotalRNAwasextractedwithTrizolreagent. Reversetranscriptionpolymerasechainreaction (RT PCR)wasusedto quantifymRNAexpressionofLOX 1inhUVEC . Ox LDLwasradioiodinatedwith12 5 Itodetecttheuptakeofox LDLbyendothe lialcells. AspectrophotometricenzymeassaywasperformedtodeterminetheactivityofLDH . Results LOX 1mRNAexpressionwassignificantlysuppressedto 84 % ,6 9%and 4 8%ofcontrolafterhUVECwereincubated with 1,10 ,5 0 μmol/Loflovastatinfor4 8h. LOX 1mRNAexpressionwassignificantlysuppressedto 82 % ,6 9%and 5 2 %of controlinthecellstreatedwithlovastatinafter2 4 ,4 8or 96hrespectively . Conclusion LovastatinreducestheexpressionofLOX 1mRNAanduptakeofox LDLwithoutanytoxiceffectswhichmaycon tributetotheantiatheroscleroticpotentialoflovastatin .

Abstract:Aim To clarify the binding and degradative pathwa y of lipoprotein(a) and oxidized lipoprotein(a). Methods The binding and the competitive binding assays about Lp(a) and oxidized Lp(a) with the mouse peritoneal macrophage were performed. Results Lp(a) was bound with affinity, saturation to macro phage surface. LDL was minimal potency in competing with Lp(a) for the binding site, in contrast, ox LDL and ox Lp(a) were efficient competitor for the bindi ng of Lp(a). Oxidative modification of Lp(a) by Cu 2+ increased markedly i ts binding to macrophages. Ox Lp(a) and ox LDL were excellent competitor for the binding of ox Lp(a) to macrophages. Conclusions The binding of Lp(a) with macrophages by the scavenger receptor medi ated pathway; Scavenger receptor provided a significant route for the metabolism of ox Lp(a), perhaps, any other special receptor also mediated the binding of ox Lp(a) to macrophages.

Abstract:Aim To observe the influence of flu vast atin and simvastatin on atherogenesis and expression of vascular cell adhesion m olecule 1(VCAM 1) on aortic intima in ApoE deficient mice in order to investi gate the non lipid mechanisms of 3 hydroxy 3 methyl glutaryl coenzyme A(HMG CoA) reductase inhibitors on anti atherosclerosis. Methods Under the general feeding, pravastatin (every day 10 mg/kg) and sim vastatin (every day 5 mg/kg) were injected into the stomach of ApoE deficien t mice for 4 weeks . The areas of atherosclerotic plaques, and ratio of plaque area to aortic lumi nal area were detected by histochemistry technique. The expression of VCAM 1 o n aortic intima in ApoE deficient mice was detected by immunohistochemistry and Western blotting. Results Compared with controls, both of pravastatin and simvastatin could delay plaque formation in ApoE defic ient mice and diminish plaque size, and could inhibit VCAM 1 expression on aort ic intima, which were inconsistent with their cholesterol lowering effect. The inhibition of VCAM 1 expression by pravastatin or simvastatin in mice of 10 or 20 weeks old was slightly stronger than that in 30 weeks old mice. There was n o significant difference in inhibition of plaque information between pravastatin and simvastatin in view of the dosages used in this study. Concl usions The inhibitory effect of pravastatin and simvastatin on at h erogenesis of sclerotic lesions may crucially contribute to the clinical benefit s of HMG CoA reductase inhibitors on coronary artery disease. The inhibition o f VCAM 1 expression by pravastatin or simvastatin on aortic intima in ApoE def icient mice may play an important role in the reduction of VCAM 1 dependent mo nocytes adhesion to endothelium and reflect a new mechanism of statins on anti atherosclerosis, which is probably independent of lipid regulation.

Abstract:Aim The present study was to investigate the effe cts of Fluva statin on vascular smooth muscle cells (VSMC) proliferation in atherosclerotic model of iliac artery of rabbit after endothelial denudation. Method s The New Zealand White Rabbits were randomized into Fluvastatin group (n=28) and control group(n=28). The atheroscl erotic lesions were developed by inducing endothelial denudation with balloon ca theter and being given high dosag e of cholesterol diet at the same time. The experimental animals were killed an d iliac arteries were removed at 1, 2, 4, 8 week after endothelial injury respect i vely. Through the methods of routine immunohistochemical staining and image ana l ysis system, we observed the effects of Fluvastatin on the expression between pr oliferating cell nuclear antigen (PCNA) and platelet derived growth factor BB (PDGF BB) in neointima. Results Percentages of PCNA postiv e cells in the intima of Fluvastatin group was lower than that in control group apparently. The content of PDGF BB in Fluvastatin group was al ways at a lower level and even lower than that in control group (p<0.05). Coefficient of linear correlation of PCNA with PDGF BB was 0.7127( p<0.01). Conlusion Fluvastatin may partly in hibit VSMC proliferation through decreasing the synthesis of PDGF BB.

Abstract:Aim To elucidate whether ischemia and reperfusi on can induce cardiomyocytes apoptosis and the relation between polymorphnuclear n eutrophils (PMN) infiltration and cardiomyocytes apoptosis. The effect of ische mic preconditioning on apoptosis was also investigated. Methods Forty eight male Wistar rats were ra ndomly assigned to one of the following four groups: I+1h R group, I+2h R group , I+3h R group and IP group. The following variables were measured: cardiac fu nction, infarct size, PMN counts, myocardial myeloperoxidase activity and ap optotic cell in dex. Results Ischemia and reperfusion caused obvio us cardiac function injury, PMN infiltration and cardiomyocytes apoptosis, and heart injury increased as the reperfusion continued; ischemic preconditionin g significantly reduced heart injury and apoptosis. Correlative analysis betwee n PMN count and apoptotic index suggested a significant positive correlation ex isted between PMN infiltration and cardiomyocyte apoptosis (r=0 .943,p<0.001). Conclusion Ischemic pr econditioning significantly reduced PMN infiltration and attenuated myocyte apop tosis.

Abstract:Aim To investigate the relationship between age a nd the development o f atherosclerosis. Methods The mictroarchitecture of intima at apical and later al zones was studied respectively at branch regions of human abdominal aorta in f erior mesenteric artery using transmission electron microscopy (n=30, age ranges from 1 day to 77 years). Results With aging, the th ickness of intima had no a pparent changes at apical zones, where smooth muscle cells turned into constract ile phenotypies and decreased in number, without atherosclerotic lesions; While the thickness of intima increased significantly, where smooth muscle cells remai ned in synthetic states and slightly increased in number, with some atherosclero tic lesions such as lipid deposition, edema, et al. Conclusions The intima at lateral zones has some distinct structural peculiarities with age, which is morp hological basis of atherosclerosis. That is likely caused by hemodynamic status there.

Abstract:Aim To identify, clone and sequence Chinese ma ture Phospholipicl Transfer Protein (PLTP) peptide gene. Methods Total RNA was extracted from Chinese fetal liver tissue, cDNA fragment encoding human PLTP was amplified by RT PCR using specific primers, and then was cloned into pGEM T vector. Inserted PLTP gene was sequenced by ABI 377 DNA Sequenc er. Results Chinese PLTP gene was successfully cloned. Conclusion Analytical result indicates it is 1 428 b ase pairs in length and has 100% nucleotide homology with that reported previous ly.

Abstract:Aim To investigate the changes of plasma homocyst eine and lipid before and after therapy. Methods Simavatatin tablet s and/or Lipanthyl tablets were used to treat hyperlipidemic patients. Folate and Vitamin B 12 were used to tr eat hyperhomocysteinemia. Plasma homocysteine and lipid were detected before an d after treatment. Results After treatment of hyperl ipidemic patients, the decrease of lipid was significant but the decrease of hom ocysteine was not. After treatment of hyperhomocysteinemia, the decrea se of homocysteine was significant whilc the decrease of lipid was not. Conclusions With the treatment of hyperlipimia using Simava tatin tablets and Lipanthyl tablets, the plasma lipid decreased significant ly, but the plasma homocysteine decreased nonsignificantly. The plasma hom ocysteine decreased strongly by using Folate and Vitamin B 12 , but the chan ges of lipid were not significant.

Abstract:Aim To determine whether Xuezhikang, a HMG CoA reductase inhibito r made in China, affects CD11b expression and adhesiveness of monocytes to endot helial cells in vitro after the treatment of patients with hypercholesterolemia. Methods Fifteen patients with hypercholesterolemia were treated with Xuezhikang (1.2 g/d) for 4 weeks. Isolated human blood monoc ytes were subject to flow cytometric detection of CD11b and adhesion assays to l ive human endothelial cells were made before and after the treatments of Xuezhik ang. Results The average fluorescence intensity and the relative fluorescence intensity of CD11b expressed in monocytes and lymphocy tes from the peripheral venous blood were lowered after the treatment of Xuezhik ang. The expression rate of CD11b in lymphocytes was decreased from 27%±4% to 13%±3% (p<0.05,n=15). The adhesion rate of mo nocytes to endothelial cells was reduced from 5.5%±2.3% to 1.8%±0.8%(p<0.05,n=15). Pretreatment of CD11b mAb in vitro ca n inhibit the adhesion of monoctyes to endothelial cells. Conclus ions Xuezhikang, a kind of HMG CoA reductase inhibitor, can inhi bit the expression of CD11b in monoctyes and the adhesion of monocytes to endoth elial cells with the cholesterol loweing effects. The nonlipid mechanisms may involve in it.

Abstract:Aim To determine the relationship between hyper uricemia and coronar y artery disease (CAD). Methods Serum concentrat ion of uric acid was measured and other biochemical assessments were made at a fasting state in 2 009 patients who have undergone coronary angiography. Results S erum uric acid level was significantly higher in CAD group than in normal corona ry group(369±97 mmol/L vs. 356±94 mmol/L, p<0.01). Uni var iable analysis with Mantel Haenszel Chi test in dicated that hyperuricemia had statistically significant relation with CAD ( χ 2 MH =4.364, Or=1.24, P=0.037). Despite similar mean value of serum uric acid level between acute myocardial infarction (AMI) and nor mal coronary groups, the number of patients with abnormal serum uric acid level was greater in the former than in the latter (p<0.05). Logistic regression analysis with control of other covariables showed a significant rela tionship between plasma uric acid level and CAD (χ 2 wald =4.76, P=0.0292, Or=0.999) and AMI (χ 2 wald =23.48,P= 0.0001,Or= 1.004). Conclusions Serum uric acid level had a strong relationship with the i ncidence of CAD. This relation was further enhanced when hyperuricemia is associ ated with diabetes mellitus and cigarette smoking.

Abstract:Aim To investigate the relationship between cor onary artery disease and hypertriglyceridemia? hypo high density lipoprotei nemia. Methods 54 cases with stable angina pectoris or old myocardial infarction were divided into four groups: 15 cases in non major risk factor group, 11 cases in hypertriglyce ridemia and hypo high density lipoproteinemia group, 15 cases in diabetes mel litus g roup, and 13 cases in diabetes mellitus and hypertriglyceridemia and hypo high densit y lipoproteinemia group. Some risk factor such as smoking years, blood glucos e, systolic blood pressure, choleserol, low density lipoprotein cholesterol(LDLc ), trig lyceride, high density lipoprotein cholesterol (HDLc) were measured. The rel ationship between above factors and the degrees of coronary artery lesion with a ngiography were detected. Results The coronary arter y stenosis s core was significantly higher in diabetes mellitus and hypertriglyceridemia, and hypo high density lipoproteinemia group (10.5±3.4) than in other groups (p<0.05), the B 2?C type of coronar y artery disease in diabetes mellitus and hypertriglyceridemia and hypo high d en sity lipoproteinemia group were significantly more than in non major risk fac tor group (p<0.05), and three vessel diseases increase more in this group; the lineal regression relationship were found between blo od glucose, triglyceride, HDLc and coronary artery stenosis score with stepwise regression analysi s. Conclusions The higher the levels of blood gluc ose and triglyceride become, the lower the level of HDLc. The coronary artery s tenosis score will become higher, with the degree of coronary artery lesion be coming heavier.

Abstract:Aim To study the effect of fluvastatin on serum l ip id and adhesive molecules. Methods 58 patients wit h hyperlipidemia (male 35, female 23) received fluvastatin 40 mg, po, for one week and then the dosage were reduced to 20 mg night. The therpeutic course wa s 6 8 week. Results After treatment the average lev els of serum TC, TG, LDL, soluble intercellular adhesion molecule (sICAM) 1 and soluble vascular cell adhesion molecule (sVCAM) 1 were lowered significantly (p<0.05), but HDL was not significantly elevated. Conclusion Fluvastatin not only decreased TC,TG and LDL, but als o decreased sICAM 1 and sVCAM 1 concentration.

Abstract:Aim To get a rapid, simple label method for lipop rotein , an improved method was established with the fluorescent probe DiI. Method The serum at the bottom of the post ultracentrifuged tu be, instead of lipoprotein deficient serum (LPDS), was used as the medium to app lying in lipoprotein labeling. Results Binding assay showed that the labeled lipoprotein can bind with macrophage and chinese hamster ovocyte (CHO) cells with typical saturation able model. The results p roved that present method labeling lipoprotein have the normal lipoprotein ligan d binding properties. Conclusion This method is sim ple, rapid and reliable. It could be applied widely in lipoprotein receptor s tudy.