Abstract:Aim To explore the molecular mechanisms of cultured rat vascular smooth muscle cell (VSMC) migration stimulated by newborn calf serum (NCS) or basic fibroblast growth factor (bFGF) and the mechanism of heparin inhibition on VSMC migration induced by NCS. Methods Rat osteopontin cDNA probe was amplified by RT PCR. After hydroxyurea inhibited VSMC proliferation induced by bFGF and NCS, osteopontin gene expression and cell energy exchange were respectively analyzed by Northern blotting and creatine kinase NAC kit. Results Northern blotting results showed that bFGF and NCS could induce VSMC osteopontin gene expression. However, heparin could inhibit osteopontin gene expression induced by NCS. Creatine kinase activity in VSMC stimulated by bFGFand NCS increased by 76% and 61%, respectively, as compared with that of the control (P<0.05), and creatine kinase activity of the NCS+heparin group was 22% lower than that of the NCS group. Conclusion VSMC migration stimulated by NCS or bFGF and heparin inhibition on VSMC migration induced by NCS were related with osteopontin gene expression and increased energy exchange.

Abstract:Aim To investigate the molecular mechanism of the Zhitai Capsule, which is a phelgm and stasis treating prescription, protecting vascular endothelial cells and against atherosclerosis. Methods 24 male New Zeland White rabbits were divided into four groups: the normal control group, the hypercholesterol group, the Gimfibrozil group and the Zhitai group. Four months later, the rabbits were killed and the aortas were fixed. Using NADPH diaphorase histochemistry and in situ hybridization, we have studied the activity and mRNA expression of nitric oxide synthase(NOS),as well as the mRNA expression of endothelin in aorta. Results The NOS activity in the aortas of Zhitai group (1.38±0.63) were significantly higher than that of hypercholesterol group(0.40±0.24)(P<0.01); that the NOS mRNA expression in the aortas of Zhitai group(1.38±0.48) were much higher than that of hypercholesterol group(1.00±0.00)(P<0.05); and that the NOS mRNA expression in the aortas of Zhitai group (2.43±0.53) were much lower than that of hypercholesterol group(3.33±0.82)(P<0.05). The Zhitai capsule can prevent the activity and mRNA expression of NOS in advanced atherosclerosis from decending, the mRNA expression of endothelin in advanced atherosclerosis from increasing, and decrease the lipid necrosis core. Conclusion Zhitai capsual’s antiatherosclerosis role was correlated with it′s effect on mRNA expression of nitric oxide synthase and endothelin.

Abstract:Aim To study the distribution of low density lipoprotein (LDL) subfractions and its susceptibility to in vitro oxidation in patients with chronic kidney failure (CKF). Methods LDL subfractions were isolated from plasma by non equilibrium density gradient centrifugation directly, and distribution of LDL subfractions in CKF patients were analyzed. The continuous monitoring of LDL oxidation by absorbance measurements at 234 nm was used to analyze the susceptibility of LDL and its subfractions to Cu 2+ mediated oxidation in patients with CKF. Results The frequency distributions of LDL subfractions in CKF patients had no different from that of control. The lag time in the test for LDL oxidizability in CKF patients was shorter than that in control (48.2±24.7 vs 93.3±37.2 min, P<0.05). The degree of oxidized LDL was correlated with the lag time of LDL oxidizability. The total amount of conjugated dienes and the maximal rate of oxidation had no difference. Conclusions Circulating LDL isolated from CKF patients is more susceptible to oxidation in vitro than that of controls, this may be associated with the level of oxidative modification LDL.

Abstract:Aim The antioxidation effect of resveratrol, a polyphenolic compound in red wine, on the oxidation of human low density lipoprotein (LDL) were systematically investigated by two different oxidation systems. Methods Oxidation of LDL was induced by adding Cu 2+ or azo compound. The extent of LDL modification was assessed by measuring the formation of thiobarbituric acid reactive substances (TBARS) and the relative electropherosis mobilities (REM) to native LDL. Results Treatment of LDL with resveratrol (50 μmol/L) reduced the production of TBARS and REM of LDL during Cu 2+ induced oxidation by 70.5% and 42.3%, respectively (P<0.01). The lag phase of LDL oxidation induced by copper ion or azo compound was delayed. Conclusion Resveratrol could protect LDL against both Cu 2+ induced and azo compound initiated oxidative modification in vitro, which might be contributed by its free radicals scavenging capacity.

Abstract:Aim To investigate the possible effect of sodium hydrogen antiporter on apoptosis of vascular smooth muscle cells induced by nitric oxide. Methods Rats’ vascular smooth muscle cells were used for experiment at passage 5 to 8. After reaching confluence, cells were subcultured on glass cover slides for observing under microscope, and cells were also subcultured on special Petri culture plate to examine intracellular pH through ACAS570. S Nitroso N acetylpenicillamine was used as nitric oxide donor in cell treatment. Results Intracellular pH in vascular smooth muscle cells was decreased by S Nitroso N acetylpenicillamine, but this effect was inhibited by amiloride, an blocker of of sodium hydrogen antiporter and amiloride could also inhibit the apoptosis in vascular smooth muscle cells induced by S Nitroso N acetylpenicillamine. Conclusion sodium hydrogen antiporter may take part in the apoptosis in vascular smooth muscle cells induced by nitric oxide donor S Nitroso N acetylpenicillamine

Abstract:Aim To study the role of ischemic preconditioning in reducing myocardial injuries and reducing myocyte apoptosis. Methods The models of ischemic preconditioning (IP), ischemia reperfusion injury (RI) and continuous ischemia (CI) were made with rabbits.The presence of apoptotic myocytes was demonstrated by the method of terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL). Also, the activities of plasma superoxide dismutase(SOD) were determined by the method of xanthine oxidase and the contents of serum malonic aldehyde (MDA) were checked by colorimetry. Their serum cardiac enzymes including aspartate transaminase (AST), lactate dehydrogenase(LDH), creatine kinase(CK)and hydroxybutyrate dehydrogenase(HBD) were checked at same time. Results The serum contents of AST, LDH,CK and HBD in RI and CI groups were higher than those in IP and control groups (P< 0.01), which meaned that there was severe damage in myocytes of RI and CI groups. Plasma activities of SOD in RI and CI groups decreased(P<0.01, compared with IP and control groups), but there was no statistic significance between IP and control groups. The contents of serum MDA in RI group was also higher than other groups (P<0.01 or 0.05). The rate of apoptotic myocytes in IP group (24.44±2.96%)was higher than that in control(0.71±0.51%), but was lower than that in CI (29.56±3.08%) and RI groups (43.33±4.92%) significantly (P<0.001). Conclusion This study suggested that IP could prevent myocardial injury induced by ischemia-reperfusion and continuous ischemia , and also reduce the rates of myocyte apoptosis in those situations.

Abstract:Aim To investigate whether the release of endogenous calcitonin gene related peptide (CGRP) during preconditioning ischemic insult played an important role in myocardial ischemic preconditioning (IPC) in the intact rat model. Methods Plasma CGRP concentration at the end of first or third ischemic insult was determined with radioimmunoassay. Infarct size as a percentage of the area at risk was determined with nitro blue tetrazolium. Results Mean plasma CGRP levels at the end of first and third ischemic insult were markedly increased in the IPC compared with control(P< 0.01), and it was markedly higher at the end of third ischemia than first in the IPC group (P<0.05). IPC markedly reduced the incidences of ventricular arrhythmias during 30 min ischemia and 2 h reperfusion, which were markedly attenuated by pretreatment with CGRP PcAb. There was a marked reduction infarct size in IPC group (P<0.01) which was markedly attenuated by pretreatment with CGRP PcAb (P<0.01). Conclusion Calcitonin gene related peptide plays an important role in myocardial ischemic preconditioning.

Abstract:Aim To investigate the effects of macrophage activation on the capacity of apolipoprotein AI to promote cellular cholesterol efflux. Methods 3H labled acetyl low density lipoprotein (ac LDL) was employed as an inducer for foam cell formation by thioglycolate were cultured with apo AI for 18 h, 3 H cholesterol released from cells to the medium was measured by scintillation spectrometry. Results The efflux of cholesterol from inflammatory macrophages to apo AI did not increase markedly with the increase of the concentration of apo AI as that observed in resident macrophages. The macrophage activation resulted in a significantly decrease in the capacity of apo AI (20 mg/L) to promote cellular cholesterol efflux (P<0.01), but had no effect on HDL. Conclusion The macrophages activation may effectively inhibit the capacity of apo AI to remove cholesterol from cells.

Abstract:Aim The effect of bovine aortic heparan sulfate proteoglycan (HSPG), chondroitin sulfate proteoglycan (CSPG), dermatan chondroitin sulfate proteoglycan (DSCSPG) and mixed PG on the growth of cultured human aortic smooth muscle cells (hASMC) was studied in this article. Methods PGs were isolated by dissociative extraction, ion exchange chromatography and gel filtration from bovine aortic intima media and their effects on the growth of cultured hASMC were studied by cell count and 3 H TdR incorpor...

Abstract:Aim To evaluate the apolipoprotein E (Apo E) genotype distribution in two type of stroke. Methods Polymerase chain reaction restriction segment length polymorphism (PCR RFLP) wae used to examined the Apo E genotype in 180 patients with hemorrhagic stroke (90) and ischemic stroke (90), along with the 107 age and sex matched controls. Results The frequency of Apo E ε2/ε3 was higher in hemorrhagic stroke group (28.9%) than in the controls (16.7%, P<0.05), while the Apo E ε3/ε3 frequency was lower in the cases (54.4%) than in the controls (70.0%, P<0.05). The Apo E ε3/ε4 was more frequent in the ischemic stroke group (21. 1%) than in the controls (8.9%, P<0.05). Conclusion Apo E ε2 may be related to the risk of hemorrhagic stroke and that Apo E ε4 may be an important risk factor for ischemic stroke.

Abstract:Aim To study on the three polymorphic sites of apolipoprotein B gene Ecoli, Mspl and Xbal and their association with atherosclerosis in Chinese Han nationality. Methods Subjects were 84 patients of myocardial infraction (MI) and 84 normal matched controls. Polymorphisms of apo B gene were determined by using polymerase chain reaction technique. The plasma lipid were performed, TC were measured by enzymatic method, TG were measured by ethylene acetone micro method, HDLC was determined after phosphotungstate precipitation, LDLC was measured by polyvinyl sulfate precipitation method, apoAI and apo B were quantified by immunonephrometry and the determination of Lp(a) was performed with ELISA (enzyme linked immunosorbent assay). Results In Chinese Han population, the appearance of E+ and M- were frequent ones at EcoRI and MapI restriction sites in apoB gene (the frequencies were 0.945 and 0.969 respectively); comparatively, relative frequencies of E- and M+ alleles were only 0.056 and 0.081 respectively, which were significantly lower than in the White. The relative frequency of X+ allele at Xbal restriction site in apoB gene was significantly higher in the MI group than that in the control group ( the frequencies were 0.0774 and 0.0297,(P<0.05). The levels of TC, TG, LDLC, apoB, Lp(a) and the ratio of LDLC/HDLC, (TC-HDLC/HDLC) were significantly higher; and the level of HDLC, the ratio of apoAI/B were significantly lower in MI patients as compared with those of controls (P<0.05).The incidence of hyperlipidemia in MI group was 72% higher than control group (P<0.05), But the three polymorphic sites of apo B were not associated with the variation of the levels of lipid, lipoprotein and apolipoprotien. Conclusion The data suggested that allele of X+ at Xbal restriction sites in apoB gene was associated with susceptibility to coronary atherosclerosis.

Abstract:Aim To study the effect of neferine (Nef) on copper mediated low density lipoprotein (LDL) and very low density lipoprotein (VLDL) oxidative modification. Methods Lipoproteins (LDL or VLDL) was isolated by ultracenfugation from normal human plasma. The extant of lipid peroxidation was measured in terms of thiobarbituric acid reactive substances (TBARS) express as malondiadelhyde (MDA) equivalents. The electrophoretic mobility of lipoproteins was determined on an agrose gel eletrophoresis. Mouse peritoneal macrophages (MP) was obtained by the method of Goldstein. Results Neferine inhibited the generation of TBARS in LDL and VLDL. Nef could decrease the electrophoretic mobility of VLDL, but it couldn’t do this to LDL. The oxidized low density lipoproteins (ox LDL) and oxidized very low density lipoproteins (ox VLDL) was more efficient than Cu 2+ + Nef + LDL and Cu 2+ + Nef+ VLDL in stimulating lipid accumulation in MP. Conclusions Neferine can inhibit copper mediated LDL or VLDL oxidation and by which inhibit lipid accumulation in MP.

Abstract:Aim To investigate enalapril, nifedipine and lovastatin had the effects on experimental atherosclerosis in cholesterol rabbits. Methods Thirty seven rabbits were divide into five groups accidentally. Group A received normal chow, group B received cholesterol, group C received cholesterol plus enalapril, group D received cholesterol plus nifedipine and group E received cholesterol plus lovastatin. All rabbits were fed for sixteen weeks. At the end of the study, all animals were killed to determine the aortas cholesterol concentration and the percentage of aortic lesions covering the intimal surface. Results Enalapril and nifedipine didn’t affect lipid level. Lovastatin reduced total cholesterol, triglyceride, and LDL level. The LDL/HDL ratio was less in lovastatin group than that in cholesterol, enalapril, and nifedipinegroups. The aortas cholesterol concentration and the plaque square in all drug groups were lower than in cholesterolgroup (P<0.05). Conclusion These three drugs had the effects of prophylaxis in experimental atherosclerosis.

Abstract:Aim To explore the relation between insulin resistance (IR) and deletion polymorphism of angiotensin converting enzyme(ACE)gene in patients with old myocardial infarction. Methods The insulin resistance was calculated by fast plasma insulin × fast plasma glucose÷22.5; The deletion (D)/insertion(I) polymorphism of the ACE gene was determined by polymerase chain reaction in 55 patients with old myocardial infarction and in47 control subjects. Results The insulin resistance level did not increase significantally in patients with old myocardial infarction, compared with control subjects(P>0.05); There were not significant difference of IR level among ACE DD、DI、Ⅱ gene types in the old myocardial infarction group(P>0.05 ). Conclusions The association of deletion polymorphism of ACE gene and IR with coronary heart disease were not found. The relation between deletion polymorphism of ACE gene and IR was also not identified in patients with coronary heart disease.

Abstract:Aim To investigate the relationship between apolipoprotein E (apo E) genotype and acute myocardial infarction(AMI) and the influence of apolipoprotein E phenotype on serum cholesterol metabolism. Methods Polymorphism of apolipoprotein E gene were detected by polymerase chain reaction fragmant length polymorphism (PCR RFLP) in a cross sectional study of 50 patients with AMI and 100 healthy subjects. The lipids of these subjects were measured with enzyme’s methods. Results Comparied with control group, the distribution of apolipoprotein E ε4 allele’s and genotype’s frequencies in AMI group were increased(P<0.05), patients with ε4 carrier had higher total cholesterol. Low density lipoprotein(LDL) cholesterol and apolipoprotein B concentrations than subjects with homozygous for ε3(P<0.05). Conclusion Apolipoprotein E polymorphism is associated with AMI and affects the metabolism of serum cholesterol.

Abstract:Aim To study the mechanism and effect of antioxidant vitamin E on advanced glycosylation end products (AGEs) stimulated signal transduction in cultured aortic smooth muscle cells (ASMC). Methods Separation and quantitative measurement of diglyceride (DG) were studied with thin layer chromatography, autoradiography and radioenzymatic assay. Results The arbitrary fluorescence unit (AFU) of AGEs was significantly decreased when Vit E were incubated with glucose and BSA. The level of DG, which was stimulated by the AGEs, was decreased. While Vit E were preincubated with ASMC for 4 h. The level of DG induced by AGEs was also lower markedly. Conclusion Vit E can signicantly inhibit the formation of AGEs. It can also markedly attenuate the accumulation of DG in ASMC.

Abstract:Aim To study the effect and mechanism of cardiovascular protection of 17 beta estrodial on coagulative and fibrinolytic system in animal model and cultured human umbilical vein endothelial cell. Methods The activity changes of tissue plasminogen activator (t PA) and plasminogen activator inhibitor (PAI 1) both in vivo and in vitro were measured by spectrophotometric assay, and the level of message RNA of them were examined by in situ hybridization. In addition, the level of fibrinogen in plasma was investigated by the instrument of automobile analysing coagulative and fibrinolytic system. The intracellular free calcium concentration in single human umbilical vein endothelial cell (hUVEC) was also determined by kinetic observation with Laster Confocal Microscope. Results In vivo, the t PA activity was decreased significantly in the group that the rats were ovariectomized for 5 weeks (OVX2 group) (P<0.05) and PAI 1 activity was increased significantly in both OVX1 (the rats ovariectomized for 3 weeks) and OVX2. Fibrinogen of plasma in OVX1 was also increased (P<0.05 vs Control). In the two groups of OVX treated with 17 beta estrodial, PAI 1 activity was decreased (P<0.01, respectively) and t PA activity was increased in OVX+17 beta estrodial (P<0.05). In situ hybridization showed PAI 1 mRNA in OVX1 tissue was higher than the control. In vitro, the t PA activity and gene expression were increased with the raising concentration of 17 beta estrodial. PAI 1 mRNA was changeless between different concentrations of 17beta estrodial. The concentration of intracellular calcium free of HUVEC was decreased in physical 17 beta estrodial. Conclusions The effect of cardiovascular protect of estrogen was associated with decrease in PAI 1, fibrinogen, and [Ca 2+ ]i , as well as increase in t PA activity and gene expression.

Abstract:Aim To investigate the influenece of β receptor blocker on Q T dispersion of old myocardial infarction. Methods Q T dispersion(Q Td),diameter of the left ventricular end diastolic phase(DLVED),ejection fraction (EF),paired ventricular premature complex(paired VPC),non sustained ventricular tachycardia(NSVT)were measured in 42 patients with old myocardial infarction(OMI)and 36 patients with hypertensive heart disease (HHD) as control,to investigate the variance of Q Td,paired VPC,NSVT after and before treatment with β blocker. Results Q Td,DLVED,EF were not significant difference (P>0.05) and paired VPC,NSVT were significant difference (P<0.05)between two groups of patients before treatment,significantly greater in OMI group than in control group.Q Td,paired VPC,NSVT were tended to decrease significantly (P<0.05)in OMI group and no significantly(P>0.05) in control group, after treatment with β recepter blocker than before. Conclusion These results suggest that treatment with β blocker may reduce Q Td in the recovery phase and malignant arrhythmia by reducing electrophysiological instability in old myocardial infarction.

Abstract:Aim The present study was to compare the lipid lowering efficacy of Luozhida (lovastatin) with zocor (simvastatin). Methods 63 subjects were randomized into two groups: 31 subjects in luozhida group, 32 subjects in zocor group as controls.Blood lipid levels were determined prior to lipid lowering treatment and at the end of 4 and 8 week treatment respectively. Results At the end of 8 week treatment, levels of TC, LDLC and apolipoprotein B in luozhida group and zocor group significantly reduced (P<0.01). The LDL cholesterol lowering efficacy of luozhida was significantly weaker than that of zocor. TG level was also significantly lower after treatment compared with that prior to treatment in both groups (P<0.05). At the end of 8 week treatment HDLC level tended to be higher in both group, though the difference was not statistically significant, while apolipoprotein AI level obviously increased in the luozhida group. There was no obvious side effects during the period of luozhida and zocor treatment and the patient’s tolerance was good. Conclusion Luozhida, as a domestically made lipid lowering drug,is effective for the treatment of hypercholesterolemia.

Abstract:Aim To study the relationship between coronary atherosclerosis and plasma endothelin. Methods The levels of plasma endothelin of 43 patients with coronary atherosclerotic heart disease(CHD) and 21 normal persons were measured using radioimmunoassay (RIA). The group with multi vessel disease and the group with single vessel disease were divided accoding to the numbers of vessel disease. Results The mean level of plasma endthelin was obviously increased in CHD group than in control group(P<0.01); The mean level of plasma endothelin was markedly increased in the group with multi vessel disease than in the group with single vessel disease (P<0.05); The concentration of plasma endothelin was strongly related to the number of vessel disease (r=0.414; P<0.05). Conclusion The plasma endothelin had correlation with the occurrence or degree of coronary atherosclerosis.

Abstract:Aim A method to determining CD36 antigen gene expression in periphery blood mononuclear leukocytes (PBMNL)is established. Methods The expression of CD36 antigen gene was measured by reverse transcription polymerase chain reaction (RT PCR)with β actin as the internal standard. RT PCR products were quantified by gel scanning. Results The gel electrophoresis revealed that the molecular weight of CD36 antigen RT PCR products was 390bp, that of β actin was 540bp. It is a stable method that CD36 antigen gene expression was expressed by CD36 antigen/β actin . The gene expression was significantly enhanced in the patients with carotid atherosclerosis. Conclusion RT PCR is an ideal method to determine expression of CD36 antigen gene, which might be used in the study of atherosclerosis.

Abstract:Aim To establish a method for low density lipoprotein receptor (LDLR) genotyping and research the relationship between genetic polymorphism of LDLR and variation in serum lipid levels. Methods The frequency distribution of LDLR, apo E genotypes and aelles were analysed in the method for LDLR, apo E genotyping using polymerase chain reaction restriction fragment length polymorphism (PCR RFLP), in samples of 81 unrelated normallipidaemic individuals and 51 hypercholesterolemia patients. And to examine the interaction effect on lipid level between the LDLR/AvaⅡ and apolipoprotein E polymorphisms. Results There is a significant relationship between“ + ”allele and high total cholesterol, LDL cholesterol levels. The apo E 3/2 and LDLR AvaⅡ(-/-) individual’s cholesterol level was lower than apo E 4/3 and LDLR AvaⅡ (+/+)individual’s. Conclusions The present study suggests that the LDLR polymorphism may be associated with interindividual variation in plasma cholesterol level. The effects of apo E and LDLR genes on cholesterol levels were independent from each other.