Abstract:Aim To observe the therapeutic effect of soybean isoflavones (SI) on the expression of peroxisome proliferator activated receptor α (PPARα), nuclear factor-kappa B(NF-κB), vascular cell adhesion molecule-1(VCAM-1), and investigate its mechanism of anti-atherosclerosis in metabolic syndrome rats. Methods Sixty male SD rats were randomly divided into normal diet group (n=10)and high-fat diet group (n=50)to induce metabolic syndrome model. Rats in high-fat diet group were fed with high-lipid high-salt high-sugar food for 20 weeks. After 20 weeks 37 metabolic syndrome rats were got successfully, and then these 37 metabolic syndrome rats were randomly divided into model group(n=9), fenofibrate-treated group(n=8), low dose (90 mg/kg) SI-treated group (n=10) and high dose (360 mg/kg) SI-treated group(n=10). Blood lipid level was measured after 4 weeks and mRNA level of PPARα of rat liver was determined using fluorescent quantitative PCR (FQ-PCR). The expressions of NF-κB and VCAM-1 of rat aorta were observed by immunohistochemistry. Results In high dose SI-treatment group, the low density lipoprotein cholesterol (LDLC),triglyceride (TG) and total cholesterol (TC) levels were significantly lower than those in model group(P<0.01), while the expression of PPARα was increased compared with model group. The expressions of NF-κB and VCAM-1 in model group were higher than those in control group. Compared with model group, high-dose SI decreased the expressions of NF-κB and VCAM-1 on the rat aorta. Conclusion SI can regulate the blood lipid level in metabolic syndrome rat, reduce the level of atherosclerosis factor, and can effectively stop the formation of atherosclerosis. Its mechanism may be associated with up-regulating the expression of PPARα mRNA by SI.

Abstract:Aim To investigate the mechanisms of chlamydia pneumoniae (Cpn)-induced human monocytic cell line (THP-1)-derived foam cell formation, the expression of scavenger receptor A (SR-A1) and CD36 were examined. Methods THP-1-derived macrophages were incubated with or without increasing concentrations of Cpn (1×105 to 1×106 IFU) for 0 to 72 h. Lipid droplets in cytoplasm were observed by oil red O staining. The contents of intracellular cholesterol ester were detected by enzyme-fluorescence. The expression of SR-A1 and CD36 at mRNA and protein levels were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western-Blot, respectively. Results Higher concentrations of Cpn infection (5×105 and 1×106 IFU) for 48 h result in the large accumulation of lipid droplets and the ratio of cholesteryl ester to total cholesterol was much higher than 50% in THP-1-derived macrophages when co-cultured with low density lipoprotein (LDL). Although Cpn infection had no effect on CD36 mRNA and protein expression, it up-regulated the expression of SR-A1 mRNA and protein in concentration-and time-dependent manner in THP-1 macrophages when co-cultured with LDL. Conclusions Cpn induces THP-1-derived foam cell formation by up-regulating the expression of SR-A1, which may provide a new evidence for the development and progression of atherosclerosis initiated by Cpn infection.

Abstract:Aim To investigate the effect of gemfibrozil on myocardium ischemia/reperfusion injury in hypercholesterolemic rabbits. Methods Twenty-four New Zealand white rabbits were divided into ischemia reperfusion (I/R) group, gemfibrozil treatment and ischemia reperfusion group, and sham operation group, which were fed with high cholesterol diet for 9 weeks to establish hypercholesterolmia rabbits model. And 200 mg/(kg·d) gemfibrozil was given for a week in gemfibrozil group on the nineth week. Acute myocardial ischemia reperfusion injury model was built through ligating the left anterior descending of coronary artery in rabbits. The serum lipid levels were measured in the different experiment stages. The ultrastructure of the myocardial cells by transmission electron microscope was observed and the sizes of infarct myocardium were detected in each group. The expression of peroxisome proliferator-activated receptor alpha (PPARα) and fatty acid translocase (CD36) mRNA were detected by reverse transcription polymerase chain reaction(RT-PCR). Results Rabbits fed with cholesterol-riched diet showed higher serum levels of total cholesterol(TC), low density lipoprotein cholesterol (LDLC) (P<0.05). Gemfibrozil did not change serum lipids levels during the feeding period. The ultrastructure of myocardium cell was slightly destroyed and the myocardial infarct size was significantly smaller in gemfibrozil treatment group than I/R group. The mRNA levels of PPARα and CD36 were decreased in I/R group compared with sham operation group, and there were no difference between gemfibrozil treatment group and sham operation group. Conclusion The short-term gemfibrozil treatment reduced the myocardial infarct size and up-regulted expression of PPARα and CD36 mRNA in myocardial after ischemia/reperfusion.

Abstract:Aim To investigate the effect of simvastatin on the proliferation and migration of vascular smooth muscle cells and the expression of vascular endothelial growth factor mRNA. Methods The atherosclerotic vessel injured model was established and male Sprague-Dawley rats were divided into four groups randomly: normal control group, atherosclerotic injured group, low dose simvastatin group and high dose simvastatin group. The rats were put to death after 4 weeks. The serum lipid and the ratio of intima/ (intima + media) of thoracic aorta and left common carotid artery were detected, the expression of vascular endothelial growth factor on blood vessel was determined by immunohistochemistory and RT-PCR method. Results Simvastatin didn’t show biphasic effect on the proliferation and migration of vascular smooth muscle cells. Low dose simvastatin didn’t promote the proliferation and migration of vascular smooth muscle cells, while high dose simvastatin showed inhibition effect on the proliferation and migration of vascular smooth muscle cells, which was independent on lipid decreased. Simvastatin could diminish the vascular endothelial growth factor mRNA expression of vascular smooth muscle cells. Moreover, high dose simvastatin could significantly decrease the vascular endothelial growth factor mRNA expression. Conclusions Simvastatin might have inhibitive effect on vascular smooth muscle cells by decreasing the expression of vascular endothelial growth factor in vascular smooth muscle cells.

Abstract:Aim To investigate the protective effect of Salvianolate on the injury of human umbilical vein endothelial cells (HUVEC) induced by high glucose in vitro, and to explore its underlying mechanism. Method HUVEC were respectively cultured with 5.5 mmol/L glucose(Glu)(control group), 30 mmol/L Glu (high glucose injury group), 30 mmol/L Glu+50 mg/L Salvianolate (low dose Salvianolate group), 30 mmol/L Glu+100 mg/L Salvianolate (medium dose Salvianolate group), 30 mmol/L Glu+ 200 mg/L Salvianolate (high dose Salvianolate group) for 48 h. The morphous of cell was observed by inverted phase contrast microscope, and the cell viability as well as the activity of glutathione peroxidase (GSH-Px), malondialdehyde (MDA) content, the secretary content of nitric oxide(NO)and endothelin-1(ET-1), in supernatant were measured. Results In high glucose injury group, low dose Salvianolate group, medium dose Salvianolate group and high dose Salvianolate group, the cell viability, activity of GSH-Px and MDA content, secrete of ET-1 and NO were significantly different compared with those in control group(P<0.01); In low dose Salvianolate group, medium dose Salvianolate group and high dose Salvianolate group, the cell viability, activity of GSH-Px and MDA content, secrete of ET-1 and NO were significantly different compared with those in high glucose injury group(P<0.01). Conclusion The Salvianolate can protect HUVEC induced by high glucose in vitro, and the mechanism may be associated with scavenging reactive oxygen radicals, reducing lipid peroxidation, and increasing the activities of intracellular antioxidase, inhibiting the secrete of ET-1.

Abstract:Aim To determine the role of Niemann-Pick C1-Like 1 (NPC1L1) in hepatic steatosis induced by liver X receptor (LXR) agonist in mice. Methods After being fed with 0.015% cholesterol diet for 21 days and gavaged with vehicle or T0901317 [25 mg/(kg·day)] for 7 days, both C57BL/6 mice and NPC1L1 knockout (NPC1L1-KO) mice were anaesthetized, livers were weighed and hepatic lipids were extracted and measured by enzymatic methods. Relative hepatic sterol regulatory element-binding protein-1c (SREBP-1c), fatty acid synthase (FAS) and stearoyl-CoA desaturase-1 (SCD-1) mRNA levels were analyzed by real-time quantitative PCR. Results After being gavaged with T0901317 for one week, the livers of C57BL/6 mice enlarged from 1.1±0.1 g to 2.8±0.3 g, and hepatic triglyceride content markedly increased from 34.2±18.1 mg/g to 232.2±67.9 mg/g, which was associated with the significantly enhanced hepatic mRNA levels of SREBP-1,FAS and SCD-1. However in NPC1L1-KO mice, liver weight only increased from 0.9±0.1 g to 1.5±0.1 g, and hepatic triglyceride content increased from 43.7±26.5 mg/g to 104.9±62.1 mg/g after treated with T0901317. Though hepatic mRNA levels of FAS and SCD-1 enhanced, the hepatic mRNA level of FAS in NPC1L1-KO mice treated with T0901317 was still 63% lower than that in C57BL/6 mice T0901317 treatment. Conclusion The down-regulation of hepatic SREBP-1 and FAS induced by NPC1L1 elimination attenuates the LXR agonist-dependent hepatic steatosis in mice.

Abstract:Aim To investigate the effects of atorvastatin on TNF-α induced endothelial lipase secreting by rat vascular smooth muscle cells. Methods The rat vascular smooth muscle cells were obtained from thoracic aortas and cultured by tissue explant method. Rat vascular smooth muscle cells were first incubated in serum-free medium with 50 μg/Ltumor necrotic factor-alpha; 1 hour later,atorvastatins with different concentration(1×10-7 mmol/L ,1×10-6mmol /L,1×10-5 mmol/L)were added to the medium. 24 hours later, cells of different groups were collected and endothelial lipase mRNA levels were determined by reverse transcription-polymerase chain reaction. In another group with atorvastatin concentration of 1×10-5 mmol/L, endothelial lipase levels were respectively determined after 6. 12 and 24 hours. Results Endothelial lipase expressions of atorvastatin intervent groups(1×10-7 mmol/L ,1×10-6 mmol /L,1×10-5 mmol/L) decreased significantly vs TNF-α group(P<0.05). The levels of endothelial lipase expression decreased initially with the atorvastatin concentration raise(P<0.05). Endothelial lipase expressions of atorvastatin time-intervent groups(6,12,24 h) decreased significantly vs. TNF-α group(P<0.05). The levels of endothelial lipase expression decreased initially with the time extended (P<0.05). Conclusions Atorvastatin can inhibit endothelial lipase mRNA secreting by tumor necrotic factor-alpha stimulated vascular smooth muscle cells, in a concentration-time dependent manner.

Abstract:Aim To explore the function of angiotensin Ⅱ (AngⅡ) in promoting osteopontin gene expression and protein synthsis of rat vascular adventitial fibroblasts. Methods Cultured cells were incubated with 1 different concentration of AngⅡ for different hours and AngⅡ plus AT1 receptor blocker,losartan or/and AT2 receptor blocker,PD123319. Then RT-PCR and WesternBlotting analysis were used to observe the effect of AngⅡ on osteopontin gene expression of rat vascular adventitial fibroblasts. Results AngⅡ(10-6 mol/L)obviously enhanced the expression of OPN mRNA in adventitial fibroblasts in time-dependent manner, the ratio of mRNA of osteopontin and β-actin in cells incubated with AngⅡ for 6,12 and 24 h increased by 65.43%, 97.03% and 105.11% respectively,as compared with that of the control (P<0.05), whereas there was no significant change in negative control at 24 h. AngⅡ could induce gene expression of osteopontin in dose-dependent manner, cells incubated with 10-8, 10-7, 10-6 and 10-5 mol/L AngⅡ for 12 h, there was up-regulation of OPN mRNA expression by 70.46%, 97.37%, 123.10% and 147.59% respectively (P<0.01).Pre-treatment with losartan(10-5 mol/L) and/or PD123319(10 mmol/L) for 1h in adventitial fibroblasts before AngⅡ(10-6 mol/L) used,the expression of OPN mRNA was inhibited by 58.9%, 0.92% and 59.7% respectively . Westernblotting analysis also showed that protein synthesis of osteopontin in cells incubated with 10-8, 10-7, 10-6 and 10-5 mol/L AngⅡ for 12h increased by 50.68%, 63.03%, 69.86% respectively ,as compared with that of the control(P<0.05). Pre-treatment with losartan(10-5 mol/L) but not PD123319 (10 mmol/L) before AngⅡ (10-6 mol/L) used, could inhibited the expression of OPN protein. Conclusions AngⅡ may up-regulate osteopontin gene expression and protein synthesis of rat vascular adventitial fibroblasts via angiotensin type 1 receptor.

Abstract:Aim To investigate the association between lipoprotein-associated phospholipase A2(Lp-PLA2) R92H single nucleotide polymorphisms (SNP) and susceptibility to coronary heart disease (CHD) and the degree of coronary artery stenosis. Methods 261 CHD patients and 263 normal controls were genotyped by using single-labeled probe technique. Results The frequencies of the RH genotypes and H allele were significantly higher in CHD patients than normal controls (P<0.05 and P<0.01). Compared with RR genotype, the relative risk for CHD in people with RH genotypes was 1.77 (P<0.01). The relative risk for CHD in people with H allele was 1.619 higher than R allele (P<0.01). Binamy logistic regression analysis showed that polymorphism of R92H was an independent risk factor for CHD. There was a significant increase of total cholesterol (TC) and low density lipoprotein cholesterol (LDLC) in the RH+HH genotype of CHD patients, while high density lipoprotein cholesterol (HDLC) was lower (all P<0.05). The ratio of multiple vessel diseases was significantly increased in the RH+HH genotype of CHD patients (P<0.05). Conclusions The polymorphism of R92H of Lp-PLA2 gene could be associated with risk of CHD in Han Chinese, persons with RH or HH genotypes take higher risk of suffering from CHD.

Abstract:Aim Through the confer of concentration of placenta growth factor(PLGF) and interleukin-10 (IL-10) to explore its early prediction and prognostic value in acute coronary syndrome (ACS) patients. Methods 90 patients were divided into experimental sub-STEMI group, NSTEMI/UAP group, SAP group and control group. The concentration of PLGF and IL-10 were measured by the method of enzyme linked immunosorbent assay (ELISA). The risk of follow-up cardiovascular events were detected during 30 days and 3 months, then the results were statisticly analyzed. Result The serum level of PLGF was higher in STEMI group and NSTEMI/UAP group than that of SAP group, but the concentration of IL-10 was lower in STEMI group and NSTEMI/UAP group than that of SAP group (P<0.01). The risk of cardiovascular events were increased in patients with higher serum level of PLGF, which was decreased in patients with higher serum level of IL-10 during 30 days and 3 months of follow-up. There was an inverse correlation between the serum level of PLGF and IL-10. Conclusions PLGF may be early indicators of ACS powerful, the balance between PLGF and IL-10 is a major determinant of outcome in patients with ACS.

Abstract:Aim To explore the relationship between the feature of carotid atherosclerotic plaque and cerebral hemodynamic status by 16-slice spiral computed tomography (CT). Methods 42 cases with carotid atherosclerotic plaques tested by carotid ultra-sonography had CT perfusion (CTP) and CT angiography (CTA). The CT values (housefield value,HV) of the major plaque components were assessed. According to different CT values, plaques were defined as stable (including lipid core and fibrous tissue) or instable (calcification ). Then CTP was used to evaluate the cerebral hemodynamic status. Results Among 42 cases, 25 cases had instable plaques. Of which a lipid core was found in 10 cases and fibrous tissue in 15 cases. Hypo-perfusion status was occurred in 14 cases, while 70% in lipid core, and 47% in fibrous tissue, but none in calcification tissue. Conclusion Instable plaque is more relative with hypo-perfusion than stable plaque.

Abstract:Aim To analyze the reason and the treatment of the complications of percutaneous coronary intervention (PCI). Methods From September 2002 to July 2006, PCI was performed in 566 patients, complications occured in 47 of them, which were retrospectively reviewed. The incidence rate of complications in anterior 283 patients was compared with that in post 283 patients , and the long-term incidence rate of the metal-naked stents was compared with that of rapamycin-coated stents. Results There were 6 no-reflow phenomenons and slow-reflow phenomenons (1.06%), 9 branch occlusion (1.59%), 4 coronary perforation (0.71%), 3 ventricular fibrillations (0.53%), 2 pseudoaneurysm (0.35%), 10 acute or subac thrombosis (1.77%), 1 long-term thrombosis (0.18%), 2 cerebral apoplexy (0.35%). Conclusion Great importance should be attached to precautions before operation. While the operation is on, it must be done properly and after it, the patients should be monitored closely.

Abstract:Aim To explore the association between coronary heart disease (CHD) and gene polymorphism of three key genes in renin-angiotensin system, including insertion/deletion (I/D) polymorphism of angiotensin converting enzyme (ACE) gene, M235T polymorphism of angiotensiongen (AGT) gene and A1166/C polymorphism of angiotensin Ⅱ type 1 receptor(AT1R) gene. Methods Single gene and gene linkage were respectively analyzed by polymerase chain reaction combined with restriction fragment length polymorphism (PCR-RFLP) in 110 patients with CHD and 80 healthy subjects. Results ① The frequency of DD genetype (43.6%) and D allele (60.5%) in CHD group were markedly higher than that in nomal control group (26.3% and 44.4% respectively, P<0.05). The frequency of TT genetype (66.4%) and T allele(78.6%) in CHD group were significantly higher than that in nomal control group (42.5% and 60.6% respectively, P<0.05). Both the genotype frequencies of AA and AC and the allele frequencies of A and C did not differ in CHD group and in nomal control group (P>0.05). ②The odds ratio (OR) estimated by combined analysis of those three genotypes was higher than either ACE-DD(3.395 vs 2.175) or AGT-TT (2.669) genotype estimated alone. Yet, OR decreased compared with ACE-DD and AGT-TT genotypes (6.098) estimated by combined analysis. Conclusions There is a strong correlation between CHD and I/D polymorphism of ACE gene and M235T polymorphism of AGT gene. No significant association between A1166/C polymorphism of AT1R gene and CHD is confirmed in this study. And combined analysis of the ACE-DD and AGT-TT genotype may enhance the predictability of CHD.

Abstract:Aim To observe the vascular structures anomalies in posterior circulation ischemia patients by digital subtractions angiography (DSA), and to compare incidences of the posterior circulation vascular abnormalities of transient ischemic attack (TIA) patients with that of the posterior circulation ischemic infarction patients. Methods 397 patients were selected from Nanjing Stroke Registry Program from January 2004 to May 2008, and diagnosed as posterior circulation ischemia that had digital subtraction angiography examination at the same time. Results Posterior circulation vascular abnormalities were detected in 304 patients and 435 lesions, and there was no difference between the TIA and posterior circulation infarction patients (P>0.05). In the 78 patients without risk factor, there were 25 patients who had vertebral artery hypoplasia or posterior circulation variations. Conclusion The incidence of posterior circulation vascular anomalies in posterior circulation ischemia patients is high; there is no difference between the TIA and posterior circulation infarction patients. In patients without risk factor, the incidence of vertebral artery hypoplasia or posterior circulation variation is high.

Abstract:Aim To explore the cardiovascular structure and function of ultrasonograph, blood-fat of healthy people by non-invasive ways, and analyze its action mechanism of arteriosclerosis. Methods 855 healthy people coming from 3 cities of Shenyang, Dalian and Beijing, were divided into 4 groups:≤44 years old group, 45~59 years old group, 60~74 years old group and ≥75 years old group. 55 items including life style, dietary structure, blood pressure, heart and carotid ultrasonograph, biochemistry, were measured to observe their changing with advancing age. Results 8 items including pulse pressure (PP), carotid artery intima-media thickness (IMT), carotid artery internal diameter (D), mitral valve anulus anterior wall A (MVAA), mitral valve anulus lateral wall E (MVLE), total cholesterol (TC), low density lipoprotein (LDL), high density lipoprotein (HDL) were correlated with aging significantly (r value was 0.521, 0.425, 0.258, 0.17, -0.25, 0.274, 0.181, -0.251 respectively with all P<0.01). Conclusions PP, IMT and carotid artery internal diameter are independent factors of the cardiovascular structure and function; IMT is the change of arteriosclerosis at earlier period; TC is the important risk factor of arteriosclerosis.