Qian Xiaoxian

The relationship of serum inflammatory factors, adiponectin and early atherosclerotic in type 2 diabetes

Objective To investigate the relationship of serum inflammatory factors, adiponectin and early atherosclerotic in type 2 diabetic patients (T2DM). Methods All 125 type 2 diabetic patients were divided into two groups according to the carotid intima-media thickness (CIMT) as follows: 65 cases of non-atherosclerotic diabetic patients with CIMT ≤0.9 mm and another 60 cases of atherosclerotic diabetic patients with CIMT > 0.9 mm or with plaque rapture being found. Thirty non-diabetic subjects were recruited as normal control. The clinical characteristics were collected for each patient. Serum fasting plasma glucose (FPG), glycation haemoglobin A1c (GHbA1c), fasting insulin (FINS), 2 h postprandic plasma glucose (2hPG), lipid profiles were measured. Serum levels of adiponectin, intercellular adhesion molecule−1 (ICAM-1), tumour necrosis factor-α (TNFα) and high sensitive C-reactive protein (hs-CRP) were also detected. Insulin resistance index (HOMA-IR) was calculated. Results (1) The serum levels of TNFα and hs-CRP, HOMA-IR were significantly higher in atherosclerotic patients than that of non-atherosclerotic and control subjects. However, the level of adiponectin in arteriosclerosis group was lower than other two groups. (2) There were negative correlation between adiponectin and CIMT, ICAM-1, TNFα, hs-CRP and HOMA-IR (r=0.574, −0.635, −0.681, −0.768, −0.752, p<0.01). There were negative correlation between HOMA-IR and ICAM-1, TNFα, hs-CRP (r=0.657, 0.706, 0.688, p<0.01). (3) TG, LDL-C and HOMA-IR were risk factors of T2DM complicated with arteriosclerosis according to multiple regression analysis. Conclusion The results suggested that inflammatory factors and adiponectin involved in the occurrence and development of diabetic vascular complications. The protective effects of adiponectin may exist in its anti-inflammation to improve insulin resistance and inhibit the development of atherosclerosis.

GW25-e4458 Effects of Ginsenoside Rb1 on vascular restenosis, SOD and MDA in rabbits with iliac artery injury

Objectives: we established a rabbit model of AF induced by atrial ischemia combined with rapid atrial pacing (RAP) to evaluate the relative contributions of electrical and histological remodeling to the inducibility and maintenance of AF. Methods: Twenty-four rabbits were randomly divided into an atrial ischemia combined with RAP group (Group I, n1⁄48), RAP group (Group P, n1⁄48) and an equal control group (Group C, n1⁄48.) An electrode sutured onto the left atrial appendage provided stimulation and recordings. Group I underwent RAP (1, 000 beats/minute) following successful ligation of the atrial branch of the right coronary artery. Group C do the pseudoperation without pacing, Group P underwent RAP (1, 000 beats/minute). Electrical and histological parameters were tested at three intervals: 1 hour, 1 week and 3 weeks. Results: The rabbits in group I showed a higher rate of AF induction, shortening of the atrial effective refractory period (AERP), loss of the normal rate adaptation and intra-atrial conduction delay (IACD), and prolongation of the P-wave interval. With prolonged ischemia and RAP, the IACD and persistence of AF increased. Shortening of the AERP and loss of the normal rate adaptation appeared at 1 hour and reached its maximum after 1 week. After 3 weeks, pathological examination of group I animals showed myocardial ischemia, edema, focal necrosis, and fibrosis, more evident in the right atrium. Group C showed no pathological changes. Conclusions: Atrial ischemia combined with RAP resulted in evident electrical and histological remodeling of the atrium, which effectively promoted the inducibility and maintenance of AF.

GW25-e4460 Diagnostic accuracy of 320-slice dynamic volume computed tomography angiography for detection of coronary artery stenosis compared with selective coronary angiography

(4.68 1.19; 5.10 2.34;5.17 1.38) and 7:00w9:00 (4.67 1.65; 4.90 2.50; 5.17 2.10)were lower significantly (P<0.05) when compared with those in controls (4.51 0.87; 3.21 1.23;5.30 1.89; 5.26 2.06), especially in MS1 (P<0.01); global CAS was significantly inversely correlated with stiffness parameter b (r1⁄4-0.333, P<0.01), PWVb (r1⁄4-0.358, P<0.01), positively correlated with AC (r1⁄40.333, P<0.01). (3) PCT and ADP was prolonged and ADT was shorter in patients with MS than in controls (P<0.01).PCT was positively correlated with b (r1⁄40.231, P<0.05), PWVb (r1⁄40.227, P<0.05), inversely correlated with global CAS (r 1⁄4 -0.29, P<0.01); ADT was positively correlated with global CAS (r 1⁄4 0.244, P<0.01). Conclusions: In patients with MS, Carotid artery stiffness increased, Circumferential strain decreased, especially in distal arteries wall, and CAS will further reduce with the progression; PCT and ADP was prolonged and ADT was shorter in patients with MS than in controls, indicating Cardiac contraction and pressure produces significantly delayed and vascular recoil shortened. Speckle tracking imaging can evaluate the change of carotid artery structure and function in patients with MS, and has important clinical value for early clinical detection and intervention treatment in process of MS.

GW24-e0762 Effects of Tongxinluo and Homocysteine on proliferation of human umbilical vein endothelial Cells

Objectives To investigate the effects of Tongxinluo and Homocysteine (Hcy) on proliferation of human umbilical vein endothelial cells (Huvecs) in vitro. Methods Isolation,culture and identification of Huvecs:With the informed consents of puerperants, the normal fetal umbilical cords were obtained through uterine-incision delivery in Third Affiliated Hospital, Sun Yat-sun University. Huvecs were isolated by Percoll density gradient centrifugation from fetal umbilical cords with digestion of collagenase type perfusion, and then suspended in Medium 199 supplemented with 20% FBS, 2 mM L-glutamine, 50 μg/ml endothelial cell growth factor, 100 μg/ml heparin and 100μg/ml streptomycin cultured in 0.05 volume fraction of CO2 incubator at 37°. The culture solution was changed after 24 hours, Huvecs cultured to 80% confluence, the passage was developed with 0.25% trypsinase and 0.02% EDTA. After the Huvecs were identified by flow cytometry with the cell marker CD34, The second or third passage was used for study. MTT assay was used to detect cell viability: Huvecs during the logarithmic phase were inoculated in 96-well plates with Medium 199 in 0.05 volume fraction of CO2 incubator at 37° for 48h, at 80% confluence, incubated in serum-free medium,and divided into two groups, eachgroup including eight parallel wells, blank control group was untreated, he latter cultured and added with Hcy of the concentrations of 0.1, 0.25, 0.5, and 1.0 mmol/L respectively,or Hcy 0.5 mmol/L+Tongxinluo of the concentrations of 0.1, 0.5, 1.0, and 2.0mg/ml. After the cells were cultured for 24 hr, 5 g/L MTT solution (20μL) was added, then dimethyl sulfoxide was used for vibration and dissolvement. Optical density of each hole was detected by using enzyme-linked immunosorbent assay to analyse different concentrations of Hcy and Tongxinluo on the proliferation of human mammary stemscells. Results Hcy of different doses inhibited the viability of Huvecs dose-dependently, the Optical density were 0.526 ± 0.017, 0.439 ± 0.010, 0.258 ± 0.008, and 0.209 ± 0.004 respectively, lower than that (0.578 ± 0.023) of blank control group (all P<0.01), accordingly,the inhibition rates of Hcy were 0.099 ± 0.079, 0.239 ± 0.046, 0.637 ± 0.007, and 0.553 ± 0.029 respectively (P<0.05, P<0.01); the growth rates of Hcy-induced Huvecs treatment by Tongxinluo of the concentrations of 0.1, 0.5, 1.0, and 2.0mg/ml were 0.682 ± 0.312,0.710 ± 0.040, 0.855 ± 0.130, 0.784 ± 0.031. Conclusions Hcy can inhibited the viability of Huvecs dose-dependently, and this effect is most prominent at the concentrations of 0.5 mmol/L for 24 hours; Tongxinluo can enhance the proliferation activity of Huvecs, and this effect is most prominent at 1.0mg/ml for 24 hours.

GW24-e2123 Effect of Sini Decoction on the expression of Sirt-1 and eNOS system in EAhy926 cells injured by homocysteine

Objectives To detect the effect of Sini Decoction on the expression of Sirt-1 and eNOS in EAhy926 cell injured by homocysteine. Methods Model of EAhy926 cell injured by homocysteine was made,the protection on the EAhy926 cell of Sini Decoction with different dosages were observed. NO concentration of cell culture fluid was detected by nitrate reductase method, effect of Sini Decoction on the expression of protein of Sirt-1 and eNOS in EAhy926 cell were observed by Western-blot, and effect of Sini Decoction on the expression of mRNA of Sirt-1 and eNOS in EAhy926 cell were observed by fluorescent quantitation PCR. Results After Model of EAhy926 cell injured by homocysteine was made, we found that cultured with 0.5,1.0,2.0,4.0,8.0 μmol/L homocysteine, cells grew less than cultured with normal culture medium, and with the increase of homocysteine concentration, the number of attached cell grew downwards obviously, as culturing with homocysteine 4.0 μmol/L for 24h did lower damage to cells and could induce effective cell injuring, it was made to be the model of injury. To detect the effect of Sini Decoction on EAhy926 cell injured by homocysteine, well growing EAhy926 cells were cultured in culture plate. 24 h later, cells were cultured with DMEM medium containing 2% fetal calf serum for 8 hours to make cells hungry, then cultured with medium containing Sini Decoction 0, 0.25, 0.5, 1.0g/ml respectively for 30 minutes,then cultured with medium containing homocysteine 4.0 μmol/L for 24 h. It was found that, compared with control group, attached cells in Sini Decoction groups grew better, and attached cells in Sini Decoction 1.0g/ml plus homocysteine 4.0 μmol/L group grew best. 4.0 μmol/L. Detected by nitrate reductase method, it was found that compared with control group, there was no obvious change of NO concentration of cell culture fluid in Sini Decoction 1.0 g/ml group, but in homocysteine 4.0 μmol/L medol group, NO concentration of cell culture fluid decreased oberviously, and in Sini Decoction groups, NO concentration of cell culture fluid increased, and in Sini Decoction 1.0g/ml plus homocysteine 4.0 μmol/L group it was the most obvious (p < 0.05). Detected by Western-blot, it was found that,campared with control group, there was no obervious change of protein of Sirt-1 and eNOS in Sini Decoction 1.0 g/ml group, but in homocysteine 4.0μmol/L model group, expression of Sirt-1 and eNOS protein weakened obviously, and in Sini Decoction groups, expression of Sirt-1 and eNOS protein enhanced, and in Sini Decoction 1.0g/ml plus homocysteine 4.0 μmol/L group it was the most obvious (p < 0.05). Detected by fluorescent quantitation, it was found that, compared with control group, there was no obvious change of mRNA of Sirt-1 and eNOS in Sini Decoction 1.0g/ml group, but in homocysteine 4.0 μmol/L model group, expression of Sirt-1 and eNOS mRNA weakened obviously, and in Sini Decoction groups, expression of Sirt-1 and eNOS mRNA enhanced, and in Sini Decoction 1.0 g/ml Plus homocysteine 4.0 μmol/L group, it was the most obvious (p < 0.05). Conclusions Homocysteine may injure EAhy926 cell by suppressing the expression of Sirt-1 then suppressing the expression of eNOS system, while Sini Decoction may protect EAhy926 cell by enhancing the expression of caveolin-1 then enhancing the expression of eNOS system.

GW24-e0410 Stress-induced premature senescence in primary human umbilical vein endothelial cells

Objectives Senescence is involved in many age-related diseases, such as atherosclerosis, hypertension and diabetes. So how to delay or solve the problems caused by ageing has become a hot topic in the past decade. To explore the best stress-induced premature senescence (SIPS) model in vitro, we applied different dosages of H2O2 to the primary human umbilical vein endothelial cells (HUVECs). Methods SIPS was induced by H2O2 with different concentration (0, 20, 40, 60, 80, 100 μM) in HUVECs. Senescence related gene was evaluated with western blot or real-time PCR. Senescence-associated beta-galactosidase (SA-β-gal) staining was used to examine the phenotype of senescence. Apoptosis was analysed by flow cytometry. Results The SA-β-gal staining result showed that the senescent cell was detected more significantly with H2O2≥40 μM compared with the control group. Also the mRNA of p21 was upregulated significantly with 40μM H2O2 and more. The PAI-1 protein was increased significantly with 60 μM H2O2 and more. In the apoptosis aspect, there was a significant increased number of apoptotic cells in 80 and 100 μM group. Conclusions H2O2 can induce senescence of the primary HUVECs. In consideration of the its apoptotic effect, the 60 μM H2O2 could be the best concentration to induce senescence. Because of the high relevance with human, the primary HUVECs is a better candidate applied to explore the mechanism and the improved treatment of senescence in vitro. Besides H2O2, there are some other ways to induce senescence, such as UV and gene knock down. Among these methods, H2O2 induced senescence via oxidative stress is the most relavent to the enviroment in the aged population.

EFFECTS OF JIAWEI BUYANG HUANWU DECOCTION ON VASCULAR STENOSIS AND TGF-β1 AFTER BALLOON INJURY OF RABBIT ILIAC ARTERY

Objectives To investigate the effects of Jiawei Buyang Huanwu Decoction on vascular stenosis and transforming growth factor -β1 (TGF-β1) after iliac artery were injured by balloon in diet-induced atherosclerotic rabbits. Methods 24 male New Zealand albino rabbits were equally randomized into control group, model group and drug group. The iliac arteries of the rabbits in the latter two groups were subjected to balloon injury. Four weeks later, serum TGF-β1 level was assayed, Endothelial hyperplasia, eNOS Protein and mRNA expression were observed in injured iliac artery. Results Optical microscope revealed narrowed vascular lumen, thicken intima and numerous arteriosclerotic plaques in the model group compared with the control group, whereas the vascular lumen and intima thickness remained basically normal in drug group. The serum TGF-β1 level was lower in drug group than that of model group, Immunohistochemistry and RT-PCR results showed that eNOS protein and mRNA expression was lower in rabbit iliac artery of drug group than that in model group. Conclusions Jiawei Buyang Huanwu Decoction can lessen intimal hyperplasia and vascular stenosis in iliac artery injury rabbits, and the mechanism of which may be related to decrease in TGF-β1 protein and gene expression.

THE EFFECT OF JIAWEI BUYANG HUANWU DECOCTION ON THE NO SYSTEM AFTER ILIAC ARTERY BALLOON INJURED IN RABBITS

Objectives To investigate the effect of Jiawei Buyang Huanwu Decoction in preventing vascular restenosis and on the nitric oxide (NO) system after rabbit iliac artery balloon injury. Methods 24 male New Zealand albino rabbits were equally randomised into control group, model group and drug group. The iliac arteries of the rabbits in the latter two groups were subjected to balloon injury. Four weeks later, serum NO synthases (NOS) activity and endothelial nitric oxide synthase (eNOS) level was assayed, Endothelial hyperplasia, eNOS Protein and mRNA expression were observed in injured iliac artery. Results Optical microscope revealed narrowed vascular lumen, thicken intima and numerous arteriosclerotic plaques in the model group compared with the control group, whereas the vascular lumen and intima thickness remained basically normal in drug group. The serum NO level and total NOS activity were lower in model group than that of control group and drug group, while there is no difference between the latter two. Immunohistochemistry and RT-PCR results showed that eNOS protein and mRNA expression was stronger in rabbit iliac artery of control group and drug group than that in model group. Conclusions Jiawei BHD can lessen intimal hyperplasia and vascular stenosis in iliac artery injury rabbits, this effect is possibly related with that Jiawei BHD activated the NO system.

THE CHANGE OF SERUM LEVEL OF PIGMENT EPITHELIUM-DERIVED FACTOR IN CORONARY HEART DISEASE

Objectives To investigate the change of serum level of Pigment epithelium-derived factor (PEDF) in coronary heart disease patients. Methods 40 coronary heart disease patients, 29 males and 11 females were recruited into this study, which contains acute myocardial infarction 6 cases, unstable angina pectoris 15 cases, stable angina 19 cases, according to the New York Heart Association (NYHA) diagnostic criteria, ruling out infectious diseases, tumour, renal inadequacy, diabetes, Hepatic function damage, cardiac inadequacy, hyperthyrea, anaemia and hyperthermy. Control group contains 35 healthy people with similar gender, age and social background. Venous blood on an empty stomach were drawn next day after admission and were put in tubes, and were centrifuge in 1000 rpm for 10 min, then blood serum were imbibed and stored under −80°C. The serum of normal control group were taken in similar ways. Serum level of PEDF and IL-6 were determined by ELISA Kits (Bought from USCN life science Company, Wuhan, China and KeyGEN Biotech Company, Nanjing China). Meanwhile, serum level of total cholesterol, triglyeride and lipoprotein cholesterol were determined. Degree of stegnosis of coronary by Gensini scoring system based on the result of coronary arteriongraphy. Results The serum level of PEDF (8.39–2.64 ug/ ml) and Il-6 (7.40–4.41 pg/ml) in coronary heart disease group were higher than PEDF (6.952.92 µg/ml) and Il-6 (4.09–3.70 pg/ml) in control group (p<0.05). There was no deference between two groups in serum level of total cholesterol, triglyeride and lipoprotein cholesterol. There was no correlativity between serum level of PEDF and Gensini scores by Pearson correlation analysis (r=−0.166, p>0.05), but there was correlativity between serum level of PEDF and IL-6 (r=0.241, p<0.05). Conclusions The results suggest that Pigment epithelium-derived factor increases greatly in CHD patients, Pigment epithelium-derived factor may be an important factor of atherosclerosis.

EFFECT OF SINI DECOCTION ON THE EXPRESSION OF CAVEOLIN-1 AND ENOS IN EAHY926 CELL INJURED BY HOMOCYSTEINE

Objectives To detect the effect of Sini Decoction on the expression of Caveolin-1 and eNOS in EAhy926 cell injured by homocysteine. Methods Model of EAhy926 cell injured by homocysteine was made, the protection on the EAhy926 cell of Sini Decoction with different dosages and different durations were observed. The effection of Sini Decoction on the expression of protein of Caveolin-1 and eNOS in EAhy926 cell were observed by Western-blot, and effection of Sini Decoction on the expression of mRNA of Caveolin-1 and eNOS in EAhy926 cell were observed by fluorescent quantitation PCR. Results After model of EAhy926 cell injured by homocysteine was made, we found that cultured with 0.5, 1.0, 2.0, 4.0, 8.0 µmol/l homocysteine, cells grew less than cultured with normal culture medium, and with the increase of homocysteine concentration, the number of attached cell grew downwards obviously, as culturing with homocysteine 4.0 µmol/l for 24 h did lower damage to cells and could induce effective cell injuring, it was made to be the model of injury. To detect the effection of Sini Decoction on EAhy926 cell injured by homocysteine, well growing EAhy926 cells were cultured in culture plate. 24 h later, cells were cultured with DMEM medium containing 2% fetal calf serum for 8 h to make cells hungry, then cultured with medium containing Sini Decoction 0, 0.25, 0.5, 1.0 g/ml respectively for 30 min, then cultured with medium containing homocysteine 4.0 µmol/l for 24 h. It was found that, compared with control group, attached cells in Sini Decoction groups grew better, and attached cells in Sini Decoction 1.0 g/ml plus homocysteine 4.0 µmol/l group grew best. Detected by western-blot, it was found that, compared with control group, there was no obvious change of protein of Caveolin-1 and eNOS in Sini Decoction 1.0 g/ml group, but in homocysteine 4.0 µmol/l medol group, expression of Caveolin-1 protein enhanced obviously, expression of eNOS protein weakened obviously, and in Sini Decoction groups, expression of Caveolin-1 protein weakened, and expression of eNOS protein enhanced, and in Sini Decoction 1.0 g/ml plus homocysteine 4.0 µmol/l group it was the most obvious p<0.05). Detected by fluorescent quantitation, it was found that, compared with control group, there was no obvious change of mRNA of Caveolin-1 and eNOS in Sini Decoction 1.0 g/ml group, but in homocysteine 4.0 µmol/l medol group, expression of Caveolin-1 mRNA enhanced obviously, expression of eNOS mRNA weakened obviously, and in Sini Decoction groups, expression of Caveolin-1 mRNA weakened, and expression of eNOS mRNA enhanced, and in Sini Decoction 1.0 g/ml Plus homocysteine 4.0 µmol/l group, it was the most obvious p<0.05). Conclusions Homocysteine may injure EAhy926 cell by enhancing the expression of caveolin-1 then suppressing the expression of eNOS, while Sini Decoction may protect EAhy926 cell by suppressing the expression of caveolin-1 then enhancing the expression of eNOS.