Abstract:AimTo observe the changes of human umbilical vein endothelial cells (HUVEC) in function and membrane surface ultrastructure during the lipid peroxidation.MethodsHUVEC were induced by 100 mg/L oxidized low density lipoprotein (ox-LDL) as experiment group, then incubated for 0 h, 4 h, 8 h and 16 h.HUVEC in the PBS as control group was processed likewise.Cell viability was measured by MTT assay.The functional status of HUVEC was determined by detecting content of nitric oxide (NO) in the cultured cell supernate with nitric acid reduction assay.Atomic force microscope (AFM) was used for observation to membrane surface ultrastructure of HUVEC.ResultsThe research showed that the proliferation ability of HUVEC in the experimental group were inhibited, cell functions were attenuated, and the longer incubation, the more significantly effects.At the same time, the eminentias on cell surface became larger, distribution irregularity, even some caveolaes and holes appeared.However, there was no change in control groups.Cell surface roughness analysis showed that roughness on 0 h, 4 h, 8 h and 16 h experimental groups were 13.666±2.196 nm, 15.904±2.203 nm, 17.688±2.076 nm and 21.609±1.867 nm, respectively.There were significant differences between each two (p<0.05).In the comparison, the control group were 13.627±2.218 nm, 13.659±2.183 nm, 13.665±2.175 nm and 13.974±2.478 nm, respectively, and no significant differences between each two (p>0.05).Compared with the control group, roughness in the experimental groups after 4 hours induction was significantly higher (p<0.05).ConclusionsIt was demonstrated that the cell function of endothelial cells were weakened and membrane surface ultrastructure changed in an early stage of lipid peroxidation, and perform a time-dependent rule.

Abstract:Aim To study the effect and possible mechanism of mangiferin on myocardial ischemia reperfusion (MIR) injury in rats. Methods Fouty-eight SD rats were randomly divided into six groups: sham group, model group, mangiferin groups of 10 mg/kg, 20 mg/kg, 40 mg/kg and Diaoxinxuekang group. Each group were injected with corresponding concentrations of mangferin, Diaoxin xuekang or equal volume of saline gastrogavaged for 21 d. 0ne hour after the last administration myocardial ischemia reperfusion models were obtained by ligated left anterior descending coronary artery 40 minutes and followed by 120 minutes reperfusion and sham group was given all the procedures except ligation. Ultrastructure of myocardium with TEM and infiltration of polymorpho nuclear neutrophils (PMN) in myocardium with myeloperoxidase (MPO) were observed; serum activity of lactate dehyd -rogease (LDH), myocardial activity of superox -ide dismutase (SOD) and contents of malondialdehyde (MDA) were detected respectively. Results Mangiferin could improve myocardial pathology and ultrastructurein mangiferin 10 mg/kg group the activities of SOD was higher (P<0.05); the contents of MPO, MDA, LDH were significantly lower (P<0.05). Conclusion Mangiferin can protect MIR. The myocardial protective mechanism of it may be realized by enhancing the activation of SOD activity,enhancing myocardial antioxygen capability, stabilizing myocardial cellular membrance and alleviating infiltration of polymerrphonuclear neutrophils(PMN) in myocardium.

Abstract:Aim To investigate the effects of 10-23DNA enzyme(ED5) on endothelial function and ultrastructure change of Carotid Artery after injured artery in rats. Methods Ninety-six rats were randomly divided into sham-group,control-group 1,control-group 2 and ED5-group(n=24).The models of endothelial denudation of rats were made by using balloon catheter,FITC-ED5 was delivered to the artery wall of ED5-group in a total of 200 μL solution.Rats of control-group1 were given 200 μL 1 mmol/L MgCl2.Rats of control-group2 were given FuGENE6 Reagent.Six rats were killed at the 3rd,7th,14th,21th day after balloon injury.The serum level of nitric oxide and nitric oxide synthase and endothelin were detected.At the same time,to observe pathological change of injured arteries by optical microscope and electron microscope. Results The serum level of nitric oxide and nitric oxide synthase in ED5-group were higher than those in 2 control groups,increased significantly on the 14th day(nitric oxide: 57.1±1.9 μmol/L vs 38.7±1.9 μmol/L and 57.1±1.9 μmol/L vs 38.3±1.9 μmol/L,p<0.05;nitric oxide synthase: 11.1±0.4 μmol/L vs 8.1±0.4 μmol/L and 11.1±0.4 μmol/L vs 8.0±0.4 μmol/L,p<0.05);but endothelin in ED5-group was lower than that in 2 control groups,obviously different on the 14th day(111.2±7.2 pg/L vs 136.6±7.2 pg/L and 111.2±7.2 pg/L vs 135.5±7.2 pg/L,p<0.05).Compared with ED5-group,the neointimal thickness in 2 control groups increased significantly on the 21th day after balloon injury(64.0±4.2 μm vs 81.1±4.9 μm and 64.0±4.2 μm vs 79.5±3.9 μm, p<0.01).The vascular smooth muscle cell of neointima in 2 control groups may be contractile type,but vascular smooth muscle cell of neointima in ED5-group may be synthetic type. Conclusion The ED5 can improve the endothelial function after artery injury and inhibit the phenotype transform of vascular smooth muscle cells,and reduce the degree of neointimal hyperplasia.

Abstract:Aim To investigate effect of cigarette smoking onmorphology and expression of intercellular adhesion molecule-1 (ICAM-1), ICAM-1 mRNA in vascular endothelial cells of rats brains and to further explore influence of cigarette smoking on atherogenesis. Methods 75 male Wistar rats were randomly subgrouped into long-term massive, short-term massive, long-term trifle, anti-smoking and normal control group. Immunohistochemistry technique, in situ hybridization technique and pathologic image analyzing system were used to determine the expression of ICAM-1, ICAM-1 mRNA in rats cerebral vascular endothelial cells. Results In contrast to normal control group, expression of ICAM-1, ICAM-1 mRNA in rats cerebral vascular endothelial cells increased in smoking (103.7±3.9 vs 99.7±7.1 vs 92.6±5.2 vs 94.8±5.1 vs 84.2±4.7 for the mean gray value of ICAM-1mRNA staining in normal control, anti-smoking, short-term massive, long-term trifle, long-term massive group, repectively; p<0.05), and ICAM-1, ICAM-1 mRNA were relevant to the duration and dose of smoking (p<0.05). Expression of ICAM-1 and ICAM-1 mRNA decreased after anti-smoking, and there was no significant difference compared with normal control group (p>0.05). The structure of endothelial cells changed greatly in smoking, and partly recovered after anti-smoking. Conclusion The upregulation of expression of ICAM-1 and ICAM-1 mRNA might be one of the important molecular mechanisms during the process of atherosclerosis in cigarette smoking rats.

Abstract:Aim To study the effects of stichopus variegates compound on the iliac arterial endothelial function and ultrastructure of rabbits after transluminal balloon angioplasty (TBA) of iliac atherosclerosis model. Methods 50 purebred New Zealand rabbits were randomly divided into four groups: Stichopus,Simvastatin,model and normal group. Endothelium of iliac arteries in the front three groups were denuded by balloon catheter and fed with 2% cholesterol,3% lard and 3% yolk mixed forage for six weeks,and then,atherosclerotic stenosis was showed by iliac angiography. Stichopus group [0.5 g/(kg·d)] and Simvastatin group [0.5 mg/(kg·d)] were fed by gastric canal in mouth cavity. At the same time NaCl solution (0.9%) were fed in model group. All the rabbits were fed in different cages and not limited drinking. After four weeks since TBA,blood was collected from left carotid artery to measure the changes of concentrations of endothelin (ET) and nitric oxide(NO). Results ①After four weeks since TBA,the concentrations of plasm ET in model groups were higher than the one in the Simvastatin and Stichopus groups,and the concentrations of serum NO decreased obviously; the concentrations of plasm ET in Stichopus groups had an increasing tendency compared with Simvastatin and normal groups. ②After four weeks since TBA,neointimal mean thickness of segment operated TBA in model groups increased significantly,compared with Simvastatin and Stichopus groups. There were greater extent of lumen stenosis in model groups than the one in Simvastatin and Stichopus groups. ③Compared with model groups there were small vessel smooth muscle cell body and nucleus,less organelles and more myofilament contents,better cell differentiation,and less lipid droplet were phagocyted in Simvastatin and Stichopus groups under the transmission electron microscope. ④There were less platelets adherence in Simvastatin and Stichopus groups under scanning electron microscope. Conclusion Stichopus variegatus compound can significantly inhibit vessel smooth muscle cells proliferation,lessen neointimal thickness,enlarge luminal areas,decrease extent of lumen stenosis and protect endothelial function after iliac arterial transluminal angioplasty of rabbits.

Abstract:Aim To study the structural changes of basilar arteries during hypertension and the effects of captopril. Methods The animal model of stroke prone renovascular hypertensive rats might be divided into two groups, hypertension group and captopril treated group; one sham operated group was used asnormotensive control group. Each group took the observation of the structure of the basilar arteries under light microscope and transmission electrical microscope as well as analyses of morphometry at 4, 8, 12 weeks respectively. Results There were no obvious microscopic changes in basilar arteries of the hypertension group postoperation 4 weeks. The media thickness and wall to lumen ratio of the hypertension group were higher than those of captopril and sham operated groups (p<0.05). The intercellular spaces are broadened slightly at 4 weeks after operation; there were moderate ultrastructural damages at the end of 8 weeks, with the organelle oedema and partial lysis, interstitial dropsy; there were obvious ultrastructural damages at the end of 12 weeks, such as smooth muscle cell myofilament degeneration, fragmentation, lysis, and endoplasmic reticulumnecrosis dilataltion, mitochondria vacuolation, smooth muscle cell necrosis. The ultrastructure appearances of the captopril treated group were nearly normal. Conclusions Hypertension can cause hypertrophy and ultrastructural damage of basilar arteries. Captopril can prevent media hypertrophy of basilar arteries in experimental hypertensive rats and protect the ultrastructure against injury factors.

Abstract:Aim: To study the morphological and quatitative change of elastic fiber in different stages of atherosclerotic lesions.Methods: The normal intima, fatty streaks and fibrous plaques, from 17 abdominal aortae of young men of nom-cardiovascular sudden death were observed with electrolic microscope.Results: The elastlc flber in normal intima appeared to be cord-like, while the e1astic fiber in the fatty streaks and fibrous plaques appeared to be patch-like.The content of elastic fiber was highest ln the fatty streaks, the lowest in the fibrous plaques. The elastic fiber content of normal intima was between the fatty streaks and fibrous plaques. The differences of elastic fiber content among three groups were statistically significant (P<0. 01 ).Conclusion: With the developement of atherosclerosls, the elastlc fiber has not only morphological changes but also content change. This indicate that the elastic fiber plays a very irnportant role in the formation and developement of atherosclerosis.