Abstract:Aim To investigate the effect and possible mechanism of total flavones of dracocephalum moldavica (TFDM) on atherosclerosis lesions in apolipoprotein E gene deficient (ApoE－/－) mice. Methods ApoE－/－ mice were randomly divided into model group, simvastatin group and TFDM high, medium, low dose group, C57BL/6J mice were set as the normal control group. After treated for 12 weeks, the mice were sacrificed and plasma lipids were determined, the area of atherosclerotic plaques and the ratio of plaque area to aorta were measured by HE staining, the expressions of intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), proliferating cell nuclear antigen (PCNA) in aortic atherosclerotic lesions were detected by immunohistochemistry analysis. Results Compared with the model group, the levels of triglyceride (TG) and low density lipoprotein cholesterol (LDLC) was decreased significantly in treatment group. The atherosclerotic lesions reduced and the ratio of plaque area to aorta decreased in TFDM groups. The expression of ICAM-1, VCAM-1 and PCNA were down-regulated by TFDM. Conclusion TFDM can inhibit atherosclerosis formation which involved in decreasing the level of plasma lipids and weakening the expression of ICAM-1, VCAM-1 and PCNA in the aorta wall.

Abstract:Aim To investigate the relationships between intercellular adhesion molecule-1 （ICAM-1）, vascular cell adhesion molecule-1 （VCAM-1）, and intraplaque angiogenesis. Methods Twenty-five New Zealand white rabbits underwent balloon-induced abdominal aortic wall injury and were fed with a diet of 1% cholesterol to establish a rabbit model of atherosclerosis. In week 4, 6, 8, 10 and 12 after balloon injury every 5 rabbits were euthanasia respectively. Tissue samples were taken from the abdominal aorta. Some segments were embedded in paraffin and cut into 5 μm thick sections for staining with H&E and reacted with platelet-endothelial cell adhesion molecule-1 （PECAM-1, i.e. CD31）, rat anti-rabbit macrophage antibody-11 （RAM-11）, ICAM-1, and VCAM-1. Results From week 4 to week 12, the expression of ICAM-1 and VCAM-1 increased with the development of atherosclerotic plaque in the abdominal aorta of rabbits.

Abstract:AimTo study the effect and mechanism of rhynchophylla total alkaloids, sinapine cyanide sulfonate and its component compatibility on change of vascular endothelial cells induced by the tumor necrosis factor-α(TNF-α).MethodsRat vascular endothelial cells were cultured in vitro, and vascular endothelial cell injury model was established by TNF-α.Then changes of cell morphology were observed before and after treatment; intercellular adhesion molecule-1(ICAM-1), vascular cell adhesion molecule-1(VCAM-1) and rat endothelin-1 (ET-1) were determined by Elisa method, mRNA of ICAM-1, VCAM-1 and ET-1 by RT-PCR method and the level change of endothelium-derived relaxing factor (NO) by nitrate reductase method.ResultsThe vascular endothelial cell injury model could be established in vitro successfully by TNF-α (5 μg/L).Compared with TNF-α group, treated groups could improve vascular endothelial cell morphology, lower the secretion of ICAM-1,VCAM-1 and ET-1.Compared with rhynchophylla total alkaloids group and sinapine cyanide sulfonate group, the component compatibility group was better on heightening the secretion of NO, down-regulating the expression of ICAM-1 mRNA(P<0.05).On down-regulating the expression of mRNA of VCAM-1

Abstract:AimTo observe the effect of glucagon-like peptide-1 (GLP-1) receptor agonists exenatide on nuclear factor-κB (NF-κB) activity as well as the expression of its downstream inflammatory cytokins such as intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) induced by high glucose in human umbilical vein endothelial cells (HUVEC) and to explore mechanism of protective effect of exenatide on vascular complication of diabetes.MethodsHUVEC were cultured in vitro and incubated under serum-free conditions with 25 mmol/L glucose for 48 hours.Exenatide at different concentrations (10－8 mol/L, 10－7 mol/L and 10－6 mol/L) was added concurrently with glucose stimulation.Meanwhile ICAM-1 and VCAM-1 in the cell culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA).The expression of NF-κB p65, ICAM-1 and VCAM-1 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR).Results25 mmol/L D-glucose significantly increased NF-κB p65 mRNA expression as well as the supernatant content and mRNA expression of ICAM-1 and VCAM-1 in HUVEC (p<0.01)， which was inhibited by different concentrations of exenatide (p<0.01)， in a dose-dependent manner.ConclusionsGLP-1 receptor agonist exenatide could improve endothelial dysfunction via inhibition of NF-κB activity and its downstream inflammatory response in vascular endothelial cells induced by high glucose in a dose-dependent manner, which could prevent and improve vascular complication of diabetes.

Abstract:Aim To investigate the protective effect of Astragali radix extract on vascular cell adhesion molecule-1 expression of mice vascular endothelial cell against tumor necrosis factor-α(TNF-α). Methods Adhesion model was established by THP-1 cells and mice endothelial cell in vitro.The cells were pretreated by different dose and different time of Astragali radix extract before induced by TNF-α,and the adhesion rate were detected.The levels of vascular endothelial cell adhesion molecule VCAM-1 in the cell culture were determined with ELISA.The expression of VCAM-1 and NF-κB subunit(p65) were evaluated by Western blot. Results The expression of VCAM-1 and NF-κB was increased obviously after induced by TNF-α;While the expression of VCAM-1 and the effect of NF-κB p65 protein nuclear translocation induced by TNF-α were inhibited after pretreatment of Astragali radix extract in a dose-and time-dependent manner.The reduction of adhension of monocytes to endothelial cells,the down-regulation of the expression of VCAM-1 and reduc-tion of the expression of NF-κB were apparent(P<0.05) at the concentration of 120 mg/L preincubated 4～8 h. Conclusion Astragali radix extract can inhibit the TNF-α-induced expression of VCAM-1 and reduce the adhension of monocytes,by which the damage to vascular endothelial cells was relieved.The mechanism may be related to the role of inhibiting the activation of NF-κB.

Abstract:Aim To investigate the effects of advanced glycation end products(AGE) on the expression of monocyte chemoattractant protein-1(MCP-1) and vascular cell adhesion molecule-1(VCAM-1) in human umbilical vein endothelial cell(HUVEC)and the intervention effect of atorvastatin. Methods Collagenase was used to isolate the endothelial cell from human umbilical vein;HUVEC were identified by immunocellochemistry and morphology;RT-PCR was performed to detect MCP-1,VCAM-1 and recepter for AGE(RAGE) mRNA expression;Reactive oxygen species(ROS) detectionkit was used to examine the level of ROS in HUVEC and inversion fluoescence microscope was used to observe the ROS level. Results The cultured cells were oval or polygon and retained cobblestone appearance through inverted phase contrast microscope,brown particles could be observed in cytoplasm(CD31 or CD34 positive cells) after immunocytochemical staining with CD31 or CD34;In comparison with control group,AGE(10-4~10-1 g/L)promoted MCP-1 and VCAM-1 mRNA expression in a concentration-dependent manner in HUVEC.The expression of MCP-1 and VCAM-1 mRNA was significantly elevated by AGE(10-4 g/L) compared with the control group(0.26±0.02 vs 0.17±0.04;0.22±0.02 vs 0.08±0.01,P<0.01);Compared with AGE group,atorvastatin(0.1,1,10 μmol/L) diminished MCP-1 and VCAM-1 mRNA expression induced by AGE in HUVEC in a concentration-dependent manner;Atorvastatin in a dose of 1 μmol/L,could significantly decrease the expression of MCP-1 and VCAM-1 mRNA induced by AGE(0.63±0.11 vs 1.03±0.07;0.21±0.03 vs 0.83±0.10,P<0.01);The level of ROS in AGE group was higher than that in control group,atorvastatin could obviously decline the ROS level induced by AGE in HUVEC.Furthermore,atorvastatin(0.1 μmol/L) was able to decrease RAGE mRNA expression remarkly induced by AGE in HUVEC(0.63±0.05 vs 1.19±0.12,P<0.01). Conclusion AGE could significantly increase MCP-1 and VCAM-1 mRNA expression in HUVEC;Atorvastatin could decrease the oxidative stress and inflammation gene expression induced by AGE in HUVEC through inhibiting the expression of RAGE.

Abstract:Aim To study the non-lipid mechanisms of atorvastatin treating atherosclerosis by observing the changes of oxidative stress and inflammatory response. Methods The rabbit model of atherosclerosis were prepared by feeding the high fat diet and immune stimulation,then 0.435 mg/(kg·d) suspension of atorvastatin was given for 1 month.It was observed for the rabbit serum superoxide dismutase(SOD) activity,malondialdehyde(MDA) and oxidized low density lipoprotein(ox-LDL) levels and lectin-like oxidized low density lipoprotein cholesterol receptor-1(LOX-1),vascular cell adhesion molecule-1(VCAM-1),intercellular adhesion molecule-1(ICAM-1),monocyte chemoattractant protein-1(MCP-1) gene and protein expression. Results Compared with the control group,rabbit serum SOD activity was significantly decreased(P<0.01),serum MDA,ox-LDL was significantly higher(P<0.01);mRNA and protein expression of aortic LOX-1,VCAM-1,ICAM-1,and MCP-1 were significantly increased(P<0.05 or P<0.01) in atherosclerosis model group.Compared with model group,rabbits serum SOD activity was significantly increased(P<0.01),serum MDA,ox-LDL levels were significantly decreased(P<0.01);mRNA and protein expression of aortic LOX-1,VCAM-1,ICAM-1,and MCP-1 were significantly lower in atorvastatin group(P<0.05 or P<0.01). Conclusions Atorvastatin possesses anti-oxidative stress and inflammatory response,thereby it can protect the vascular endothelium and play a therapeutic role in atherosclerosis.

Abstract:AimTo investigate the effects of advanced glycation end products(AGE) and homocystein（Hcy）on the expression of vascular cell adhesion molecule-1(VCAM-1) mRNA via oxidative stress mechanism in human umbilical vein endothelial cell（HUVEC）.MethodsCollagenase was used to isolate the endothelial cell from human umbilical vein.RT-PCR was used to examine the expression of VCAM-1 mRNA.Inversion fluoescence microscope was used to observe the level of oxygen free radical.Results The expression of VCAM-1 mRNA was induced concentration-dependently by AGE(10－4～10－1 g/L) and Hcy(1.35×10－3～1.35 g/L); AGE combined with Hcy group could promote the expression of VCAM-1 mRNA respectively 7.78 times and 6.05 times in comparison with AGE group and Hcy group（1.09±0.18 vs 0.14±0.07；1.09±0.18 vs 0.18±0.06， P<0.01）; By comparison with AGE combined with Hcy group, the VCAM-1 expression of DPI group was obviously reduced(0.20±0.09 vs 1.19±0.23,P<0.01); In comparison with control group, AGE group and Hcy group could enhance oxygen free radical in HUVEC; The level of oxygen free radical in AGE combined with Hcy group was higher than that in AGE group and Hcy group; DPI could significantly inhibit the level of oxygen free radical.ConclusionAGE and Hcy can enhance the level of oxygen free radical in HUVEC and increase the VCAM-1 expression, and they have the cooperative effect; The enhancement of oxygen free radical is an important factor on the expression of VCAM-1 mRNA; The above process is realized possibly through activating the NADPH oxidase.

Abstract:Aim To study the effects of vascular adventitial vascular cell adhesion molecule-1(VCAM-1) and intercellular adhesion molecule-1(ICAM-1) on atherosclerotic lesion formation. Methods Apolipoprotein E Knockout(ApoE-/-) mice and wild-type C57BL/6 black mice of 6 weeks were used in this experiment.All animals were fed hyperlipidic diet for 2,4 and 8 weeks.The ascending aorta was removed with the heart attached for serial sectioning.Some sections were stained by Movat method in order to observe the morphological changes of the tissues and measure the thickness of vascular adventitia.Some sections were stained with immunohistochemistry to observe the changes in the expression of vascular adventitial ICAM-1 and VCAM-1 at different stages. Results ApoE-/-mice of 6 weeks and C57BL/6 mice at any time point did not show any changes of adventitial thickness,nor expression of VCAM-1,nor intimal lesion formation.After hyperlipidic diet for 2 weeks in ApoE-/-mice,the adventitial thickness increased and the expression of VCAM-1 was found in the adventitia when there was no visible intimal lesion formation.After hyperlipidic diet for 4 weeks the foam cells were showed in the intima and the atherosclerotic lesion developed after hyperlipidic diet for 8 weeks in ApoE-/-mice.The adventitial thickness and the expression of VCAM-1 in adventitia and intimal lesion of ApoE-/-mice increased.The expression of ICAM-1 in the adventitia and the intima of ApoE-/-mice increased gradually with increasing duration of the hyperlipidic diet.However the expression of ICAM-1 was infrequent and stable in C57BL/6 mice. Conclusion The expression of VCAM-1 and ICAM-1 in the adventitia of ApoE-/-mice increased gradually with increasing duration of the hyperlipidic diet.

Abstract:Aim To investigate the effect of rosiglitazone(RSG) on glucose-induced adhesion of human endothelial cells.Methods Endothelial cells were incubated with different concentrations of glucose,and then treated with different concentrations of rosiglitazone.Vascular cell adhesion molecule-1(VCAM-1) protein and mRNA expression was detected by immunocytochemical method and RT-PCR.Endothelial-monocyte adhesion assays were carried out in treated cells.Results VCAM-1 mRNA and protein expression were increased significantly with glucose concentration increased,which was obvious at 16.7 mmol/L glucose group(P<0.05).Compared with RSG untreated group,relative VCAM-1 mRNA and protein expression decreased to varying degrees in the RSG treated group(P<0.05).Furthermore,endothelial-monocyte adhesion increased with increment of glucose concentration(P<0.05) and decreased after treatment of high dose RSG(P<0.05).Conclusions RSG may appear to have direct anti-diabetes-associated-atherosclerosis which is at least partly due to effects on VCAM-1 expression induced by glucose,which leads to decreased endothelial-monocyte adhesion and macrophage infiltration.