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 The effect of oxidized low density lipoprotein （ox-LDL） on proaggregation and proadhesion in human umbilical vein endothelial cells （HUVEC） and the underlying mechanisms were observed. Methods HUVEC in 6-well plate were treated with varied concentrations of ox-LDL （ 50, 100 and 200 mg/L） for 12 h, 24 h and 48 h, respectively. After that, the levels of prostacyclin （PGI2） and thromboxane A2 （TXA2） in supernatants were detected by radioimmunoasssay. Finally, the expression levels of tissue factor （TF）, intercellular cell adhesion molecule-1 （ICAM-1） and vascular cell adhesion molecule-1 （VCAM-1） were examined by Western blot. Results Dose-response assay showed that the proaggregation of HUVEC induced by ox-LDL was enhanced in a concentration-dependent manner by treatment with ox-LDL for 24 h, presenting that the ratios of PGI2/TXA2 markedly decreased （P<0.05 or P<0.01）. The similar results from time-course relationship assay were obtained. In addition, the expression levels of TF significantly increased in a fashion of concentration-dependence. More importantly, the proadhesion of HUVEC was enhanced, displaying the expression levels of ICAM-1 and VCAM-1 were significantly increased in a concentration-dependent fashion. Conclusions ox-LDL enhances the activities of proaggregation and proadhesion in HUVEC in a concentration-dependent manner. The underlying mechanisms might downregulate the value of PGI2/TXA2 and upregulate the expression of TF, ICAM-1 and VCAM-1.

Abstract:AimTo compare alliin and allicin’s effect in inhibiting tumor necrosis factor-α(TNF-α) induced inflammation, and to investigate whether alliin can substitute allicin in resisting atherosclerosis.MethodsHuman umbilical vein endothelial cell (HUVEC) were divided into ten groups and treated for 16 hours as follows:Control(treated without any drugs), TNF-α(1 μg/L), alliin (10 mg/L), allicin (10 mg/L), TNF-α(1 μg/L) and alliin (5 mg/L,10 mg/L,30 mg/L),TNF-α (1 μg/L) and allicin(5 mg/L,10 mg/L,30 mg/L).The mRNA expression level of intercellular cell adhesion molecule-1 (ICAM-1) was determined by real-time PCR.THP-1 adhesion to HUVEC was measured by rose Bengal staining.Migration function and MTT text were used to demonstrate the effects of alliin and allicin on the smooth muscle cell’s migration and proliferation.ResultsTNF-α treatment significantly increased the mRNA expression of ICAM-1 in HUVEC, while alliin and allicin pretreatment all blocked the TNF-α induced mRNA expression of ICAM-1, and also effectively blocked THP-1 adhesions.Further more, alliin and allicin inhibited TNF-α induced contractile fiber cell’s migration and proliferation functions.However, allicin was a bit more effective than alliin.ConclusionBoth alliin and allicin can inhibit TNF-α induced inflammation and atherosclerosis, and alliin may substitute allicin in resisting atherosclerosis to a certain extent.

Abstract:AimTo investigate the molecular mechanism of inflammatory protein expression via transforming growth factor-β1 (TGF-β1)/Smad pathway in ox-LDL-stimulated human umbilical vein endothelial cells (HUVEC).MethodsHUVEC were treated with 0.1 g/L of ox-LDL.TGF-β1, Smad3, VCAM-1 and ICAM-1 protein expression, and Smad3 phosphorylation were detemined by Western blot.Resultsox-LDL obviously increased the expression of VCAM-1 and ICAM-1 in HUVEC.TGF-β1 expression and Smad3 phosphorylation were enhanced in ox-LDL-stimulated HUVEC.Nuclear extract analysis showed that the phosphorylated Smad3 expression was increased in ox-LDL-treated cells.SIS3 (a specific inhibitor of Smad3) inhibited Smad3 phosphorylation in a dose-dependent manner, however, VCAM-1 and ICAM-1 expression were increased in ox-LDL-treated cells.ConclusionsTGF-β1/Smad signaling involved in the regulation of inflammatory protein expression in HUVEC induced by ox-LDL, the inhibition of Smad3 phosphorylation increased the expression of inflammatory protein, suggesting that TGF-β1/Smad3 signaling repressed the inflammatory protein expression in ox-LDL-treated endothelial cells.

Abstract:Aim To explore the time course of expression of ICAM-1 gene and protein in macrophages foamy process induced by ox-LDL. Methods Cell cycle of Human myeloid leukemia U937 cells was arrested in G1 by preincubated with thymidine (2.0 mmol/L) for 24 h, then cells released at the same time. U937 cells were incubated with ox-LDL (80 mg/L) for time course(12 h, 24 h, 48 h or 72 h) treatment. Results Flow cytometric analysis results revealed that ICAM-1 protein expression increased from 12 h to 24 h and was highest at 24 h, but ICAM-1 decreased at 48 h. RT-PCR results showed that ICAM-1 mRNA level increased consistently with protein expression. Conclusions These results proved that ox-LDL could increase ICAM-1 gene and protein expression on U937 cells. Accumulation of cholesteryl esters in U937 cells has no negative feedback control on ICAM-1 gene and protein expression.