Ginsenoside Rg1-induced alterations in gene expression in TNF-α stimulated endothelial cells

Background In China the ginseng root began to be used in medicine over 2000 years ago. Ginsenosides are the most important component isolated from ginseng. The authors investigated the effect of ginsenoside Rg1 on the spectrum of gene expression in the endothelial cells stimulated by TNF-α and further explored the potential molecular mechanism of endothelial protection by ginsenoside Rg1.
Methods Nitric oxide (NO) production in the cultured human umbilical vein endothelial cells(HUVECs) was measured by using an NO assay kit. A home-made oligonucleotide microarray containing approximately 400 cardiovascular disease-related genes was constructed. The alteration of the spectrum of gene expression induced by ginsenoside Rg1 in HUVECs which were activated by TNF-α were detected by oligonucleotide microarray analysis.
Results NO production in HUVECs was decreased significantly after TNF-α treatment, while pretreatment with ginsenoside Rg1 enhanced NO production in TNF-αstimulated HUVECs. Ginsenoside Rg1 affected the expression levels of genes involved in vascular constriction, cell adherence, coagulation, cell growth and signal transduction in TNF-αstimulated HUVECs.
Conclusions Ginsenoside Rg1 could enhance NO production and the expression of eNOS mRNA in TNF-α stimulated HUVECs. Ginsenoside Rg1 regulated sets of genes in endothelial cells and protected endothelial cells from TNF-αactivation. Microarray analysis provided us with valuable insights into the atheroprotective mechanism by gingsenoside Rg1.
     

Pingyangmycin-regulated expressions of adhesion molecules in human venous malformation endothelial cells

Pingyangmycin (bleomycin A5 hydrochloride,PYM) is one of the anti-neoplastic agents which have been commonly used to treat venous malformations.However,the underlying mechanism by which PYM treats venous malformations remains poorly understood.It was reported that venous endothelial cells could recruit neutrophils via adhesion molecules (E-selectin,ICAM-1,ICAM-3,VCAM-1) during the acute/chronic inflammation and subsequent histological fibrosis after sclerotherapy with PYM.This study explored if the expression of E-selectin,ICAM-1,ICAM-3 and VCAM-1 in human venous malformation endothelial cells could be affected by PYM.HVMECs were cultured from human venous malformation tissue.Expressions of E-selectin,ICAM-1,ICAM-3 and VCAM-1 on HVMECs in response to PYM were analyzed by cell ELISA.The relative levels of mRNA expression in the cells were semi-quantified.The results showed that PYM up-regulated the expressions of E-selectin,ICAM-3,VCAM-1 and ICAM-1 in both time-and concentration-dependent manner.Our findings suggested that PYM could induce the expression of adhesion molecules in HVMECs,which might be a possible mechanism by which sclerotherapy by intralesional injection of PYM treats venous malformations.     

Angiogenic Effect of Intercellular Adhesion Molecule-1

In order to investigate the angiogenic effect of intercellular adhesion molecule-1 (ICAM-1), two parts of experiment were performed. Chick embryo chorioallantoic membrane (CAM) assay was used for in vivo angiogenic research. The chick embryos were divided into 4 groups: ICAM-1 group (divided into 3 subgroups, Ⅰ, Ⅱ and Ⅲ) for screening the angiogenic effect of ICAM-1 by adding different concentrations of ICAM-1 (0.1, 0.2 and 0.3 μg/μL) 5 μL into the chick embryo CAMs on the day 10 after incubation for every subgroup; Anti-ICAM-1 group A (divided into 2 subgroups, Ⅰ and Ⅱ) by adding different concentrations of Anti-ICAM-1 (1:100, 1:50) 5μL into the chick embryo CAMs on the day 10 after incubation for every subgroup to evaluate the effect of ICAM-1 on the survival of microvessels through observing whether Anti-ICAM-1 could induce involution of the microvessels on CAMs; Anti-ICAM-1 group B (divided into 2 subgroups, Ⅰ and Ⅱ) by adding different concentrations of Anti-ICAM-1 (1:100, 1:50) 5 μL into the chick embryo CAMs on the day 6 after incubation for every subgroup to evaluate whether ICAM-1 involved in embryonic angiogenesis through observing the growth of microvessels on CAMs; Control group: ICAM-1 or Anti-ICAM-1 was substituted by PBS 5 μL on the day 10 or day 6 after incubation. Three days later, the CAMs were photographed in vivo, excised, sectioned and the number of microvessels was counted. In ICAM-1 group, there was increased number of microvessels arranged radially with "spoked-wheel" pattern around the gelatin sponges. The new microvessels growing perpendicularly to gelatin sponges were observed. The number of the microvessels growing in the CAM mesen-chymes around the sponges in 3 subgroups was higher than that in control group (P<0.01), however, there was no significant difference among the 3 subgroups (P>0.05). In anti-ICAM-1 group A, the radially arranged microvessels were very unclear around the sponges contrast to that of ICAM-1 group. Few new microvessels were detected in the center of the sponges. The number of the microvessels growing in the CAM mesenchymes around the sponges in subgroup Ⅱ was lower than that in control group (P<0.01). There was no significant difference in the number of the microvessels around the sponges between subgroup Ⅰ and control group (P>0.05). In anti-ICAM-1 group B, the radially arranged microvessels were very unclear around the sponges contrast to that of control group. New microvessels were very scarce in the center of the sponges.The number of the microvessels growing in the CAM mesenchymes around the sponges in the 2 subgroups were less than that in control group (P<0.01), and there was significant difference between the 2 subgroups (P<0.05). It was suggested that ICAM-1 could induce angiogenesis and support the survival of microvessels, and ICAM-1 was involved in embryonic angiogenesis.     

Tanshinone ⅡA Protects against Lipopolysaccharides-lnduced Endothelial Cell Injury via Rho/Rho Kinase Pathway

Objective:To test whether tanshinone ⅡA(Tan ⅡA),a highly valued herb derivative to treat vascular diseases in Chinese medicine,could protect endothelial cells from bacterial endotoxin(lipopolysaccharides,LPS)-induced endothelial injury.Methods:Endothelial cell injury was induced by treating human umbilical vein endothelial cells(HUVECs) with 0.2 μg/mL LPS for 24 h.Y27632 and valsartan were used as positive controls.The effects of tanshinone TJ A on the LPS-induced cell viability and apoptosis rate of HUVECs were tested by flow cytometry,cell migration by transwell,adhesion by a 96-well plate pre-coated with vitronectin and cytoskeleton reorganization by immunofluorescence assay.Rho/Rho kinase(ROCK) pathwayassociated gene and protein expression were examined by microarray assay;quantitative real-time polymerase chain reaction and Western blotting were used to confirm the changes observed by microarray.Results:Tan ⅡA improved cell viability,suppressed apoptosis and protected cells from LPS-induced reductions in cell migration and adhesion at a comparable magnitude to that of Y27632 and valsartan.Tan ⅡA,Y27632 and valsartan also normalized LPS-induced actomyosin contraction and vinculin protein aggregation.A microarray assay revealed increased levels of fibronectin,integrin A5(ITG A5),Ras homolog gene family member A(RhoA),myosin light chain phosphatase,phosphatidylinositol-4,5-bisphosphate 3-kinase(PI3K,or PIP2 in Western blotting),focal adhesion kinase,vascular endothelial growth factor and vascular endothelial growth factor receptor 2 in the damaged HUVECs,which were attenuated to different degrees by Tan ⅡA,Y27632 and valsartan.Conclusion:Tan ⅡA exerted a strong protective effect on HUVECs,and the mechanism was caused,at least in part,by a blockade in the Rho/ROCK pathway,presumably through the down-regulation of ITG A5.     

Expression of Intercellular Adhesion Molecule-1 in Immune Response of Inner Ear

To understand the role of intercellular adhesion molecule-1 (ICAM-1) in immune response of the inner ear, inner ear immune response was induced in rats by inoculation of keyhole limpet hemocyanine (KLH) into the scala tympani of the animals who had been systemically sensitized. The expression of ICAM-1 in the inner ear was immunohistochemically examined. ICAM-1 was found in the epithelium of the spiral modiolar vein (SMV) with its collecting venules (CVs) as early as 6 h after challenge. Expression of ICAM-1 was observed on the epithelium of the endolymphatic sac (ES) and perisaccular region at 12 h. The intensity of ICAM-1 staining reached its peak within 24-48 h in these sites of the inner ear. By day 28, most specimens were devoid of appreciable staining for ICAM-1.Our study demonstrates that adhesion molecules play an important role in extravasation of inflammatory cells from the systemic circulation in the process of inner ear immune response. It also shows that cytokines that control expression of     

Effects of static magnetic field on human umbilical vessel endothelial cell

Objective： To investigate the effects of static magnetic field（SMF） on the viability, adhesion molecule expression of human umbilical vessel endothelial cell. Methods： Magnetic flux intensity was 0. 1 mT, 1 mT, 10 mT. Cell viability and proliferation were measured with ^3H-TdR and MTT methods; and apoptosis of human umbilical vein endothelial cell （HUVEC） was studied by flow cytometry and transmission electric microscopy. ELISA was used to measure the expression of ICAM-1 and VCAM-1 on endothelium. Results： 0.1 mT SMF had no effects on the growth of HUVEC, however, SMF of 1 mT, 10 mT attenuated growth of HUVEC. 10 mT static magnetic field could induce apoptosis and necrosis of HUVEC. 10 mT SMF enhanced the expression of ICAM-1 and VCAM-1 on endothelium. Conclusion： The effect of SMF depends on the intensity of SMF. 10 mT SMF has adverse effects on human umbilical vessel endothelial cell.     

Soluble cell adhesion molecules in patients with acute coronary syndrome

Objective To observe the changes in serum soluble intercellular adhesion molecule type-1(ICAM-1), vascular cell adhesion molecule type-1(VCAM-1), E-selectin and von Willebrand factor (vWf) in patients with acute coronary syndrome. Methods Serial venous blood samples were taken from 21 patients with acute myocardial infarction(AMI) before and 4, 8, 12, 24, 48 and 72 h after thrombolytic treatment or direct PTCA. One blood sample was drawn from 16 patients with unstable angina and 16 control subjects. Serum concentrations of ICAM-1, VCAM-1, E-selectin and vWf were determined using a double antibody sandwich enzyme-linked immunosorbent assay. Results Serum levels of ICAM-1, VCAM-1, E-selectin and vWf were higher in patients with acute coronary syndrome than in controls. Patients with AMI and successful reperfusion therapy had a significant reduction in the serum concentration of ICAM-1 and E-selectin at 24 and 48 h, VCAM-1 at 24 and 72 h and vWf at 12,　24,　48 and 72 h, but had peak in serum levels of ICAM-1 and E-selectin at 4 h. The number of diseased coronary arteries was not related to the levels of ICAM-1,　VCAM-1 and E-selectin. Conclusion The serum concentration of soluble cell adhesion molecules was elevated significantly in patients with acute coronary syndrome. Successful reperfusion therapy was associated with a reduction in the serum concentrations of soluble cell adhesion molecules in patients with AMI.     

Influence of traditional Chinese medicine Qingjieling () on LFA-1-dependent leukocyte adhesion to endothelial cells) on LFA-1-dependent leukocyte adhesion to endothelial cells

Objective: To investigate the influence of traditional Chinese medicine Qingjieling on leukocyte adhesion to vascular endothelial cells and the possible mechanisms.Methods: Firstly, We successfully established in vitro culture of human umbilical vein endothelial cells (HUVEC) monolayer. Then we constructed a leukocyte-endothelial cell adhesion model and imitated the inflammation state by using endothelial cells stimulated with recombinant tumor necrosis factor a (TNFα) or interleukin-1 (IL-1) respectively. The drug-induced change of leukocyte adhesion function was observed after we directly administered traditional Chinese medicine Qingjieling on the leukocytes. Finally, we inquired into the molecular mechanism of the effect of Qingjieling by monoclonal antibody blocking assay.Results: HUVEC as a representative large vessel endothelial cell could perfectly construct an adhesion model with leukocytes. Under the stimulation of cytokines (TNFα or IL-1), endothelial cells demonstrated drastically improved adhesion with leukocytes, certain dose of Qingjieling (1mg/g body weight) might augment the leukocyte adhesion capacity with a 2-hour co-incubation. At least part of the reason of Qingjieling’s improving adhesion effect is due to the increased lymphocyte functional associated antigen-1 (LFA-1) expression of the white blood cells.Conclusion: Qingjieling could increase LFA-1-dependent leukocytes adhesion to endothelial cells.     

Cardiotrophin-1 induces intercellular adhesion molecule-1 expression by nuclear factor κB activation in human umbilical vein endothelial cells

Background In addition to elevated concentrations of cytokines, patients with congestive endothelial dysfunction and increased plasma concentrations of adhesion molecules heart failure （CHF） show ke intercellular adhesion molecule-1 （ICAM-1）. Furthermore, the concentration of cardiotrophin-1 （CT-1） - a cytokine of the interleukin-6 superfamily - is increased in CHF. We tested the hypothesis whether CT-1 is able to induce ICAM-1 in human umbilical vein endothelial cells （HUVEC）. Furthermore we examined the signalling mechanisms of CT-1 mediated ICAM-1 expression. Methods Confluent layers of HUVEC were incubated with increasing concentrations of CT-1 （5 to 100 ng/ml） for different periods. ICAM-1 mRNA was determined by real-time polymerase chain reaction （PCR） and ICAM-1 surface expression by fluorescence-activated cell sorter （FACS） analysis and soluble ICAM-1 （slCAM-1） in the culture supernatant by enzyme linked immunosorbent assay （ELISA）. To clarify the signalling pathway of CT-1 induced ICAM-1 expression we used various inhibitors of possible signal transducing molecules, electromobility shift assay （EMSA） and Western blot analysis. Results CT-1 induced ICAM-1 mRNA （1.8±0.8 fold increase compared to unstimulated cells after 6 hours） and protein （1.4±0.2 fold increase compared to unstimulated cells after 48 hours） in HUVEC in a time- and concentration-dependent manner. EMSA experiments show that CT-1 causes nuclear factor （NF） κB activation. Because parthenolide could inhibit CT-1 induced ICAM-1 expression NFκB activation is required in this pathway. CT-1 did not activate extracellular signal regulated kinases （ERK）, c-Jun N-terminal kinase （JNK） and p38. Conclusion CT-1 is able to induce ICAM-1 in endothelial cells by NFκB activation. These results may explain in part elevated ICAM-1 concentrations in patients with CHF and endothelial dysfunction.     

Correlation of integ rin α4β1 and its two ligands with mast cell recruitment around the rat liver neoplasm

Aim To study the correlation of integfin α4β1and its twoligands (vascular cell adhesion molecule-1 and fibronectin)with mast cell (MC) recruitment around the rat liverneoplasm.Methods 18 male wistar rats with liver tumor were dividedinto three different groups in terms of mast cell number inthe su .rroundings of liver tumor, 8 normal wistar rats ascontrol. The integrin VLA-4 expression of rat peritoneal mastcells was analyzed by indirect immunofluorescence and flowcytometry. We also used immunohistochemistry to investigatewhether VCAM-1 and fibronectin in liver tissues wereexpressedpositively.Results There were markedly different in mast cell numberaround rat liver neoplasms. And mast cells could expresshigh levels of integrin α4β1 on their surfaces. Furthermor,the more mast cells around liver tumor the higher levels ofintegrin VLA-4. We also found that endothelial cellsexpressed VCAM-1 and there are a number of fibronectindeposition aroundrat fiver neoplasm.Conclusion The results suggest that the integrin α4β1/VCAM-1 and fibronectin play an important role inmechanism of mast cell recruitment around liver tumor. Andthe expression levels of integrin α4β1 were paralleled by mastcell accumulation in the surroundings of liver neoplasm.     

Protective effects of Radix Astragalus on vascular endothelial Cells of patients with Binswanger’s disease

Objective: To compare the effects of Radix Astragalus (RA) on vascular endothelial cells in Binswanger’s disease (BD) patients with Radix Salviae miltiorrhizae (RSM).Methods: There were 37 patients with BD in the treated group and 37 healthy subjects in the control group. Thirty-seven patients were further randomly subdivided into two groups: RA group (19 patients) and RSM group (18 patients). Circulating endothelial cells (CEC) and the levels of endothelin-1 (ET-1), nitric oxide (NO) and malondialdehyde (MDA) in the blood of internal jugular vein which were examined before and after treatments.Results: When compared with those of the control group, CEC counts, ET-1 and MDA levels in plasma increased significantly, meanwhile serum NO concentration decreased significantly in the treated group. When compared with those of pretreatment, CEC counts, ET-1 and MDA decreased significantly and serum NO concentration increased significantly after treatment in RA group. There were no significant changes of these indices in RSM group after treatment.Conclusions: There are damage and dysfunction of vascular endothelial cells in patients with BD. RA injection is an effective drug to protect vascular endothelial cells of BD patients.     

Cationic antimicrobial protein of Mr 37 kDa: a multifunctional inflammatory protein

Purpose　To investigate the role of cationic antimicrobial protein of Mr 37?kDa (CAP37) a neutrophil-derived inflammatory mediator on endothelial cell function. Data sources　Endothelial cells used in this study were obtained from human lung microvessels and rat aorta. The latter was a kind gift of Dr. Paula Grammas. The mono-mac 6 cell line used in this study was the generous gift of Dr. H.W. Loms Ziegler-Heitbrock. Study selection and data extraction　Endothelial cell proteins kinase C activity was determined by measuring calcium- and phospholipid-dependent phosphorylation of histone. Endothelial cell migration was determined using CostarTM Transwell apparatus. Cell surface expression of adhesion molecules, ICAM-1 and PECAM-1 was determined using flow cytometry. RT-PCR was used to amplify the CAP37 from endothelial cells treated with LPS. Results　We demonstrated that CAP37 which was originally identified as having potent antimicrobial activity and chemotactic activity for monocytes was capable of modulating endothelial cell functions. CAP37 activated endothelial cell protein kinase C in a dose- and time-dependent fashion. Importantly CAP37 increased the adhesive properties of the endothelium for monocytes. CAP37 upregulated the well known adhesion molecules, ICAM-1 and PECAM-1 in a dose- and time- dependent manner. In addition, CAP37 promoted endothelial cell migration. Further investigations indicated that CAP37 was induced in endothelial cells in response to pro-inflammatory cytokines such as tumor necrosis factor-α and interleukin-1α as well as inflammatory mediators such as lipopolysaccharide. Unstimulated endothelial cells did not constitutively express CAP37. The cDNA sequence of endothelial CAP37 was determined and found to be highly homologous to the sequence obtained for neutrophil-derived CAP37. Conclusions　Our studies strongly suggest that CAP37 plays a pivotal role in monocyte-endothelial interactions and the transmigration of monocytes from the vasculature into extravascular tissues.     

Comparison of the effects of recombinant human endostatin and docetaxel on human umbilical vein endothelial cells in different growth states

Background  Recombinant human endostatin (rh-endostatin, Endostar) has been proved to be an inhibitor of angiogenesis. Docetaxel has been also considered as a common chemotherapeutic agent with inhibition of angiogenesis of malignancies. However, their function has been seldom compared and a best synergism protocol is not determined. This study aimed to compare the effects of two drugs, investigate their combined impact on human umbilical vein endothelial cells (HUVECs), a molecular basis and find ideal protocols to inhibit endothelial cell proliferation.

Methods  HUVECs on confluent growth or activated by vascular endothelial growth factor (VEGF) were treated by rh-endostatin or/and docetaxel at respective gradient concentration in following operations as cell proliferation determined by MTT assay, cell cycle distribution, apoptosis and markers of CD146, CD62E and CD105 detected by flow cytometery, the structure of the channel formed by HUVECs measured by tube formation count.

Results  Rh-endostatin exhibited time dependent inhibition of proliferation while docetaxel showed both time and dose dependent inhibition. HUVECs accumulated in G₀-G₁ with decreased numbers of cells in G₂ after a single treatment of rh-endostatin or that followed by docetaxel treatment. Cells accumulated in G₂ after both a single docetaxel and simultaneous administration. Both the number of cells in G₀-G₁ and apoptotic cells were increased by docetaxel followed by rh-endostatin treatment. The number of non-apoptotic cells at G₀-G₁ was increased by first administering rh-endostatin then docetaxel. Sequential treatment of docetaxel followed by rh-endostatin resulted in the greatest increase in apoptosis (34.7%) and the second highest apoptosis was seen with simultaneous administration (18.2%). Expression of CD146 and CD105 on confluent HUVECs was reduced at certain doses of rh-endostatin and/or docetaxel. However, rh-endostatin reduced CD105 without any apparent impact on either CD146 or CD62E expression, whereas these markers were down-regulated by docetaxel after pre-activation by VEGF. Rh-endostatin treatment maintained tube-like structures for a limited time. In contrast, docetaxel swiftly reduced tube formation. Simultaneous treatment, or docetaxel followed by rh-endostatin, exhibited a stronger inhibition on tube formation than either agent alone.

Conclusions  Both rh-endostatin and docetaxel can inhibit HUVEC proliferation while the high apoptotic rate after combined administration was probably owing to different sequent administration by docetaxel followed by rh-endostatin or simultaneous treatment. Both proliferation and adhesion molecules on HUVECs of confluent growth are down-regulated by the two drugs. The rh-endostatin decreased proliferation markers, but only slightly modified adhesion molecules, while both markers were down-regulated by docetaxel on HUVECs activated by VEGF. Rh-endostatin could maintain adhesion of HUVECs at first then induce cells apoptosis to damage tube formation. We hypothesize that it could lead to vascular normalization in short time. In contrast, docetaxel can suppress HUVEC proliferation, adhesion, and reduced tube formation swiftly due to its cytotoxicity. Combined treatments can induce a synergistic inhibition of tube formation.

     

Rat Polymorphonuclear Leukocyte-Endothelium Adhesion Decreases the Permeability of Endothelial Monolayers

The effects of polymorphonuclear leukocyte(PMN)-endothelial cell(EC) adhesion on the permeability of microvascular endothelial monolayers to fluid and albumin were investgated. It was found that: 1)the adherence of fresh PMNs to EC reduced the permeability of untreated endothelial moralayers; 2)the dherence of fresh PMN to platelet activating factor(PAF)-activated endothelial monolayers reduce(?) AF-induced high permeability     

丹参酮ⅡA通过 Rho/Rho激酶途径保护脂多糖诱导内皮细胞损伤

Objective： To test whether tanshinone ⅡA （Tan ⅡA）, a highly valued herb derivative to treat vascular diseases in Chinese medicine, could protect endothelial cells from bacterial endotoxin （lipopolysaccharides, LPS）-induced endothelial injury. Methods： Endothelial cell injury was induced by treating human umbilical vein endothelial cells （HUVECs） with 0.2 μg/mL LPS for 24 h. Y27632 and valsartan were used as positive controls. The effects of tanshinone Ⅱ A on the LPS-induced cell viability and apoptosis rate of HUVECs were tested by flow cytometry, cell migration by transwell, adhesion by a 96-well plate pre-coated with vitronectin and cytoskeleton reorganization by immunofluorescence assay. Rho/Rho kinase （ROCK） pathway- associated gene and protein expression were examined by microarray assay; quantitative real-time polymerase chain reaction and Western blotting were used to confirm the changes observed by microarray. Results： Tan ][ A improved cell viability, suppressed apoptosis and protected cells from LPS-induced reductions in cell migration and adhesion at a comparable magnitude to that of Y27632 and valsartan. Tan II A, Y27632 and valsartan also normalized LPS-induced actomyosin contraction and vinculin protein aggregation. A microarray assay revealed increased levels of fibronectin, integrin A5 （ITG A5）, Ras homolog gene family member A （RheA）, myosin light chain phosphatase, phosphatidylinositol-4,5-bisphosphate 3-kinase （PI3K, or PIP2 in Western blotting）, focal adhesion kinase, vascular endothelial growth factor and vascular endothelial growth factor receptor 2 in the damaged HUVECs, which were attenuated to different degrees by Tan ⅡA, Y27632 and valsartan. Conclusion： Tan ⅡA exerted a strong protective effect on HUVECs, and the mechanism was caused, at least in part, by a blockade in the Rho/ROCK pathway, presumably through the down-regulation of ITG A5.     

Effect of Troglitazone on Expression of Adhesion Molecules and eNOS in Human Saphenous Vein Gaft

To investigate whether peroxisome proliferators-activated receptor-γ (PPARγ) ligand Troglitazone can reduce endothelial injury and activation during storage of harvested saphenous vein grafts. Segments of human saphenous vein graft were collected from 9 patients undergoing coronary bypass surgery and then divided into two equal parts of control and test specimens, were stored in ei- ther heparinized blood (control group) or heparinized blood containing 20 μmol/L troglitazone (test group) for 1 h at room temperature. Tissue distribution and protein expression of VCAM-1, ICAM-1, and endothelial nitric oxide synthase (eNOS) were compared using immunohistochemistry and West- ern blot analysis. Myeloperoxidase (MPO) activity, a marker of neutrophil sequestration in human saphenous vein grafts, was also measured in each group. The expression of ICAM-1 (753±132 versus 7201±934; P<0.01),VCAM-1 (3731±294 versus 8292±793; P<0.01), and MPO activity (1.52±0.42 U/g,5.04±1.26 U/g P<0.01) were significantly lower in test group. In contract, eNOS expression (7983±834 versus 3989±1008; P<0.01) was significantly higher in test group. PPARγ ligand troglita- zone might reduce endothelial injury during the storage period of human saphenous vein grafts.     

Influence of glycated low density lipoprotein on the proliferation, expression of intercellular adhesion molecule-1, von Willebrand factor of human umbilical endothelial cells

Diabetes mellitus known as its macro- and microangiopathy has caused thousands of mortality per year. Recent researches showed that hyperglycemia, advanced glycation end products （AGEs） and some other factors acted on the process of atherogenesis. AGEs can combine with receptors of AGEs （RAGEs）, which exist on the vascular endothelium, smooth muscle cells, macrophage, lymphocyte and so on. They can stimulate series of signal transduction systems including nuclear factor κB （NF-κB） pathway, finally promote the secretion of inflammatory factor such as interleukin-1 （IL-1）, interleukin-6 （IL-6）, tumor necrosis factor-α （TNF-α）, intercellular adhesion molecule-1 （ICAM-1）,1＇2 as well as increase the synthesis and secretion of coagulant modulatory factors such as vascular cell adhesion molecule- 1 （VCAM- 1） and von Willebrand factor （vWF）.3     

Anisodamine Inhibits Engotoxin—induced Tissue Factor Expression in Human Endothelial Cells

By study on the effect of anisodamine on lipopolysaccharide-induced expression of tissue factor(TF) in vascular endothelial cells (EC),the mechanism of anisodamine antithrombosis,as well as in the treatment of bacteraemic shock was investigated.Human umbilical vein endothelial cells (HUVECs) were cultured by trypsin digestion method.TF activity was measured in the lysates of HUVEC by using a single step clotting assay.Specific mRNA expression was detected by Northern blotting.In order to evaluate a possible contribution of the nuclear factor (NF)-κB pathway on the effects observed,electrophoretic mobility shift assays (EMSA) were performed using nuclear extracts from HUVECs and NF-κB-binding oligonucleotides.The results showed that treatment of HUVEC with LPS resulted in a significant increase in TF activity.Anisodamine dose-dependently inhibited LPS-induced upregulation of TF.These effects was also confirmed on the level of specific TF mRNA expression by Northern blotting.Furthermore,EMSA showed that anisodamine completely abolished LPS-induced NF-κB DNA binding activity in nuclear extracts from HUVECs treated with LPS together with anisodamine.The results suggest that anisodamine counteracts endothelial cell activation by inhibiting LPS-induced TF expression in these cells.Its interference with the NF-κB pathway might-at least in part-contribute to this effect.The ability of anisodamine to counteract LPS effect on endothelial cells might be one underlying mechanism explaining its antithrombosis and efficacy in the treatment of bacteraemic shock.