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.     

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.     

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.     

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.     

Effects of cardiopulmonary bypass on endothelial cell injury

Objective: To observe the changes of plasma concentrations of endotoxin, soluble intercellular adhesion molecule-1, tumor necrosis factor-α, and urinary microalbumin in children undergoing cardiac procedure and to study the effects of cardiopulmonary bypass (CPB) on the injury or activation of endothelial cells and vascular permeability. Methods:Twenty children undergoing cardiac operation with CPB were selected in the study. Plasma concentrations of endotoxin, soluble intercellular adhesion molecule-1, tumor necrosis factor-α, and urinary, microalbumin were measured after anesthetic induction (baseline), bypass for 20 minutes, at the end of CPB, and at 2, 4, and 18 h after the end of CPB. Results: The plasma concentrations of endotoxin, soluble intercellular adhesion molecule-1, and urinary microalbumin began to increase at 2 h after the end of CPB, and remained higher than that of the baseline, while the concentration of tumor necrosis factor-α increased only at the end CPB and at 2 h after the end of CPB. Conclusion: Cardiopulmonary bypass can induce inflammatory response, resulting in the activation or injury of vascular endothelial cells, and can increase the vascular permeability.     

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.     

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     

Polysaccharide sulfate 916 inhibits neutrophil-endothelial adhesion

Objective To study the effect of polysaccharide sulfate 916 (PS916) on neutrophil-endothelial cell adhesion.Methods Cell adhesion was evaluated by testing neutrophil myeloperoxidase activity.Expression of adhesion molecule in human umbilical vein endothelial cell (HUVEC) was measured by ELISA.The neutrophil activation rate induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) was tested by nitroblue tetrazolium (NBT) reduction.Results Tumor necrosis factor alpha (TNFα, 50-800 U/ml) increased the adherence of neutrophil to TNFα-stimulated HUVEC in a concentration and time dependent manner.PS916 (0.01-1.0 mg/ml) dose-dependently inhibited the adherence of neutrophils to TNFα-stimulated HUVEC.fMLP increased the activation rate of neutrophils independent of concentration.PS916 also inhibited the adherence of fMLP-activated neutrophils to HUVEC.Moreover, PS916 inhibited adhesion molecule expression in TNFα-stimulated HUVEC.Conclusions PS916 inhibited neutrophil-endothelial adhesion.The mechanism of its action was partially related to suppressing the expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1)     

Effect of chinese herbal drug-containing serum for activating-blood and dispelling-toxin on ox-LDL-induced inflammatory factors’ expression in endothelial cells

<正>Objective:To investigate the effects of drug-containing serum of Chinese herbal compound, Xiongshao Capsule(芎芍胶囊,XS,for activating-blood) and Huanglian Capsule(黄连胶囊,HL,for dispellingtoxin) on the oxidized low-density lipoprotein(ox-LDL)-induced inflammatory factors in human umbilical vein endothelial cells(HUVECs).Methods:Thirty-two rats were randomly divided into four groups:the blank control group treated with distilled water,the positive control group treated with simvastatin(1.8 mg/kg),the test group I treated with Chinese herbal compound of XS(0.135 g/kg),and the test group II treated with Chinese herbal compound of XS(0.135 g/kg) and HL(0.135 g/kg).All the treatments were administered for 7 successive days by gastrogavage.Rats' blood serum was harvested 1 h after the last administration to prepare respective drugcontaining serum.HUVECs were exposed to ox-LDL(100μg/mL) to induce cell injury model and incubated with corresponding drug-containing serum for 24 h.Untreated HUVECs were set for blank control.Levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and soluble intercellular adhesion molecule-1(slCAM-1) in supernatant of cultured HUVECs were determined by enzyme-linked immunosorbent assay(ELISA).HUVEC surface expressions of ICAM-1 and E-selectin were determined by flow cytometry.Results:Levels of IL-6, TNF-α,and slCAM-1 in the supernatant of HUVECs as well as the cell surface expressions of ICAM-1 and E-selectin significantly increased after 24-h ox-LDL stimulation(P0.01),while the abnormal elevations,except slCAM-1 in the test groupⅠ,were all reduced in the treated groups(the positive control and the two test groups) significantly(P0.01 or P0.05).Besides,the effect in the test group II seemed somewhat higher than that in the test group I but with no statistical significance(P0.05).Conclusion:Drug-containing serum of XS plus HL has a certain inhibitory effect on the vascular endothelial inflammation response induced by ox-LDL.     

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.     

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.     

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     

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.     

Vascular endothelial growth factor up-regulates the expression of intracellular adhesion molecule-1 in retinal endothelial cells via reactive oxygen species, but not nitric oxide

Background The vascular endothelial growth factor （VEGF） is involved in the initiation of retinal vascular leakage and nonperfusion in diabetes. The intracellular adhesion molecule-1 （ICAM-1） is the key mediator of the effect of VEGFs on retinal leukostasis. Although the VEGF is expressed in an early-stage diabetic retina, whether it directly up-regulates ICAM-1 in retinal endothelial cells （ECs） is unknown. In this study, we provided a new mechanism to explain that VEGF does up-regulate the expression of ICAM-1 in retinal ECs. Methods Bovine retinal ECs （BRECs） were isolated and cultured. Immunohistochemical staining was performed to identify BRECs. The cultured cells were divided into corresponding groups. Then, VEGF （100 ng/ml） and other inhibitors were used to treat the cells. Cell lysate and the cultured supernatant were collected, and then, the protein level of ICAM-1 and phosphorylation of the endothelial nitric oxide synthase （eNOS） were detected using Western blotting. Griess reaction was used to detect nitric oxide （NO）. Results Western blotting showed that the VEGF up-regulated the expression of ICAM-1 protein and increased phosphorylation of the eNOS in retinal ECs. Neither the block of NO nor protein kinase C （PKC） altered the expression of ICAM-1 or the phosphorylation of eNOS. The result of the Western blotting also showed that inhibition of phosphatidylinositol 3-kinase （PI3K） or reactive oxygen species （ROS） significantly reduced the expression of ICAM-1. Inhibition of PI3K also reduced phosphorylation of eNOS. Griess reaction showed that VEGF significantly increased during NO production. When eNOS was blocked by L-NAME or PI3K was blocked by LY294002, the basal level of NO production and the increment of NO caused by VEGF could be significantly decreased. Conclusion ROS-NO coupling in the retinal endothelium may be a new mechanism that could help to explain why VEGF induces ICAM-1 expression and the resulting leukostasis in diabetic retinopathy.     

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.     

Levels of soluble adhesion molecules in patients with various clinical presentations of coronary atherosclerosis

Background Adhesion molecules play an important role in the development and progression of coronary atherosclerosis. The aim of this study was to compare concentrations of soluble forms of adhesion molecules in patients with different clinical presentations of coronary artery disease （CAD）.Methods One hundred and twenty-eight patients with CAD were divided into three groups; the first group was acute myocardial infarction group （AMI group, n=45）, the second group was unstable angina pectoris group （UAP group, n=48）,the third group was stable angina pectoris group （SAP group, n=35）. We compared them with patients with normal coronary arteries （control group, n=31）. The serum levels of vascular cell adhesion molecule （VCAM-1）, intercellular adhesion molecule-1 （ICAM-1）, E-selectin and P-selectin were measured in all subjects.Results The serum level of VCAM-1 in the AMI group was significantly higher than in the UAP, SAP and control groups （P 〈0.01）. The level in the UAP group was significantly higher than the SAP group and control group （P 〈0.01） and the level in the SAP group was significantly higher than in the control group （P 〈0.01）. The serum ICAM-1 level was significantly elevated in the AMI, UAP and SAP groups as compared to the control group （P 〈0.01）. The levels of serum E-selectin and P-selectin in the AMI and UAP groups were significantly higher than in the SAP and control groups （P〈0.01）.Conclusions Increased levels of VCAM-1 and ICAM-1, E-selectin and P-selectin, as markers of inflammation, showed the importance of inflammatory processes in the development of atherosclerosis and clinical expression of CAD. Soluble ICAM-1, VCAM-1, E-selectin and P-selectin concentrations are useful indicators of the presence of atherosclerosis and the severity of CAD clinical presentation.     

Mechanism of ligustrazini against thrombosis

Objective To investigate the mechanism of Chinese medicine ligustrazini against thrombosis, and the effects of ligustrazini on plasminogen activator inhibitor (PAI-1) expression in normal endothelial cells and lipopolysaccharide (LPS) stimulated endothelial cells. Methods Human umbilical vein endothelial cells (HUVECs) were cultured by trypsin digestion method. PAI-1 protein in HUVEC conditioned medium was measured by Sandwich enzyme-linked immunosorbent assay (ELISA), and PAI-1 mRNA expression was determined by Northern blot analysis. Using electrophoretic mobility shift assay (EMSA), we observed HUVEC nuclear factor-kappa B (NF-κB) nuclear translocation. Results LPS treatment of cultured HUVECs resulted in a significant increase in PAI-1 protein and mRNA expression by these cells. However, when HUVECs were incubated with LPS plus ligustrazini, the upregulation of PAI-1 by LPS was abated. Moreover, ligustrazini could decrease the basal level of PAI-1 protein and mRNA in HUVECs as compared with control. Nuclear extracts prepared from HUVECs stimulated by LPS demonstrated that binding to the NF-kB oligo nucleotide increased as compared with the unstimulated cells, but ligustrazini did not change those binding in the absence or presence of LPS. Conclusions Ligustrazini inhibited both basal and LPS-induced PAI-1 protein and mRNA expression in endothelial cells, and the modulation of PAI-1 in HUVECs by ligustrazini might have other mechanisms rather than NF-kB     

Inflammatory reaction versus endogenous peroxisome proliferator- activated receptors expression, re-exploring secondary organ complications of spontaneously hypertensive rats

Background The chronic pathological changes in vascular walls of hypertension may exert destructive effects on multiple organ systems. Accumulating evidence indicates that inflammatory reactions are involved in the pathological changes of hypertension. Three peroxisome proliferator-activated receptors （PPARs） have been identified： PPARa, PPARβ/δ, and PPARγ, all of which have multiple biological effects, especially the inhibition of inflammation. The aim of this study was to evaluate PPAR isoforms expression profile in important organs of spontaneously hypertensive rats （SHR） and to understand the modulation of endogenous PPAR isoforms under inflammatory condition. Methods l]ssues （kidney, liver, heart, and brain） were dissected from SHR and age-matched control Wistar-Kyoto rats （WKY） to investigate the abundance of PPAR isoforms and PPAR-responsive genes （acyI-CoA oxidase and CD36）. The expression of CCAAT/enhancer-binding protein 6 （C/EBP6）, which can trans-activate PPARγ expression, was also observed. The inflammatory response was analyzed by the expression of inflammatory mediators inducible nitric oxide synthase （iNOS）, intercellular adhesion molecule-1 （ICAM-1）, vascular cell adhesion molecule-1 （VCAM-1）, E-selectin, interleukin-1 beta （IL-1β）, and tumor necrosis factor alpha （TNFα）, and formation of carbonyl and nitrated proteins. Results The expressions of 3 PPAR isoforms and PPAR-responsive genes were markedly upregulated in SHR compared with those of WKY. Specifically, the expression of PPARa protein in the kidney, liver, heart and brain increased by 130.76 %, 91.48%, 306.24%, and 90.70%; PPARβ/δ upregulated by 109.34%, 161.98%, 137.04%, and 131.66%; PPARγ increased by 393.76%, 193.17%, 559.29%, and 591.18%. In consistent with the changes in PPARy, the expression of C/EBPδ was also dramatically elevated in SHR. Inflammatory mediators expressions were significantly increased in the most organs of SHR than WKY. As a consequence, increased formation of