Advanced glycation end products-modified protein up-regulates expression of adhesion molecules in human umbilical vein endothelial cells

AIM: Advanced glycation end products (AGEs)-modified proteins are found in plasma and atherosclerosis lesion of diabetes mellitus patients. The study was conducted to elucidate the effect of AGE on expression of adhesion molecules in human endothelial cells. METHODS: Human endothelial cells derived from umbilical veins (HUVECs) were coincubated in vitro with native human serum albumin (HSA) or HSA modified with advanced glycation end products (AGE-HSA). The expression of adhesion molecule intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were determined by immunofluorescent staining and flow cytometry analysis. RESULTS: ICAM-1 and VCAM-1 were constitutively expressed on HUVECs. AGE-HAS enhanced the expression of ICAM-1 and VCAM-1 on HUVECs in a time- and dose-dependent manner. HSA had no effect on the expression of adhesion molecules. CONCLUSIONS: AGE-HSA up-regulates the expression of adhesion molecules in human endothelial cells. AGEs may therefore promote the infiltration of monocytes in the vascular endothelium and have an important effect in the generation and progress of atherosclerosis.     

Paeonol reduces expression of adhesion molecules in HUVECs induced by hyperlipidemic serum via inhibiting the pathway of NF-κB signaling

AIM: To observe the anti-atherosclerosis effect of paeonal (Pae) on the activation of NF-κB and the expression of cell adhesion molecules in human umbilical vein endothelial cells (HUVECs) induced by hyperlipidemic serum. METHODS: Cultured HUVECs were used as target cells. Hyperlipidemic serum was added to the culture medium to establish the injury mode of HUVECs. Methyl thiazolyl tetrazolium (MTT) method was used to examine the cell viability. The mRNA expression of NF-κB p65 was determined by RT-PCR. The protein levels of IκB-α, intercellular cell adhesion molecule-1 (ICAM-1) and E-selectin were detected by Western blotting. RESULTS: After treated with Pae, the cell viability was increased and the morphological changes of HUVECs injured by hyperlipidemic serum trended to normal. The expression of IκB-α in HUVECs injured by hyperlipidemic serum increased, while the expression of NF-κB p65 mRNA, ICAM-1 and E-selectin protein was decreased. CONCLUSION: The anti-atherosclerosis mechanism of paeonal may be related to the inhibitory effect of the natural compound on the pathway of NF-κB/IκB, thereby reducing the expression of ICAM-1 and E-selectin and attenuating the inflammatory reaction in vascellum.     

Effect of mininally modified low density lipoprotein on the adhesive function of vascular endothelial cell and its mechanism

AIM: To observe the effect of MM-LDL (minimally modified LDL) on the interaction between human umbilical vein endothelial cell (HUVEC) and U937 monocyte-like cell line and the exporession of vascular cell adhesion-1 (VCAM-1), intercellular adhesion molecule-1(ICAM-1), P-selectin.METHODS:The adhesive percentage between HUVEC treated with MM-LDL and U937 was determined by counting and the expression of VCAM-1, ICAM-1, P-selectin were examined by ELISA. RESULTS: Treatment of HUVEC with MM-LDL (75 mg/L) for 4 hours significantly increased adhesion of U937 to HUVEC ( P <0.01) and did not induce the surface expression of VCAM-1, ICAM-1, P-selectin . Recombination tumor necrosis factor-alpha (rTNF_α) 5.0 μg/L, as a positive control, induced the expression of these adhesion molecules ( P <0.05). Prolonged (18 h) exposure to MM-LDL resulted in the expression of P-selectin, but not VCAM-1.CONCLUSION: the adhesion of monocytes to endothelial cells induced by MM-LDL is not mediated by VCAM-1, ICAM-1. P-selectin induction may be partly involved in the process.     

Effect of water extract propolis on expression of vascular endothelial cell adhesion molecules

AIM: To explore the mechanism of propolis on the inhibition of atherosclerosis and thrombosis in injured human umbilical vascular endothelial cells (HUVECs) induced by tumor necrosis factor alpha (TNF-α)in vitro.METHODS: TNF-α at the concentration of 50 μg/L was used to induce the injury of HUVECs. The injured HUVECs were treated with water extract propolis (WEP) at the concentrations of 50, 100 and 200 mg/L for 6 h, 12 h and 24 h. The expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) was examined by flow cytometry.RESULTS: The expression of ICAM-1 and VCAM-1 was significantly higher in injured HUVECs (P<0.01) than that in the control cells. The expression of ICAM-1 and VCAM-1 was downregulated by WEP treatment in a dose-dependent manner. Between the groups of 100 and 200 mg/L WEP, the difference was significant. In the injured HUVECs treated with 50 mg/L WEP, the inhibitory effect on the expression of ICAM-1 and VCAM-1 was presented in a time-dependent manner. Compared to the single administration, the use of WEP combined with fluvastatin showed better inhibitory effect on the expression of ICAM-1 and VCAM-1 in the injured HUVECs induced by TNF-α (P<0.01).CONCLUSION: WEP may be helpful for the protection of vascular endothelial cells by inhibiting the expression of ICAM-1 and VCAM-1 in a time-and dose-dependent manner. The protective effect of WEP on endothelial cells may be synergic with the inhibitor of HMG-CoA reductase such as fluvastatin sodium.     

he effects of constant magnetic field on apoptosis,adhesion molecule expression in endothelial cells and their adhesion rates with monocytes

AIM: To investigate the effects of constant magnetic field on apoptosis, secretion and expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVEC), and their adhesion rates with THP-1 monocytes induced by angiotensin Ⅱ (AngⅡ).METHODS: The third passage of cultured HUVEC was used.There were six groups: control group, Ang Ⅱ (10^-6 mol/L) group, Ang Ⅱ with 1, 5, 10 or 20 gausses of constant magnetic field group.Samples were collected 24 h after incubation with or without magnetic field.Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and propidinm iodide staining with flow cytometry.Secretion and expression of ICAM-1 and VCAM-1 were detected by ELISA and immunocytochemistry, respectively.Adhesion rate between HUVEC and THP-1 was measured by counting method.RESULTS: Ang Ⅱ at concentration of 10^-6mol/L induced apoptosis in HUVECs (P<0.05 vs control), whereas in 1, 5, 10 and 20 gausses group, apoptosis of HUVECs was significantly lower than that in Ang Ⅱ group (P<0.05).Ang Ⅱ at concentration of 10-6 mol/L significantly increased secretion and expression of ICAM-1 and VCAM-1 (P<0.05 vs control), whereas secretion and expression of ICAM-1 and VCAM-1 in 1, 5, 10 and 20 gausses group significantly decreased, compared with Ang Ⅱ group (P<0.05).The adhesion rates between HUVEC and THP-1 significantly increased 24 h after incubation of HUVEC with Ang Ⅱ (P<0.05 vs control), in contrast, the adhesion rates between HUVEC and THP-1 in 1, 5, 10 and 20 gausses group significantly decreased, compaed with Ang Ⅱ group (P<0.05).CONCLUSIONS: One gauss to 20 gausses of constant magnetic field antagonizes the effects of Ang Ⅱ on HUVEC, decreases apoptosis and expression of ICAM-1 and VCAM-1 in HUVEC, and also decreases the adhesion rates between HUVEC and monocytes induced by Ang Ⅱ.     

Dual roles of oxidized LDL in modulating expression of inflammatory molecules in HUVECs

AIM:To determine the role of LOX-1/PPAR pathway in regulating expression of adhesion molecules elicited by oxidizing low density lipoprotein(Ox-LDL) through Lectin-like oxidized low-density lipoprotein receptor-1(LOX-1) in human umbilical vein endothelial cells(HUVECs). METHODS:HUVECs were incubated with Ox-LDL,poly(I),carrageenan or 15-deoxy-△12,14-prostaglanding J2 (15d-PGJ₂ ). PPAR mRNA and protein were examined by real time RT-PCR and Western blotting. ICAM-1 and E-selectin were detected by RT-PCR and Western blotting respectively. RESULTS:Ox-LDL increased PPAR expression in HUVECs,which was inhibited by pretreatment of HUVECs with LOX-1 blockers. Preincubation of HUVECs with 15d-PGJ₂ attenuated the expression of intercellular adhesion molecule-1(ICAM-1) and E-selectin in response to Ox-LDL. Upregulation of ICAM-1 and E-selectin mediated by Ox-LDL were suppressed more significantly by the combination of 15d-PGJ₂ and polyinosonic acid as compared to either 15d-PGJ₂ or polyinosonic acid alone. CONCLUSION:The results indicate that Ox-LDL exerts a biphasic effects on inflammatory response. It evokes harmful effects by inflammatory injury on one side and protective effects by triggering the LOX-1/ PPAR signaling pathway on the other hand.     

Effects of xijiao dihuang decoction on the expression of adhesion molecules in vascular endothelial cells of adrenine and hypothermia-treated rats

AIM: To study the effects of xijiao dihuang decoction, a Chinese medicine, on the expression of adhesion molecules in vascular endothelial cells (VECs) of adrenine and hypothermia-treated rats. METHODS: The expression of VCAM-1, ICAM-1, PECAM-1, iNOS and their corresponding mRNA in VECs was assayed by immunohistochemistry analysis and RT-PCR, respectively. RESULTS: Immunohistochemistry analysis showed the expression of VCAM-1, ICAM-1, PECAM-1 and iNOS are higher in adrenine and hypothermia-treated rats than that in controls, xijiao dihuang decoction decreased the expression of VCAM-1, ICAM-1, PECAM-1 and iNOS in a dose-dependent manner in adrenine and hypothermia-treated rats. CONCLUSION: Xijiao dihuang decoction can down-regulate the expression of adhesion molecules in vascular endothelial cells of adrenine and hypothermia-treated rats.     

Limb ischemic preconditioning protects human umbilical vein endothelial cells from oxidative stress induced by H2O2

AIM: To investigate the effects of the sera from the rats after limb ischemic preconditioning (LIPC) on human umbilical vein endothelial cells (HUVECs) injured by hydrogen peroxide (H₂O₂). METHODS: The HUVECs were divided into 5 groups: the cells in control group were cultured without any intervention; the cells in model group (M) were damaged by 1 mmol/L H₂O₂ for 2 h; the cells in early preconditioning serum (EPS) group, delayed preconditioning serum (DPS) group or sham limb ischemic preconditioning serum (SPS) group were treated with the corresponding serum at 5% for 12 h, respectively, and then treaed with H₂O₂ for 2 h. The viability of the HUVECs was analyzed by flow cytometry. The lactate dehydrogenase (LDH) in the culture media was detected. The cell adhesion molecules in the HUVECs were detected by real-time PCR. The mRNA and protein expression of heme oxygenase-1 (HO-1) was also determined. RESULTS: The viability of HUVECs incubated with 1 mmol/L H₂O₂ for 2 h significantly decreased compared with the control cells, which was accompanied with the augmentations of LDH in the medium and the cell adhesion molecules in cells, such as vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1). Preincubation with EPS and DPS derived from the rats subjected LIPC attenuated these injuries. Furthermore, pretreatment with EPS and DPS increased the expression of HO-1 at mRNA and protein levels. CONCLUSION: LIPC protects the HUVECs from H₂O₂-induced injury.     

Effects of Xijiao Dihuang and Yinqiao San decoction on ICAM-1 and VCAM-1 expression in mouse lung tissues and rat pulmonary microvascular endothelial cells infected with influenza virus

AIM：To investigate the effects of Xijiao Dihuang and Yinqiao San decoction (XDY) on the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in mouse lung tissues and rat pulmonary microvascular endothelial cells (RPMVECs) infected with influenza virus, and to explore its mechanism for treatment of viral pneumonia. METHODS：Fifty-four male BALB/c mice were randomly divided into normal group, model group and XDY group (n=18 in each group). The viral pneumonia model was established by intranasally dripping influenza A (H1N1) virus into the mice. The mice in XDY group were treated with XDY 1 h after dripping the virus. The expression of ICAM-1 and VCAM-1 in lung tissues was examined by immunohistochemical staining 2, 4 and 6 d after infection. On the other hand, RPMVECs were obtained from male Wistar rats and primarily cultured. The cells were randomly divided into control group, virus group, virus+XDY group, tumor necrosis factor α (TNF-α) group and TNF-α+XDY group. The mRNA and protein expression of ICAM-1 and VCAM-1 was evaluated by real-time PCR and flow cytometry 24 h after infection. RESULTS：Virus exposure increased ICAM-1 and VCAM-1 expression in mouse lung tissues (P<0.01), and XDY treatment attenuated this effect (P<0.01). Virus and TNF-α both led to the increases in mRNA and protein expression of ICAM-1 and VCAM-1 in RPMVECs (P<0.01), which were also reduced by treatment with XDY (P<0.01). CONCLUSION：Treatment with XDY decreases virus-induced ICAM-1 and VCAM-1 expression, suggesting an important role of XDY in treatment of viral pneumonia.     

Effects of dehydroepiandrosterone on expression of ICAM-1 in rabbit aorta and HUVECs induced by high lipid levels

AIM: To investigate the effects of dehydroepiandrosterone (DHEA) on the expression of intercellular adhesion molecule-1 (ICAM-1) induced by high lipid levels in rabbit aorta and human umbilical venous endothelial cells (HUVECs), and the effects of all-trans retinoic acid (ATRA) in this process.METHODS: For in vitro experiments, the cultured HUVEC were divided into control group, oxidized low-density lipoprotein (ox-LDL) group, ox-LDL+DHEA group, ox-LDL+DHEA+ATRA group and DHEA group. The HUVECs in all groups were treated with the corresponding reagents for 24 h. The expression of ICAM-1 at mRNA and protein levels in all groups were determined by RT-PCR and ELISA, respectively. For in vivo experiments, the rabbits were divided into control group, high lipid group, high lipid+DHEA group, high lipid+DHEA+ATRA group and DHEA group. The rabbits in all groups were fed with the corresponding diets for 10 weeks. The expression of ICAM-1 in the rabbit aorta at mRNA and protein levels was determined by RT-PCR and immunohistochemistry. RESULTS: The expression of ICAM-1 in the HUVECs in ox-LDL group was significantly increased compared with control group (P<0.05). Compared with ox-LDL group, the expression of ICAM-1 in ox-LDL+DHEA group was obviously decreased (P<0.05). The expression of ICAM-1 was similar in both control group and DHEA group (P>0.05). The expression of ICAM-1 was similar in both ox-LDL+DHEA group and ox-LDL+DHEA+ATRA group (P>0.05). The expression of ICAM-1 in the rabbit aorta in high lipid group was significantly increased compared with control group (P<0.05). Compared with high lipid group, the expression of ICAM-1 in high lipid+DHEA group was obviously decreased (P<0.05). No remarkable difference in the expression of ICAM-1 between control group and DHEA group was observed (P>0.05), so did between high lipid+DHEA group and high lipid+DHEA+ATRA group (P>0.05). CONCLUSION: DHEA inhibits high lipid-induced ICAM-1 expression in rabbit aorta and HUVECs. That may be one of the mechanisms of antiatherosclerotic effect of DHEA. ATRA seems no positive effect on DHEA function.     

Effect of vitamin E on ox-LDL-induced monocyte/endothelium adhesion

AIM:To observe the role of ox-LDL in rabbit endothelium/circulating monocyte adhesion in vitro and the effect of vitamin E.METHODS:Cultured rabbit's endothelial cells were incubated with different concentrations of ox-LDL, then incubated with rabbit's monocytes to observe cell's adhesion. With Northern blotting, endothelial VCAM-1 mRNA expression was measured. By using vitamin E incubation before ox-LDL, above steps were repeated to observe the effect of vitamin E.RESULTS:When ox-LDL concentrations were 2.5 mg/L,5 mg/L,10 mg/L,monocytes adhesive to endothelium per microscope field were 132.8±20.2,350.0±37.2,502.6±78.8,respectively They were al significantly higher than that of control group(51.2±7.7,P<0.01)Addition of vitamin Eat comcentration of 10 mol/L,20 mol/L,40 mol/L respect ively before ox-LDL incubation yield fewer adhesive monocytes as follow:422.3±32.2,296.0±21.7,205.2±36.6,respectively,but only the last two concentrations had significant effects(P<0.01)Endothelial VCAM-1 mRNA expression in ox-LDL group was higher than that in vitamin E group(0.49±0.09 vs 0,33±0.10,P<0.05).CONCLUSIONS:ox-LDL directly enhances rabbit's endothelium/monocyte adhesion by inducing expression of VCAM-1 in endothelium. Vitamin E inhibites these effects of ox-LDL.     

Chlamydia pneumoniae enhanced the expression of ICAM-1 in cultured human umbilical vein endothelial cells

AIM: To investigated the effects of Chlamydia pneumoniae (C.pneumoniae) infection on the expression of intercellular adhesion molecule-1 (ICAM-1) in cultured human umbilical vein endothelial cells (HUVECs). METHODS: After propagated in HEP-2 cells, C. pneumoniae organisms were infected to HUVECs. The infection was assessed by ectromicroscope and PCR. The expression of ICAM-1 on HUVECs was detected by flow cytometry before infection and 12 h, 24 h, 48 h, 72 h after infection. RT-PCR was used to detect the ICAM-1 mRNA expression. RESULTS: C.pneumoniae was able to infect cultured HUVECs. After infection, the expression of ICAM-1 on the surface of cultured HUVECs was increased,, the peak was at the time of about 24-48 h; The light quantative RT-PCR analysis demonstrated that the infection also enhanced the ICAM-1 mRNA expression. CONCLUSION: The ability of C.pneumoniae to grow in HUVECs and to stimulate the expression of ICAM-1 protein and mRNA suggests that C.pneumoniae may playa role in atheriosclerosis.     

Effects of oxidized HDL on the levels of MCP-1, ICAM-1 and free calcium in cultured human umbilical venous endothelial cells

AIM:To study the effects of oxidized high-density lipoprotein (oxHDL) on the expression of monocyte chemoattractant protein-1(MCP-1) and intercellular adhesion molecule-1(ICAM-1) and intracellular free calcium concentration ([Ca²⁺]i) level in cultured human umbilical venous endothelial cells(HUVECs). METHODS:The MCP-1 protein content in the medium of conditioned HUVEC was measured by ELISA, and the ICAM-1 on HUVECs was detected by indirect immunofluorescence, and [Ca²⁺]i was determined by Fluo-3/AM, the injury of cells was observed by scanning electron microscopy (SEM).RESULTS:oxHDL could induce the expression of MCP-1 and ICAM-1 in HUVECs. In oxHDL group (HUVECs were incubated with 100 mg protein/L oxHDL for 24 h), the levels of MCP-1, ICAM-1 and [Ca²⁺]i increased by 160%, 60% and 70% respectively compared with the control group (P＜0.01). When HUVECs were incubated with 300 mg protein/L oxHDL for 24 h, cells were injured obviously. CONCLUSION:By inducing the expression of ICAM-1 and MCP-1 in endothelial cells, oxHDL may promote monocyte-endothelium adhesion and monocyte migration to intima, it may promote atherosclerosis as oxidized low-density lipoprotein (oxLDL).     

Effects of Huaiyu decoctum on serum concentrations of TNF-α, IL-6 and IL-10 in rats after anorectal operation

ATM: To investigate the effects of Huaiyu decoctum on the serum concentrations of TNF-α, IL-6 and IL-10 in rats after anorectal operation. METHODS: Sprague-Dawley rats (n=40) were randomly divided into 4 groups:normal group, model group, low-dose Huaiyu decoctum group and high-dose Huaiyu decoctum group. The concentrations of TNF-α, IL-6 and IL-10 in the rat serum were measured by ELISA. The pathologic changes of the anorectal tissues were observed under microscope with HE staining. The protein expression of ICAM-1, VCAM-1 and NF-κB was determined by Western blotting. RESULTS: After Huaiyu decoctum administration, TNF-α and IL-6 concentrations in the serum were significantly decreased, and IL-10 concentration was increased as compared with model group. Moreover, Huaiyu decoctum markedly attenuated edema and hyperemia in the rats after anorectal operation. The protein expression of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and NF-κB in the anorectal tissues was obviously inhibited by Huaiyu decoctum treatment. CONCLUSION: Huaiyu decoctum improves the recovery of anorectal tissues after operation by decreasing the serum concentrations of TNF-α, IL-6 and IL-10, and reducing the protein expression of ICAM-1, VCAM-1 and NF-κB in the anorectal tissues.     

NF-κB site of promotor mediates nicotine-induced ICAM-1 expression in human umbilical vein endothelial cells

AIM:To study the mechanisms of nicotine-induced expression of intercellular adhesion molecule-1(ICAM-1).METHODS:Related luciferase reporter gene plasmids were constructed with molecular cloning techniques;above plasmids and intracontrol plasmid pSV-β-gal were co-transfected into human umbilical vein endothelial cells(HUVECs) with eukaryotic gene transfection techniques; the relative luciferase activities were detected in the transfected HUVECs.RESULTS:Series of luciferase reporter gene containing different sequences of human ICAM-1 promotor and site-directed mutants of NF-κB and Sp-1 in promotor were successfully constructed; Nicotine could increase the expression of luciferase reporter gene plasmid containing-579 bp(pGL3E-579/+36),-230 bp(pGL3E-230/+36) and mutated Sp-1 version(pGL3E-Sp-1-MU)(P＜0.05 vs control) of ICAM-1 promotor in the transfected HUVECs, whereas deletion derivative (pGL3E-134/+36) and mutation (pGL3E- NF-κB -MU) of downstream NF-κB site of ICAM-1 promotor prevent nicotine-induced increase in expression of luciferase reporter gene plasmid.CONCLUSION:NF-κB site of promotor mediates nicotine-induced ICAM-1 expression in human umbilical vein endothelial cells.     

Role of MEK1/2 in ICAM-1 over expression induced by lipopolysaccharide in human umbilical vein endothelial cells

AIM: To investigate the role of MEK1/2, a subfamily of mitogen activated protein kinase-kinase (MAPKK), in expression of intercellular adhesion molecule-1 (ICAM-1) induced by lipopolysaccharide (LPS) in human umbilical vein endothelial cells (HUVECs). METHODS: Expression levels of ICAM-1 mRNA and its protein were assayed using RT-PCR and Western blotting, respectively, in HUVECs pretreated with different concentrations of LPS for different times with or without PD98059, a specific inhibitor of MEK1/2. RESULTS: LPS up-regulated the expression of ICAM-1 mRNA and its protein in HUVECs in a concentration- and time-dependent manners. The expression levels of ICAM-1 mRNA and its protein began to elevate at 2 h after LPS treatment, and reached nearly a peak value at 6 h after LPS (100 μg·L-1) treatment. PD98059 (10 μg·L-1) significantly inhibited LPS-induced expression of ICAM-1 mRNA and protein, the expression inhibitory rates of which were 54.4% and 44.9%, respectively (P<0.01). CONCLUSION: Modulation of MEK1/2 signaling pathway might be a new and useful strategy for the prevention and treatment of vascular endothelial injury induced by LPS.     

Effect of N,N-dimethylsphingosine on the expression of adhesion molecules in human umbilical vein endothelial cell line EA.hy926

AIM: To investigate whether and how N, N-dimethylsphingosine (DMS) plays a role in modulating the adhesion of monocytes to vascular endothelial cells, and identify whether human umbilical vein endothelial cell line EA.hy926 take place of the vascular endothelial cells.METHODS: Adhesion ratio was measured by flow cytometry, and immunohistochemistry was used to detect the expression of ICAM-1 and P-selectin in HUVEC: EA.hy926 cells after the effect of DMS. RESULTS: DMS inhibited the adhesion of monocytes to HUVEC: EA.hy926 cells in a time-dependent and concentration-dependent manner by reducing the expression of ICAM-1 and P-selectin. CONCLUSIONS: DMS reduced adhesion molecule expression in vascular endothelial cells. DMS may be an important contributor to reduce adhesion ratio, suggesting that DMS plays a negative role in proinflammatory and immune functions of the modified vascular endothelial cells during atherosclerosis and restenosis.     

Effects of Astragalus polysacharin on fibroblast proliferation and adhesion between HUVECs and white cells

AIM: To investigate the effects of Astragalus polysacharin(APS) on human fibroblast and human umbilical vein endothelia cell (HUVEC) proliferation, as well as its acts on adhesion between white cells and HUVECs. METHODS: Human fibroblasts from distal and proximal skin away the ulcer were cultured as normal fibroblasts(NF) and wounded fibroblasts(WF). MTT assay was used for detecting cell proliferation, Rose Bengal staining and fluorescence immunohistology assay were used for examining the adhesion of human polymorpho-nuclear cell(PMN) and TPH-1 to HUVECs. RESULTS: 2.44-156 mg/L APS promoted WF proliferation, and 2.44-39 mg/L APS also promoted NF proliferation, but it did not show any proliferating effect on HUVECs. APS inhibited the adhesion of PMN or TPH-1 to HUVECs induced by tumor necrosis factor(TNF). At 25-100 mg/L, it also inhibited both VCAM-1 and ICAM-1 expression in HUVECs induced by TNF. Treatment with APS for 12 h also inhibited CD44 expression in HUVECs. CONCLUSION: APS shows mitogenic activity on both human normal and wounded fibroblasts. It also exerts anti-inflammation effects by inhibiting adhesion molecule expression and adhesion of white cells to HUVECs.     

Inhibition of ICAM-1 expression on endothelial cells in hypoxia/reoxygenation by antisense oligodeoxynucleotides

AIM: To investigate the effect of antisense oligodeoxynucleotides (AS-ODN) on the intercellsular adhesion molecule-1(ICAM-1) expression on endothelial cells in hypoxia/reoxygenation(H/R). METHODS: With cultured glomerular vascular endothelial cell in H/R, the positive percentage of ICAM-1 expression was measured by flow cytometry before and after giving AS-ODN. RESULTS: The ICAM-1 expression did not increase on glomerular vascular endothelial cell in 10 hours hypoxia compared to control group, it increased in 6 hours reoxygenation, and decreased by 40.6% after giving AS-OND. CONCLUSION: AS-ODN may decrease the expression of ICAM-1 on endothelial cells in H/R.     

Effect of pregnancy-associated glycoproteins on E-selectin-medi ated endothelial cell adhesion

AIM:To explore the effect of high purified human chorionic gonadotropin (hCGs) and glycodelins isolated from amniotic flui d and serum of pregnant women on E-selectin mediated endothelial cell adhesion.METHODS:The hCGs and glycodelins in human amniotic fluid and se rum collected from pregnant women were purified by using chromatography.The bin ding of[51Cr］-labelled HepG2 cells to IL-1β stimutated human umbilical vein endothelial cells (HUVECs) in the presence of increasing concentrations of hCGs and glycodelins was quantified.The effect of hCG and glycodelin on E-selec tin-mediated endothelial cell adhesion was observed.RESULTS:HCGs isolated from amniotic fluid and serum of pregnant women inhibited the E-selectin mediated adhesion of HepG2 cells to HUVECs in a dose-dependent manner,the IC₅₀ was 3.2×10-6 mol/L and 6.3×10 -6 mol/L,respectively and the relative inhibition was 1.5×104 and 1 .1×104,respectively,compared to sialyl-Lewisx.The IC₅₀ of amnioti c fluid glycodelin and serum glycodelin was 3.2×10-6 mol/L and 6.3×10 -6 mol/L respectively,and the relative inhibition was 7.2×102 and 3.7×102,respectively,compared with sialyl-Lewisx.CONCLUSION:Pregnancy-associated glycoproteins,hCG and glycodel in,are the powerful inhibitors for E-selectin-mediated endothelial cell adhesion.