2.Isolation and pure culture of microvascular endothelial cells from the fetal skin.
Myung Soo CHA ; Dong Kyun RAH ; Kwang Hoon LEE
Yonsei Medical Journal 1996;37(3):186-193
Microvascular endothelial cells were purely isolated from human fetal skin using magnetic particles. The principle of this technique is based on the selective binding of the lectin Ulex europaeus I (UEA I) to the endothelial cell surface via fucose residues. Initially UEA I was covalently bound to tosyl-activated magnetic polydisperse polymer particles (Dynabeads) and then the UEA I-coated beads were collected using a magnetic particle concentrator (MPC). Endothelial cells were isolated by extracting microvascular segments from trypsin-treated fetal skin tissue and were purified by sieving with nylon mesh and by 35% Percoll gradient centrifugation. For further purification, the obtained cells were incubated with UEA I-coated Dynabeads. The endothelial cells bound to the Dynabeads were collected using MPC. This is a simple and reproducible technique for isolating a pure population of microvascular endothelium from the fetal skin.
Cells, Cultured
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Endothelium, Vascular/*cytology
;
Factor VIII/analysis
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Female
;
Fetus
;
Human
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Intercellular Adhesion Molecule-1/analysis
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Pregnancy
;
Skin/blood supply
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Support, Non-U.S. Gov't
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Vascular Cell Adhesion Molecule-1/analysis
3.Study on anti-angiogenesis effect of three curcumin pigments and expression of their relevant factors.
Yan-fen HUANG ; Xue-xin ZHU ; Zhi-shan DING ; Gui-yuan LV
China Journal of Chinese Materia Medica 2015;40(2):324-329
To study the in vitro anti-angiogenesis effect of three curcumin pigments (curcumin, demethoxycurcumin, bisdemethoxycurcumin). In the study, the inhibitory effect of the three curcumin pigments on proliferation of HUVEC cells induced by OX-LDL and the effect on migration of HUVEC cells were detected. The effect on neovascularization was observed by chorioallantoic membrane (CAM) test. The effect on cell adhesion factors ICAM-1 and VCAM-1 of HUVECs were tested by Real-time RT-PCR. It was found that the three curcumins could inhibit the proliferation of HUVEC cells induced by OX-LDL within the dosage range 4, 8, 16 mg x L(-1), with a dose-dependence. The proliferative effect of curcumins on HUVECs was greater than the other two derivatives (P < 0.01). All of the three curcumin pigments inhibited the migration of HUVEC cells and the angiogenesis of chick chorioallantoic membrane (CAM). The migration inhibition rate of curcumins at middle and high concentrations was greater than the other two (P < 0.01). All of the three curcumin could down-regulate the expression of VEGF and ICAM-1, and curcumins showed more obvious effect in down-regulating VEGF than demethoxycurcumin and bisdemethoxycurcumin(P < 0.01); Bisdemethoxycurcumin showed the most significant effect in down-regulating ICAM-1 (P < 0.01). All of the three showed no remarkable effect on expression of VCAM-1, and only bisdemethoxycurcumin showed the down-regulating effect (P < 0.05). According to the findings, all of the three curcumin pigments could resist angiogenesis by inhibiting proliferation and migration of endothelial cells and down-regulating the expression of VEGF and adhesion molecules ICAM-1.
Angiogenesis Inhibitors
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pharmacology
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Animals
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Cell Movement
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drug effects
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Cells, Cultured
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Chorioallantoic Membrane
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drug effects
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Curcumin
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analogs & derivatives
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pharmacology
;
Humans
;
Intercellular Adhesion Molecule-1
;
genetics
;
RNA, Messenger
;
analysis
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Vascular Cell Adhesion Molecule-1
;
genetics
;
Vascular Endothelial Growth Factor A
;
genetics
4.Effect of cytokines on the expression of cell adhesion molecule and on the adhesion of melanoma cells to endothelial cells.
Se Jong KIM ; Nam Soo KIM ; Jung Lim LEE
Journal of Korean Medical Science 1993;8(1):41-52
We examined the role of cell adhesion molecules (CAM) by which tumor cells bind to the endothelial cells using human umbilical vein endothelial cells (HUVEC) and cultured melanoma cells. Endothelial cells from human umbilical veins were isolated and examined for CAM expression and its modulation by tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6) or interferon-gamma (IFN-gamma). The expression of intercellular adhesion molecule 1 (ICAM-1) on HUVEC was increased by TNF-alpha, IL-1 and IFN-gamma when measured by ELISA or flow cytometric (FACS) analysis. IL-6 did not increase ICAM-1 expression on HUVEC. Two melanoma cell lines, Malme-3M and SK-Mel-28, showed increased expression of ICAM-1 after treatment with TNF-alpha, IL-1 and IFN-gamma in FACS analysis. IFN-gamma induced increased expression of HLA-DR only in SK-Mel-28 melanoma cells, not in Malme-3M melanoma cells. Neither HUVEC nor melanoma cells expressed lymphocyte function-associated antigen 1 (LFA-1) in either the basal (i.e., cytokine untreated) condition or the cytokine treated condition. Melanoma cells showed minimal increment in adhesion to TNF-alpha or IL-1 treated HUVEC than to cytokine untreated HUVEC. HUVEC and melanoma cells did not express LFA-1 and increased ICAM-1 expression by TNF-alpha, IL-1 and IFN-gamma treatment in FACS analysis did not coincide with minimal increase of melanoma cells adhesion to cytokine treated HUVEC. These results suggest that adhesion between melanoma cells and HUVEC is probably mediated by molecular interaction other than ICAM-1/LFA-1.
Cell Adhesion/drug effects
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Cell Adhesion Molecules/*analysis
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Cell Division/drug effects
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Cells, Cultured
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Cytokines/*pharmacology
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Endothelium, Vascular/cytology/*physiology
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HLA-DR Antigens/analysis
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Humans
;
Intercellular Adhesion Molecule-1
;
Lymphocyte Function-Associated Antigen-1/analysis
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Melanoma/*pathology
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Tumor Cells, Cultured
5.Experimental study on mechanism and rarity of metastases in skeletal muscle.
Chenghua LUO ; Yanyong JIANG ; Yongxue LIU ; Xianghong LI
Chinese Medical Journal 2002;115(11):1645-1649
OBJECTIVETo investigate the reasons for the rarity of metastases in skeletal muscle.
METHODSBy injecting tumor cells (Walker256 rat carcinosarcoma) through the iliac artery (experimental group) and the tail vein (control group), animal models of blood-borne metastases were established. The quadriceps femoris muscle and lungs were observed grossly and microscopically. Immunohistochemistry was applied to investigate the expression of vascular cell adhesion molecule-1 (VCAM-1) in the microvascular endothelium of these organs. Primary culture of rat skeletal muscle cells was established and conditioned medium (MCM) was collected. Effects of MCM on several tumor cell lines and the biochemical characteristics of skeletal muscle delivered tumor factor(s) were tested by MTT assay. Apoptosis and morphological examination were carried out to investigate the antitumor mechanisms of MCM.
RESULTSIn the experimental group, there were no definite metastases observed in muscle cells. In the control group, lung metastases were present in the lungs of all rats that were sacrificed at the 14th day or died spontaneously (17 rats in all). There was no significant difference between the increase in VCAM-1 in quadriceps femoris muscle 7 days after iliac artery injection and that in lungs 7 days after tail vein injection (P > 0.05). In vitro studies showed that the proliferation of tumor cell lines of mouse SP2/0 myeloma, rat Walker256 carcinosarcoma or human chronic granulocytic leukemia K562, human acute lymphatic leukemia HL-60, LS-174-T colon adenocarcinoma, PC3-M prostatic carcinoma and lung giant cell carcinoma with different metastatic potency (PLA801-C with low metastatic potency, PLA801-D with high metastatic potency) was significantly inhibited when cultured with MCM (P < 0.01 - 0.05). Proliferation of malignant cells showed a dose-dependent decrease, to a certain degree. Proliferation of normal rabbit joint epiphysial disk cells (RGP-2) were not affected by MCM. Proliferation of lung giant cell carcinoma cells with high metastatic potency showed a significant decrease even when cultured in highly diluted MCM (6.25% of primary MCM), when compared with the strain of low metastatic potency. Following ultrafiltration, boiling at 100 degrees C, and treatment with trypsin, skeletal muscle delivered tumor factor(s) were found to be a low molecular weight (MW CONCLUSIONSThe rarity of metastases in skeletal muscles, generally accepted in the clinical setting, can be reproduced in an animal model. It does not seem to be related to VCAM-1 expression in the microvessels of these organs. Skeletal muscle delivered factor(s) play a key role in the mechanism of the rarity of metastases in skeletal muscle.
Animals
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Cell Division
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Humans
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Immunohistochemistry
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Muscle Neoplasms
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pathology
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secondary
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Muscle, Skeletal
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physiology
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Rats
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Rats, Wistar
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Tumor Cells, Cultured
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Vascular Cell Adhesion Molecule-1
;
analysis
6.Heparin-derived oligosaccharide inhibits vascular intimal hyperplasia in balloon-injured carotid artery.
Jie-Ru LIU ; Jie WU ; Xin-Chao YU ; Xuan QIAN ; Rui XIONG ; Hui-Fang WANG ; Dan-Feng YU ; Fei-Fei LIU ; Shu-Ying HE
Chinese Journal of Natural Medicines (English Ed.) 2017;15(6):442-450
The aims of the present study were to determine the effects of heparin-derived oligosaccharides (HDOs) on vascular intimal hyperplasia (IH) in balloon-injured carotid artery and to elucidate the underlying mechanisms of action. An animal model was established by rubbing the endothelia within the common carotid artery (CCA) in male rabbits. The rabbits were fed a high-cholesterol diet. Arterial IH was determined by histopathological changes to the CCA. Serum lipids were detected using an automated biochemical analysis. Expressions of mRNAs for vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), vascular cell adhesion molecule-1 (VCAM-1), monocyte chemoattractant protein-1 (MCP-1), scavenger receptor class B type I (SR-BI), and ATP-binding cassette transporter A1 (ABCA-1) were analyzed using reverse transcription polymerase chain reaction assays. Expressions of VEGF, VCAM-1, MCP-1, SR-BI and ABCA-1 proteins were analyzed by Western blotting. Enzyme-linked immunosorbent assays were used to quantify expression levels of VEGF and bFGF. Our results showed that administration of HDO significantly inhibited CCA histopathology and restenosis induced by balloon injury. The treatment with HDOs significantly decreased the mRNA and protein expression levels of VEGF, bFGF, VCAM-1, MCP-1, and SR-BI in the arterial wall; however, ABCA-1 expression level was elevated. HDO treatment led to a reduction in serum lipids (total cholesterol, triglycerides, high-density and low-density lipoproteins). Our results from the rabbit model indicated that HDOs could ameliorate IH and underlying mechanism might involve VEGF, bFGF, VCAM-1, MCP-1, SR-BI, and ABCA-1.
ATP Binding Cassette Transporter 1
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analysis
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Animals
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Carotid Artery Injuries
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drug therapy
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pathology
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Chemokine CCL2
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analysis
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Heparin
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therapeutic use
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Hyperplasia
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Male
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Oligosaccharides
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therapeutic use
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Rabbits
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Tunica Intima
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pathology
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Vascular Cell Adhesion Molecule-1
;
analysis
;
Vascular Endothelial Growth Factor A
;
analysis
7.Expression of vascular cell adhesion molecule-1 in lung slices from antigen sensitized rats and pharmacological modulation.
Journal of Zhejiang University. Medical sciences 2003;32(4):319-322
OBJECTIVETo investigate the expression of vascular cell adhesion molecule-1(VCAM-1) in lung slices from antigen -sensitized rats and the modulation by drugs.
METHODSIn isolated lung slices from ovalbumin(OVA)-sensitized rats, the relative expression of VCAM-1 was determined after drug treatment and OVA challenge.
RESULTThe expression of VCAM-1 was enhanced in the sensitized rat lungs,and OVA challenge did not further increase the expression. Glycocorticosteroid dexamethasone and leukotriene cysLT receptor antagonist ONO-1078 inhibited the expression,but tachykinin NK-1 receptor antagonist SR-140333 had no such effect.
CONCLUSIONVCAM-1 expression is enhanced in the sensitized rat lungs, and antigen challenge does not further up regulate the expression. Anti-inflammatory drugs have different effects on VCAM-1 expression. Dexamethasone and ONO-1078, but not SR-140333, can inhibit the expression.
Animals ; Chromones ; pharmacology ; Dexamethasone ; pharmacology ; Female ; In Vitro Techniques ; Lung ; chemistry ; Male ; Ovalbumin ; immunology ; Piperidines ; pharmacology ; Quinuclidines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Vascular Cell Adhesion Molecule-1 ; analysis
8.Expression of cell adhesion molecules in acute leukemia cell.
Xiaoping JU ; Min PENG ; Xiaoping XU ; Shuqing LU ; Yao LI ; Kang YING ; Yi XIE ; Yumin MAO ; Fang XIA
Chinese Journal of Hematology 2002;23(11):581-584
OBJECTIVETo investigate the role of cell adhesion molecule in the development and extramedullary infiltration (EI) of acute leukemia.
METHODSThe expressions of neural cell adhesion molecule (NCAM) gene, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule (VCAM-1) genes in 25 acute leukemia patients bone marrow cells were detected by microarray and reverse transcriptase-polymerase chain reaction (RT-PCR).
RESULTSThe expressions of NCAM, ICAM-1 and VCAM-1 gene were significantly higher in acute leukemia cells and leukemia cells with EI than in normal tissues and leukemia cells without EI, respectively, both by cDNA microarray and by RT-PCR.
CONCLUSIONThe cDNA microarray is a powerful technique in analysis of acute leukemia cells associated genes. High expressions of cell adhesion molecule genes might be correlated with leukemia pathogenesis and infiltration of acute leukemia cell.
Acute Disease ; Adolescent ; Adult ; Aged ; Bone Marrow Cells ; metabolism ; pathology ; Cell Adhesion Molecules ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Leukemia, Myeloid ; genetics ; pathology ; Male ; Middle Aged ; Neural Cell Adhesion Molecules ; genetics ; Oligonucleotide Array Sequence Analysis ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Cell Adhesion Molecule-1 ; genetics
9.Polysaccharide sulfate 916 inhibits neutrophil-endothelial adhesion.
Decheng REN ; Meiyu GENG ; Guanhua DU ; Juntian ZHANG
Chinese Medical Journal 2002;115(12):1855-1858
OBJECTIVETo study the effect of polysaccharide sulfate 916 (PS916) on neutrophil-endothelial cell adhesion.
METHODSCell 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.
RESULTSTumor necrosis factor alpha (TNFalpha, 50 - 800 U/ml) increased the adherence of neutrophil to TNFalpha-stimulated HUVEC in a concentration and time dependent manner. PS916 (0.01 - 1.0 mg/ml) dose-dependently inhibited the adherence of neutrophils to TNFalpha-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 TNFalpha-stimulated HUVEC.
CONCLUSIONSPS916 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).
Animals ; Cell Adhesion ; drug effects ; Cells, Cultured ; Endothelium, Vascular ; cytology ; Humans ; Intercellular Adhesion Molecule-1 ; analysis ; N-Formylmethionine Leucyl-Phenylalanine ; pharmacology ; Neutrophils ; drug effects ; physiology ; Polysaccharides ; pharmacology ; Rats ; Rats, Wistar ; Sulfuric Acids ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology ; Vascular Cell Adhesion Molecule-1 ; analysis
10.Effects of retinoic acid on adhesion molecules expression and adhesive function in murine bone marrow stromal cell.
Hui-Rong MAI ; Shao-Liang HUANG ; Jing WEI ; Yan-Feng WU
Journal of Experimental Hematology 2002;10(6):499-502
To explore the effect of retinoic acid (RA) on the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in murine bone marrow stromal cell (BMSC) and adhesive rate of human cord blood mononuclear cell (UCBMNC) to BMSC in vitro, the express ions of ICAM-1 and VCAM-1 of murine BMSC were detected by flow cytometry and the binding capacity of UCBMNC to BMSC was tested by MTT assay after co-culturing with 0.1, 1.0 and 10.0 micro mol/L RA, respectively. The results showed that 1.0 and 10.0 micro mol/L RA increased the expression of ICAM-1 and the adhesive rate of U CBMNC to BMSC, however, RA did not induced the increase of expression of VCAM-1. It was positive correlation between the increments of ICAM-1 expression and the adhesive rate (r = 0.7883, P < 0.05). It is concluded that RA up-regulated ICAM-1 expression of BMSC and increased the adhesion of UCBMNC to BMSC in vitro. These may clarify the correlation between adhesion molecules on BMSC and homing of hematopoietic stem cells, and provide the experimental basis for RA to promote the homing of umbilical cord blood stem cell.
Animals
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Bone Marrow Cells
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chemistry
;
drug effects
;
physiology
;
Cell Adhesion
;
drug effects
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Fetal Blood
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cytology
;
Hematopoietic Stem Cells
;
physiology
;
Intercellular Adhesion Molecule-1
;
analysis
;
Leukocytes, Mononuclear
;
drug effects
;
physiology
;
Mice
;
Stromal Cells
;
chemistry
;
drug effects
;
physiology
;
Tretinoin
;
pharmacology
;
Vascular Cell Adhesion Molecule-1
;
analysis