[Effects of monocyte chemotactic protein-3 on ICAM-1, VCAM-1, TF, and TFPI expression and apoptosis in human umbilical vein endothelial cells].
OBJECTIVE
To investigate the effect of monocyte chemotactic protein-3 (MCP-3) on the expressions of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), tissue factor (TF, and tissue factor pathway inhibitor (TFPI) and cell apoptosis in human umbilical vein endothelial cells (HUVECs).


METHODS
Cultured HUVECs were treated with MCP-3 at the optimal concentration determined previously 1 h after treatments with or without MCP-3 antibody (20 ng/ml), PI3K inhibitor, or LY-294002 (5 mmol/ml). The expressions of ICAM-1, VCAM-1, TF and TFPI were analyzed using RT-PCR and Western blot after the treatments. MCP-3 mRNA and protein expressions were detected in HUVECs exposed to 50 µg/ml ox-LDL for 24 h. The cell apoptosis and caspase-3 protein production in HUVECs treated with MCP-3 or with MCP-3 plus CCR2 antagonist for 24 h and 48 h were evaluated by flow cytometry and Western blotting.


RESULTS
At the optimal concentration of 0.3 ng/ml, MCP-3 treatment for 24 h caused significantly increased ICAM-1, VCAM-1, and TF expressions with lowered expression of TFPI in HUVECs (P<0.05), and such effects were significantly inhibited by the application of MCP-3 antibody, PI3K inhibitor, or LY-294002 (P<0.05). Ox-LDL exposure significantly increased the expression of MCP-3 in HUVECs (P<0.05). HUVECs showed a significantly increased apoptosis rate after treatment with MCP-3 or with MCP-3 plus CCR2 antagonist (P<0.05), and the apoptosis rate increased significantly as the treatment time prolonged (P<0.05); caspase-3 protein expression in the cells showed a similar pattern of alterations following the treatments.


CONCLUSIONS
ox-LDL can induce MCP-3 expression in HUVECs. MCP-3 induces apoptosis of HUVECs and significantly affects the cellular function partially through the PI3K signaling pathway.
Department of Cardiology, General Hospital of PLA, Beijing, China. cjsong001@sina.com
Jinsong
J
Chen
Bo
B
Zhang
Congze
C
Pan
Lei
L
Ren
Yundai
Y
Chen
D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D002448:Cell Adhesion; D002478:Cells, Cultured; D054410:Chemokine CCL7 / Q000494:pharmacology*; D002867:Chromones / Q000494:pharmacology; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D008074:Lipoproteins / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000494:pharmacology; D009025:Morpholines / Q000494:pharmacology; D000081082:Phosphoinositide-3 Kinase Inhibitors; D054390:Receptors, CCR2 / Q000037:antagonists & inhibitors; D015398:Signal Transduction; D013925:Thromboplastin / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism*
D054410:Chemokine CCL7; D002867:Chromones; C573420:ICAM1 protein, human; D008074:Lipoproteins; D008077:Lipoproteins, LDL; D009025:Morpholines; D000081082:Phosphoinositide-3 Kinase Inhibitors; D054390:Receptors, CCR2; D019010:Vascular Cell Adhesion Molecule-1; C051928:lipoprotein-associated coagulation inhibitor; C099252:oxidized low density lipoprotein; D018799:Intercellular Adhesion Molecule-1; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; D013925:Thromboplastin; C505407:CASP3 protein, human; D053148:Caspase 3