南方医科大学学报

南方医科大学学报 ›› 2013, Vol. 33 ›› Issue (01): 86-.

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MCP-3对人脐静脉内皮细胞表达ICAM-1、VCAM-1、TF/TFPI及其凋亡的影响

陈劲松，张波，潘从泽，任磊，陈韵岱   

  • 出版日期:2013-01-20 发布日期:2013-01-20

Effects of monocyte chemotactic protein-3 on ICAM-1, VCAM-1, TF, and TFPI expression and apoptosis in human umbilical vein endothelial cells

  • Online:2013-01-20 Published:2013-01-20

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摘要： 目的研究单核细胞趋化因子-3（MCP-3）对人脐静脉内皮细胞（HUVECs）表达ICAM-1、VCAM-1、TF、TFPI 及其凋亡
的影响。方法分离、培养人脐静脉内皮细胞，分别给予0~3.0 ng/ml的MCP-3进行干预，确定MCP-3对HUVECs的最适作用
浓度，以此浓度干预HUVECs，在MCP-3 加药之前1 h 分别给予MCP-3 抗体（20 ng/ml）、PI3K抑制剂LY-294002（5 mmol/ml）
处理细胞，MCP-3 作用24 h 后提取各组总RNA、总蛋白，用RT-PCR、Western blot 分别检测干预后的ICAM-1、VCAM-1、TF、
TFPI 的表达。模拟病理状态，用ox-LDL 干预HUVECs与空白组对照，24 h 后提取各组总RNA、总蛋白，用RT-PCR、Western
Blot 分别检测各组MCP-3 表达情况。最后，用MCP-3 及MCP-3+CCR2 拮抗剂RS102895（6μmol/L）分别干预HUVECs，作用
24、48 h 分别检测HUVECs凋亡率及caspase3 的表达，并与空白对照组比较。结果MCP-3 对HUVECs的最适作用浓度为
0.3 ng/ml（P<0.05）；MCP-3 可诱导HUVECs 细胞中ICAM-1、VCAM-1、TF 表达的增加，同时可抑制TFPI 表达（P<0.05）；
MCP-3 抗体和LY-294002 可分别拮抗、抑制此作用（P<0.05）；ox-LDL 可上调HUVECs表达MCP-3（P<0.05）；作用24、48 h 后
MCP-3 可明显诱导HUVECs凋亡，CCR2 抑制剂可拮抗此作用。结论ox-LDL 可诱导HUVECs表达MCP-3，MCP-3 可诱导
HUVECs凋亡，且可能部分通过PI3K信号通路影响HUVECs功能。

Abstract: 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. Conclusion 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.