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CCL2促进大鼠原代心肌微血管内皮细胞血管形成

娄强^1,*, 刘永莲¹, 张舒曼², 李园园¹, 黄小凤¹

¹河南大学抗体药物开发技术国家地方联合实验室，河南大学基础医学院，开封 475004；²河南大学淮河临床学院，开封 475000

摘要

本文旨在探讨趋化因子CCL2在成年大鼠原代心肌微血管内皮细胞(cardiac microvascular endothelial cell, CMEC)血管形成中的作用。分离原代大鼠CMEC，用CD31和VIII因子免疫染色鉴定，观察不同时间点基质胶(Matrigel)上CMEC血管形成情况，用real-time RT-PCR和ELISA分别检测在血管形成过程中CCL2的表达和分泌情况。结果显示：(1)大鼠原代CMEC分离成功，该细胞具有形成血管能力，成网数和节点数在Matrigel处理后4 h显著增加；(2) CMEC血管形成过程中CCL2和CCR2的表达量显著上调，并且CCL2的分泌量明显增加；(3) CCL2阻断抗体和CCR2拮抗剂均可显著降低CMEC血管形成能力。上述结果提示，大鼠原代CMEC在血管形成过程中可分泌CCL2，并且CCL2-CCR2信号通路促进了CMEC血管形成。

关键词： CCL2; 大鼠原代心肌微血管内皮细胞; 血管形成

分类号：R329.28

CCL2 promotes angiogenesis of primary rat cardiac microvascular endothelial cells

LOU Qiang^1,*, LIU Yong-Lian¹, ZHANG Shu-Man², LI Yuan-Yuan¹, HUANG Xiao-Feng¹

¹National and Local Joint Laboratory for Antibody Drug Development Techniques, Basic Medical School of Henan University, Kaifeng 475004, China；²The Huaihe Clinical College of Henan University, Kaifeng 475000, China

Abstract

The aim of the present study was to investigate the role of chemokine CCL2 in angiogenesis of primary adult rat cardiac microvascular endothelial cells (CMEC). The rat CMECs were isolated and identified through morphology examination and immunostaining with CD31 and factor VIII antibodies. The angiogenesis of CMEC on Matrigel was evaluated at different time points. The expression and secretion of CCL2 during the process of angiogenesis was detected by real-time RT-PCR and ELISA, respectively. The results showed that, the primary rat CMEC was isolated successfully, and the angiogenesis of CMEC was significantly induced after Matrigel treatment for 4 h. The expression of CCL2 and CCR2 were increased during angiogenesis, and the secretion of CCL2 was detected after 2 h of angiogenesis and reached the peak concentration of 1 588.1 pg/mL after 4 h. Either CCL2 blocking antibody or CCR2 antagonist significantly reduced the angiogenesis of CMEC. These results suggest that CCL2 is secreted during the process of angiogenesis of CMEC, and CCL2/CCR2 signaling pathway may play an important role in promoting angiogenesis.

Key words: CCL2; primary rat cardiac microvascular endothelial cells; angiogenesis

收稿日期：2019-08-07　　录用日期：2020-03-05

通讯作者：娄强　　E-mail: qiang_lou@126.com

DOI: 10.13294/j.aps.2020.0050

引用本文：

娄强, 刘永莲, 张舒曼, 李园园, 黄小凤. CCL2促进大鼠原代心肌微血管内皮细胞血管形成[J]. 生理学报 2020; 72 (4): 441-448.

LOU Qiang, LIU Yong-Lian, ZHANG Shu-Man, LI Yuan-Yuan, HUANG Xiao-Feng. CCL2 promotes angiogenesis of primary rat cardiac microvascular endothelial cells. Acta Physiol Sin 2020; 72 (4): 441-448 (in Chinese with English abstract).