Myocardial Slit2/Robo4 expression and impact of exogenous Slit2 on proliferation and migration of cardiac microvascular endothelial cells
10.3760/cma.j.issn.0253-3758.2013.12.011
- VernacularTitle:Slit2/Robo4信号通路对小鼠心肌微血管内皮细胞增殖和迁移的影响
- Author:
Gui-Xiu CHEN
1
;
Hao-Yu WANG
;
Tao LIU
;
Ming-Tao YANG
;
Zhen-Yu ZHOU
;
Gang FENG
Author Information
1. 637000,川北医学院第二临床学院南充市中心医院心内科
- Keywords:
Myocardium;
Endothelial cells;
Cell movement;
Cell proliferation
- From:
Chinese Journal of Cardiology
2013;41(12):1034-1039
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect expression of Slit2 and Robo4 in mouse ventricular muscle blood vessel and explore the impact of exogenous Slit2 on proliferation and migrate of mouse cardiac microvascular endothelial cells.Methods Slit2 and Robo4 expression in mouse ventricular muscle blood vessel was detected by immunohistochemistry.Slit2 and Robo4 expression in cardiac microvascular endothelial cells isolated from mouse ventricular muscle were detected by euzymelinked immunosorbent assay and immunofluorescence,respectively.The effects of various concentrations exogenous Slit2 on proliferation of mouse cardiac microvascular endothelial cells was examined by CCK-8 cell proliferation kit.Transwell chamber was used to detect migration of mouse cardiac microvascular endothelial cells treated with 800 μl M199 culture medium containing 20％ FBS (negative control),10 ng/ml VEGF (positive control),100 ng/ml Slit2(Slit2) and 100 ng/ml Slit2 + 10 ng/ml VEGF (Slit2 + VEGF) and incubated for 18 h at 37 ℃ and 5％ CO2.Results Both Slit2 and Robo4 protein expressions were detected in ventricular muscle blood vessel.Slit2 protein expression was detected in mouse microvascular endothelial cells.Protein and mRNA Robo4 expressions were also evidenced in mouse microvascular endothelial cells.Proliferation of mouse cardiac microvascular endothelial cells was not affected by exogenous Slit2.Migration of mouse cardiac microvascular endothelial cells was not affected by exogenous Slit2 (22.1 ± 2.8 vs.23.2 ± 3.8 in negative control,P ＞ 0.05) and significantly enhanced by VEGF (65.3 ± 3.8,P ＜ O.05 vs.Slip2 and negative control),this effect could be blocked by cotreatment with Slip2 (29.2 ± 3.4 in Slip2 + VEGF,P ＜0.05 vs.VEGF).Conclusion Slit2 and Robo4 are expressed in mouse ventricular muscle blood vessels and cardiac microvascular endothelial cells.Exogenous Slit2 has no impact on the proliferation of mouse cardiac microvascular endothelial cells but could inhibit VEGF-induced mouse cardiac microvascular endothelial cell migration.
