C-C Motif Chemokine 8 promotes angiogenesis in vascular endothelial cells
C-C Motif Chemokine 8 promotes angiogenesis in vascular endothelial cells
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DOI：
10.1177/1708538120959972
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发表时间：
2020-09
期刊：
影响因子：
1.1
通讯作者：
Song Xue;Hanfei Tang;Gefei Zhao;Yang Shen;E. Y. Yang;W. Fu;Zhenyu Shi;Xiao Tang;D. Guo
中科院分区：
文献类型：
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作者：
Song Xue;Hanfei Tang;Gefei Zhao;Yang Shen;E. Y. Yang;W. Fu;Zhenyu Shi;Xiao Tang;D. Guo
目的血管生成是一个重要的进展与一些病理情况。已经报道了几种趋化因子作为血管生成的调节剂。本研究旨在探讨C-C基序趋化因子8是否参与血管生成的调控。方法收集颈动脉重度狭窄患者颈动脉斑块，采用CD 31免疫组化和实时荧光定量PCR方法检测斑块中C-C基序趋化因子8（C-C Motif Chemokine 8）的表达，探讨C-C Motif Chemokine 8与斑块血管生成的关系。为了进一步阐明C-C基序趋化因子8与血管生成之间的关系，在存在或不存在C-C基序趋化因子受体2-Ab和细胞外调节的MAP激酶1/2抑制（FR 180204）的情况下，用C-C基序趋化因子8处理人脐静脉内皮细胞和人真皮微血管内皮细胞。分别采用细胞计数试剂盒-8和Transwell小室法检测人脐静脉内皮细胞和人真皮微血管内皮细胞的增殖和迁移。采用试管形成试验检测C-C基序趋化因子8刺激的体外血管生成。体外和体内血管生成分别通过小鼠主动脉环试验和基质胶塞试验进行评估。采用实时荧光定量PCR检测人脐静脉内皮细胞C-C基序趋化因子受体，采用Western印迹法检测C-C基序趋化因子受体1、C-C基序趋化因子受体2、细胞外调节MAP激酶1/2和磷酸化细胞外调节MAP激酶1/2。结果C-C Motif趋化因子8在颈动脉粥样硬化斑块中表达增加，且伴有严重的血管新生。C-C基序趋化因子8（5 ng/ml）微弱地增加人脐静脉内皮细胞增殖，但不增加人真皮微血管内皮细胞增殖。C-C基序趋化因子8可诱导人脐静脉内皮细胞和人真皮微血管内皮细胞迁移和管腔形成。在小鼠主动脉环试验和基质胶栓塞试验中，C-C基序趋化因子8与载体组相比可促进血管生成。细胞外调节的MAP激酶1/2的磷酸化随着C-C基序趋化因子8刺激而增加。C-C基序趋化因子8促进的迁移和管形成可以在很大程度上被C-C基序趋化因子受体2-Ab或细胞外调节的MAP激酶1/2抑制剂阻断（FR 180204）。结论C-C基序趋化因子8在体内外均能促进血管生成。C-C基序趋化因子受体2在C-C基序趋化因子8的激活中起重要作用，细胞外调节的MAP激酶1/2信号通路参与了这一机制。
Objectives Angiogenesis is an important progress associated with several pathological situations. Several chemokines have been reported to act as regulators of angiogenesis. The current study aimed to find whether C-C Motif Chemokine 8 is involved in angiogenesis regulation. Methods To verify whether C-C Motif Chemokine 8 is related to angiogenesis in plaques, carotid plaques were collected from patients with severe carotid stenosis and analysed using CD31 immunohistochemistry and real-time PCR. To further clarify the relation between C-C Motif Chemokine 8 and angiogenesis, human umbilical vein endothelium cells and human dermal microvascular endothelial cells were treated with C-C Motif Chemokine 8 in the presence or absence of C-C motif chemokine receptor 2-Ab and extracellular regulated MAP kinase 1/2 inhibition (FR180204). Proliferation and migration of human umbilical vein endothelium cells and human dermal microvascular endothelial cells were examined with Cell Counting Kit-8 and Transwell chamber assay, respectively. In vitro angiogenesis stimulated by C-C Motif Chemokine 8 was examined using tube formation assay. Ex vivo and in vivo angiogenesis were assessed by mice aortic ring assay and Matrigel plug assay, respectively. C-C motif chemokine receptors of human umbilical vein endothelium cells were examined with real-time PCR, and C-C motif chemokine receptor 1, C-C motif chemokine receptor 2, extracellular regulated MAP kinase 1/2 and phosphorylation-extracellular regulated MAP kinase 1/2 were examined with western blotting assay. Results C-C Motif Chemokine 8 was increased in carotid plaques with severe angiogenesis in both RNA and protein level. C-C Motif Chemokine 8 (5 ng/ml) weakly increased human umbilical vein endothelium cell proliferation, but not on human dermal microvascular endothelial cells. Migration and tube formation could be induced by C-C Motif Chemokine 8 in both human umbilical vein endothelium cells and human dermal microvascular endothelial cells. In mice aortic ring assay and Matrigel plug assay, C-C Motif Chemokine 8 could promote angiogenesis compared to vehicle groups. Phosphorylation of extracellular regulated MAP kinase 1/2 was increased with C-C Motif Chemokine 8 stimulation. The migration and tube formation promoted by C-C Motif Chemokine 8 could be largely blocked by C-C motif chemokine receptor 2-Ab or extracellular regulated MAP kinase 1/2 inhibition (FR180204). Conclusions C-C Motif Chemokine 8 could promote both in vitro and in vivo angiogenesis. C-C motif chemokine receptor 2 played an important role in the activation of C-C Motif Chemokine 8 and extracellular regulated MAP kinase 1/2 signalling pathway was involved in this mechanism.