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promotes proliferation and migration of human microvascular endothelial cells

促进人微血管内皮细胞的增殖和迁移

基本信息

批准号：
18592064
负责人：
KOBAYASHI Michiyo
金额：
$ 1.86万
依托单位：
Health Sciences University of Hokkaido
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18592064/
关键词：
Osteoprotegerin Integrin αvβ Endothelial cells 細胞死

项目摘要

Objective: Osteoprotegerin (OPG) is a key regulator of osteoclastogenesis. We recently reported that endothelial cells produce high levels of OPG in response to Porphyromonas gingivalis. We also indicated that OPG treatment protects human microvascular endothelial cells (HMVECs) from detachment and apoptotic cell death induced by gingipain-active bacterial cell extracts. These results suggest that osteoprotegerin acts as a survival factor for endothelial cells during periodontitis. However, the role of OPG in the endothelium remains unknown. In this study, we investigated the effects of OPG on HMVECs. Methods: HMVECs were incubated with or without OPG (1 u g/ml final concentration)under a serum-free condition for 4 days. Cell survival was assessed by a tetrazolium (WST-8) reduction assay. Cell migration was assessed by a wound-healing assay. The expression of integrin on the cell surface was determined using the integrin-mediated cell adhesion kit, which consists of plates coated with an anti-αvβ3 antibody. Results: Within 4 days of serum deprivation, 36.7% of OPG-treated HMVECs were viable, whereas only 20.0% of OPG-untreated HMVECs were viable (P < 0.05). Wound-healing assay revealed that within 22 h, the OPG-treated HMVECs migrated to fill approximately three-quarters of the wounded area when compared with the OPG-untreated cells that showed very little migration. Furthermore, the OPG treatment for 24 h under serum deprivation condition resulted in a significant increase in the surface expression of αvβ3 integrin on the HMVECs (P < 0.05). Conclusions: OPG protects HMVECs against serum-deprivation-induced cell death. Furthermore, OPG treatment of HMVECs promotes cell migration, and up-regulates a v N 3 expression. The αvβ3 mediates the endothelial cell survival pathway. Therefore, our results also suggest that OPG may regulate the endothelial cell survival by affecting the expression of α v β 3.

目的：护骨素(OPG)是破骨细胞生成的关键调节因子。我们最近报道，内皮细胞产生高水平的OPG，以响应牙龈卟啉单胞菌。我们还指出，OPG处理可以保护人微血管内皮细胞(HMVECs)免受牙周疼痛活性细菌细胞提取物所致的脱落和细胞凋亡的影响。这些结果表明，在牙周炎过程中，护骨素是内皮细胞的生存因子。然而，OPG在内皮细胞中的作用仍不清楚。在本研究中，我们研究了OPG对HMVECs的影响。方法：人脐静脉内皮细胞在无血清条件下加入或不加入OPG(终浓度为1ug/mlOPG)培养4d。四唑盐(WST-8)还原实验检测细胞存活率。通过伤口愈合试验评估细胞迁移情况。用整合素介导的细胞黏附试剂盒检测细胞表面整合素的表达，该试剂盒由包被抗αvβ3抗体的平板组成。结果：去血清4d内，OPG处理的HMVECs存活率为36.7%，而未处理的HMVECs存活率仅为20.0%(P&lt；0.05)。伤口愈合实验显示，在22小时内，经OPG处理的HMVECs迁移至约四分之三的创面，而未经OPG处理的HMVECs几乎没有迁移。在去血清条件下，OPG作用2 4h后，HMVEC表面αvβ3整合素的表达显著增加(P<0.0 5)。结论：OPG可保护HMVECs免受血清剥夺诱导的细胞死亡。此外，OPG处理HMVECs可促进细胞迁移，并上调vN3的表达.α-v-β-3介导内皮细胞存活途径。因此，我们的结果也提示，OPG可能通过影响α-v-β-3的表达来调节内皮细胞的存活。