Effect of VEGF's on osteockast formation and activation

VEGF 对骨细胞形成和激活的影响

• 批准号: 13671950
• 负责人: YAMADA Shoji
• 金额: $ 1.86万
• 依托单位: Showa university
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671950/
• 关键词: VEGF; Flt-1; CSF-1; osteoclast; resorption; culture; Flk-1; TRAP; CSF-1^<op; op>マウス; VEGF受容体1型; op; opマウス; 偽足; 細胞伸展; 可溶型受容体

Vascular endothelial growth factor (VEGF) is a key mediator of physiological angiogenesis. This cytokine binds to two cell surface receptors which are encoded by flt-1 or flk-1/KDR that also belong to the tyrosine kinase receptor family. Only the Flt-1 is expressed on macrophage monocyte lineage. In the present study, we investigated that whether the effect of VEGF treatment on the osteoclast differentiation in vitro. Bone marrow cells and spleen cells were obtained from 5-to 8-week-old male ddy miceor op/op mice. Non adherent G10-passed cells were cultured in & alpha-MEM with 15% FBS, 100 ng/ml RANKL, 20 ng/ml GSF-1 or 100 ng/ml VEGF. After culture for 7 days, smaller size of TRAP positive multinucleated cells were observed in the presence of RANKL and VEGF from bone marrow cells and spleen cells(Wt & op), as well as resorption pits were found than that of in the presence of RANKL and CSF-1. Treatment with Flt-1/Fc chimera, and anti VEGF antibody inhibit the RANKL and VEGF-induced osteoclast differentiation but neutralizing anti-CSF-1 Receptor antibody did not. In addition, Flt-1 was detected by immunocytochemical study with anti-Flt-1 antibody but not Flk-1 on osteoclast and its precursors. These results suggest that VEGF has important role in osteoclast differentiation and bone resorption system as well as CSF-1.

血管内皮生长因子（VEGF）是生理性血管生成的关键介质。该细胞因子结合两种细胞表面受体，其由也属于酪氨酸激酶受体家族的flt-1或flk-1/KDR编码。只有Flt-1在巨噬细胞单核细胞系上表达。本研究旨在探讨VEGF对体外培养的破骨细胞分化的影响。从5- 8周龄雄性ddy小鼠或op/op小鼠获得骨髓细胞和脾细胞。在含有15%FBS、100 ng/ml RANKL、20 ng/ml GSF-1或100 ng/ml VEGF的α-MEM中培养非贴壁G10传代细胞。培养7 d后，在RANKL和VEGF存在下，骨髓细胞和脾细胞（Wt & op）中TRAP阳性多核细胞的体积均小于RANKL和CSF-1存在下的TRAP阳性多核细胞，并出现吸收陷窝。用Flt-1/Fc嵌合体和抗VEGF抗体处理抑制RANKL和VEGF诱导的破骨细胞分化，但中和性抗CSF-1受体抗体不能。另外，用抗Flt-1抗体免疫细胞化学法检测破骨细胞及其前体细胞上Flt-1的表达，而未检测到Flk-1。这些结果表明，VEGF在破骨细胞分化和骨吸收系统中与CSF-1一样具有重要作用。

项目成果

Mori H, Sasaki H, Morihata H, Kawawaki J, Amano H, Yamano T, Kuno: "Regulatory mechanisms and physiological relevance of a voltage-gated H+ channel in murine osteoclasts : phorbol myristate acetate induces cell acidosis and the channel activation"J Bone M

Mori H、Sasaki H、Morihata H、Kawawaki J、Amano H、Yamano T、Kuno：“小鼠破骨细胞中电压门控 H 通道的调节机制和生理相关性：佛波醇肉豆蔻酸酯乙酸酯诱导细胞酸中毒和通道激活”J Bone

天野均, 鈴木恵子, 山田庄司: "破骨細胞におけるNa^+/Ca^<2+>交換体(NCX)の役割に関する研究"昭歯誌. 21・1. 82-85 (2001)

天野仁、铃木庆子、山田庄司：“破骨细胞中Na^+/Ca^<2+>交换器（NCX）的作用的研究”Sho Dental Journal 21・1（2001）。

Hirama Y, Morohashi T, Sano T, Maki K, Ohta A, Sakai N, Yamada S, Sasa R.: "Fructooligosaccharides prevent disorders of the femoral neck following gastrectomy in growing rats."J Bone Miner Metab.. 21・5. 294-298 (2003)

Hirama Y、Morohashi T、Sano T、Maki K、Ohta A、Sakai N、Yamada S、Sasa R.：“低聚果糖可预防生长大鼠胃切除术后股骨颈的疾病。”J Bone Miner Metab.. 21・5。 294-298 (2003)

Suzuki K, Zhu B, Rittling SR, Denhardt DT, Goldberg HA, McGulloch CA, Sodek J.: "Golocalization of intracellular osteopontin with CD44 is associated with migration, cell fusion, and resorption in osteoclasts."J Bone Miner Res.. vol.17. 1486-1497 (2002)

Suzuki K, Zhu B, Ritdling SR, Denhardt DT, Goldberg HA, McGulloch CA, Sodek J.：“细胞内骨桥蛋白与 CD44 的定位与破骨细胞中的迁移、细胞融合和吸收有关。”J Bone Miner Res.. vol

信川真智子, 山田庄司, 天野均: "破骨細胞形成に及ぼすジクロフェナックナトリウムの影響"歯科薬物療法. 20・3(印刷中).

Machiko Shinkawa、Shoji Yamada、Hitoshi Amano：“双氯芬酸钠对破骨细胞形成的影响”牙科药物治疗 20・3（出版中）。