Development of inhibitor for inflammatory bone resorption using neuropeptide vasoactive intestinal peptide

利用神经肽血管活性肠肽开发炎症性骨吸收抑制剂

• 批准号: 14370708
• 负责人: MUKOHYAMA Hitoshi
• 金额: $ 8.06万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370708/
• 关键词: Neuropeptide; vasoactive intestinal peptide; osteoclasts; ostebolasts; VIP receptor; vasoactive intestinal peptide; neuropeptiede; vasoactive intesitinal peptide

In the present study, we examined the effect of VIP on osteoclast formation in mouse bone marrow cultures. In this culture system, 1,25(OH)_2-vitamin D3 (D3) or parathyroid hormone (PTH) induce formation of multinucleated cells positive for tartrate resistant acid phophatase (TRAP+MNC), with calcitonin binding sites and an ability to form pits on bone slices ; three phenotypic expressions of osteoclasts. VIP reduced, in a concentration-dependent manner, osteoclast formation, the number of binding sites for radiolabelled calcitonin and the amounts of resorption pits in cultures stimulated by PTH or D3. Osteoclast formation stimulated by D3 was associated with stimulations of mRNA for calcitonin receptor, cathepsin K and TRAP. The stimulatory direct by D3 on calcitonin receptor and cathepsin K mRNA, but not that on TRAP mRNA, was down regulated by VIP. VIP increased the mRNA for the osteoblastic markers alkaline phosphatase and osteopontin. The inhibition of TRAP+MNC formation by VIP was mimicked by forskolin. The presence of VIP during last two days of a 7-day-culture period was sufficient to inhibit osteoclast formation. Not only VIP, but also PACAP-38 and secretin, but not glucagon, inhibited osteoclast formation and the number of calcitonin binding sites in D3 stimulated cultures. RT-PCR revealed the expression of VIP-1 and VIP-2 receptor subtypes with the VIP-2 receptor mRNA down regulated by D3. These findings indicate that VIP, via VIP-1 and/or VIP-2 receptors, inhibits osteoclastogenesis by a cyclic AMP-dependent mechanism, due to an inhibitory effect at the late stage of osteoclastogenesis. The present study supports the idea of a neuro-osteogenic interaction in bone metabolism.

在本研究中，我们研究了VIP对小鼠骨髓培养中破骨细胞形成的影响。在此培养体系中，1，25（OH）_2-维生素D_3（D_3）或甲状旁腺激素（PTH）诱导形成抗酒石酸酸性磷酸酶（TRAP+MNC）阳性的多核细胞，具有降钙素结合位点和在骨片上形成小凹的能力; VIP减少，在浓度依赖性的方式，破骨细胞的形成，放射性标记的降钙素的结合位点的数量和数量的吸收坑在PTH或D3刺激的文化。D3刺激的破骨细胞形成与降钙素受体、组织蛋白酶K和TRAP的mRNA的刺激有关。VIP可下调D3对降钙素受体和组织蛋白酶K mRNA的直接刺激作用，但对TRAP mRNA的刺激作用不明显。VIP增加成骨细胞标志物碱性磷酸酶和骨桥蛋白的mRNA。VIP对TRAP+MNC形成的抑制作用由forskolin模拟。在7天培养期的最后两天VIP的存在足以抑制破骨细胞的形成。不仅VIP，而且PACAP-38和胰泌素，但不是胰高血糖素，抑制破骨细胞的形成和降钙素结合位点的数量在D3刺激的文化。RT-PCR显示VIP-1和VIP-2受体亚型的表达，VIP-2受体mRNA被D3下调。这些结果表明，VIP，通过VIP-1和/或VIP-2受体，抑制破骨细胞的周期性AMP依赖性机制，由于在破骨细胞生成的晚期阶段的抑制作用。本研究支持骨代谢中神经-成骨相互作用的观点。