Development of bone metabolism improvement medicines based on the analysis of signal transduction

基于信号转导分析的骨代谢改善药物的开发

• 批准号: 08557101
• 负责人: SUDA Tasuo
• 金额: $ 12.54万
• 依托单位: Showa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08557101/
• 关键词: osteoblast; osteoclast; myoblast; BMP; Smad; Estrogen; IL-7; ODF; TNFファミリー; IL-1; シグナル伝達; PHT受容体; 骨誘導因子; 筋芽細胞; NF-κB; エストロゲン; インターロイキン-7; 受容体; IL-6; gp130; 波状縁

1. Regulatory mechanism of osteoblast differentiation was studied using 02012 myoblasts treated with BMP-2. BMP-2 induced Qsteoblastic differentiation of 02012 cells through BMP receptor (BM PR) IA.Samd 1 and Smad 5 were shown to be involved in the signaling pathway of BMPR IA.2. Estrogen deficiency in mice caused by ovariectomy (OVX) results in marked bone loss and accumulation of pre-B cells in bone marrow. Treatment of mice with IL-7 which stimulates B-lymphopoiesis induced marked bone loss. IL-7 receptor knockout mice exhibited increased bone volume. These results suggest that accumulation of pre-B cells is involved in the stimulated bone resorption in OVX mice.3. Expression of mRNAs of estrogen receptor (ER) a and b was studied in various tissues in rats. EAb mRNA was highly expressed in osteoblasts. This suggests that estrogen action on bone is mediated by ERb.4. Osteoblasts express osteoclast differentiation factor (ODF) as a membrane associated factor. A cDNA encoding ODF was isolated from a cDNA library of ST2 cells. ODF was a new member of the TNF-ligand family. Osteoblasts expressed ODF in response to osteotropic factors. A soluble form of ODF (sODF) together with M-0SF stimulated osteoclast formation from spleen cells in the absence of osteoblasts.5. A new culture system for human osteoclastogenesis was established. When human peripheral blood mononuclear cells were cultured with sODF and M-CSF, human osteoclasts were formed within 7 days.6. Osteoblasts induce pit forming activity of osteoclasts placed on dentine slices. sODE also stimulated pit-forming activity of osteoclasts in the absence of osteoblasts. Activation of NF-kB by ODF appeared to be involved in the induction of osteoclast function.

1.利用BMP-2处理的02012成肌细胞研究成骨细胞分化的调控机制。 BMP-2通过BMP受体(BM PR) IA诱导02012细胞的Qsteoblastic分化。Samd 1和Smad 5被证明参与BMPR IA.2的信号通路。卵巢切除术 (OVX) 引起的小鼠雌激素缺乏会导致明显的骨质流失和骨髓中前 B 细胞的积累。用刺激 B 淋巴细胞生成的 IL-7 治疗小鼠会导致明显的骨质流失。 IL-7受体敲除小鼠表现出骨体积增加。这些结果表明前B细胞的积累参与了OVX小鼠骨吸收的刺激。3．研究了大鼠不同组织中雌激素受体 (ER) a 和 b mRNA 的表达。 EAb mRNA 在成骨细胞中高表达。这表明雌激素对骨的作用是由 ERb.4 介导的。成骨细胞表达破骨细胞分化因子（ODF）作为膜相关因子。从 ST2 细胞的 cDNA 文库中分离出编码 ODF 的 cDNA。 ODF是TNF配体家族的新成员。成骨细胞响应成骨因子表达 ODF。可溶形式的ODF (sODF) 与M-0SF 一起在没有成骨细胞的情况下刺激脾细胞形成破骨细胞。5．建立了一种新的人破骨细胞生成培养体系。人外周血单个核细胞与sODF和M-CSF联合培养，7天内即可形成人破骨细胞。 6．成骨细胞诱导放置在牙本质切片上的破骨细胞的凹坑形成活性。在没有成骨细胞的情况下，sODE 还能刺激破骨细胞的坑形成活性。 ODF 激活 NF-kB 似乎参与了破骨细胞功能的诱导。

项目成果

Suda,T.,et al.: "Pathogenesis of bone loss due to estrogen deficiency." Osteoporosis 1997. 7. S43-S46 (1997)

Suda,T.,et al.：“雌激素缺乏引起的骨质流失的发病机制。”

Suda.T.,et al.: "Vitamin D and osteoclastogenesis." Academic Press(in press), (1997)

Suda.T. 等人：“维生素 D 和破骨细胞生成”。

Onoe, Y., et al.: "Expression of estrogen receptor β in rat bone." Endocrinology. 138. 4509-4512 (1997)

Onoe, Y., et al.：“大鼠骨中雌激素受体 β 的表达。”138. 4509-4512 (1997)

Yamamoto, N., et al.: "Smad1 and Smad5 act downstream of intracellular signalings of BMP-2 that inhibits myogenic differentiation and induces osteoblast differentiation in C2C12 myoblasts." Biochem. Biophys. Res.Commun.238. 574-580 (1997)

Yamamoto, N. 等人：“Smad1 和 Smad5 在 BMP-2 细胞内信号传导下游发挥作用，抑制 C2C12 成肌细胞中的肌原分化并诱导成骨细胞分化。”

Suda,T.,et al.: "Vitamin D" Vitamin D and osteoclastogenesis., 20 (1997)

Suda,T.,et al.：“维生素 D”维生素 D 和破骨细胞生成，20 (1997)