Role of matrix proteins in the local regulation of bone metabolism and its abonomalities in osteoporosis

基质蛋白在骨代谢局部调节中的作用及其在骨质疏松症中的异常

• 批准号: 06454336
• 负责人: MATSUMOTO Toshio
• 金额: $ 4.54万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454336/
• 关键词: matrix protein; growth factor; integrin; osteoblast; differentiation; bone formation; aging; osteoporosis; TGF-β; focal adhesion kinese; チロシンリン酸化; interleukin-11; TGF-β(transforming growth factor); アルカリフォスファターゼ

1.Regulation of osteoblastic differentiation by cell-matrix interactions : Interaction of type I collagen (COL(I)) with alpha2beta1 integrin causes differentiation and transforming growth factor (TGF) -beta receptor down-regulation in osteoblastic cells. The TGF-beta receptor down-regulation enables cells to escape from the inhibition of differentiation by TGF-beta. To clarify how the cell-matrix interaction regulates these phenotypic changes, signaling pathways were examined in murine MC3T3-E1 cells. Attachment of cells to COL (I) stimulated tyrosine phosphorylation of focal adhesion kinase (FAK) and mitogen-activated protein kinase (MAPK), and enhanced MAPK activity. Inhibition of tyrosine kinase by herbimycin A,destruction of focal adhesion by cytochalasin D,or overexpression of antisense FAK mRNA prevented the activation of MAPK and the increase in alkaline phosphatase (ALP) activity. Transient expression of a MAPK-specific phosphatase, CL100, also suppressed the elevation of ALP a … More ctivity. In contrast, TGF-beta receptor down-regulation was not affected by CL100, but was abrogated by wortmannin, a phosphatidyl inositol (PL) -3 kinase inhibitor. These results demonstrate that COL (I) -alpha2beta1 integrin interaction facilitates differentiation via FAK-MAPK pathway, and down-regulates TGF-beta receptors via FAK-PI-3 kinase pathway. These signaling pathways may play an important role in the sequential differentiation of osteoblasts during bone formation.2.Impaired interleukin (IL) -11 actions in sensence-accelerated mice (SAM) : SAM shows reduced bone formation with lower bone mineral density than its normal counterpart. Bone marrow stromal cells (BMSC) from SAM contain less osteoprogenitor cells while exhibiting enhanced adipocytic differentiation, and the formation of both osteoblasts and osteoclasts is impaired in SAM.Addition of IL-11 to BMSC from SAM suppressed adipogenesis and restored osteoblast formation. The expression of IL-11 was reduced in BMSC from SAM mice, whereas no abnormality was found in IL-11 cDNA from SAM mice. These results suggest that an insufficient expression of IL-11 in BMSC causes the impairment of osteoblast and osteoclast formation in SAM mice. Less

1.细胞-基质相互作用对成骨细胞分化的调控：I型胶原(Col(I))与α2β1整合素的相互作用导致成骨细胞分化，转化生长因子-β受体下调。转化生长因子-β受体的下调使细胞能够逃避转化生长因子-β对细胞分化的抑制。为了阐明细胞-基质相互作用如何调节这些表型变化，我们研究了小鼠MC3T3-E1细胞中的信号通路。细胞与Col(I)结合可刺激粘着斑激酶(FAK)和丝裂原活化蛋白激酶(MAPK)的酪氨酸磷酸化，增强MAPK活性。Hbimycin A抑制酪氨酸激酶，细胞松弛素D破坏局部黏附，或反义FAK mRNA过表达可阻止MAPK的激活和碱性磷酸酶(ALP)活性的增加。MAPK特异性磷酸酶CL100的瞬时表达也抑制了碱性磷酸酶a…的升高更有活力。相反，转化生长因子-β受体的下调不受CL100的影响，但可被磷脂酰肌醇(PL)-3激酶抑制剂Wortmannin取消。这些结果表明，Col(I)-α2beta1整合素相互作用通过FAK-MAPK途径促进分化，并通过FAK-PI-3激酶途径下调TGF-β受体。这些信号通路可能在成骨细胞在骨形成过程中的顺序分化中发挥重要作用。2.感觉加速小鼠(SAM)中IL-11的作用减弱：SAM表现为骨形成减少，骨密度低于正常小鼠。SAM来源的骨髓基质细胞(BMSC)含有较少的骨祖细胞，但脂肪细胞分化增强，成骨细胞和破骨细胞的形成均受到损害。在SAM来源的BMSC中加入IL-11可抑制脂肪生成并恢复成骨细胞形成。SAM小鼠BMSC中IL-11的表达降低，而SAM小鼠的IL-11基因表达未见异常。这些结果表明，IL-11在BMSC中的表达不足导致SAM小鼠成骨细胞和破骨细胞形成障碍。较少

项目成果

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H.Fuse 等人：“一种新的合成类固醇，醋酸奥斯特龙 (TZP-4238)，通过增强卵巢切除大鼠的骨形成来增加皮质骨质量和强度。”

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Toshiyuki Nakamura 等人：“通过全身注射重组碱性成纤维细胞生长因子刺激骨内骨形成”（内分泌学）。

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川口 浩・松本 俊夫: "新 図説臨床整形外科講座 第10巻 骨系統・代謝疾患" メジカルレビュー社, 323 (1994)

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H.Fuse,et al.: "A new synthesic steroid,osaterone acetate(TZP-4238),increases cortical bone mass and strength by enhancing bone formation in ovariectomized rats" J Bone Mineral Res. 12(4)(in press). (1997)

H.Fuse 等人：“一种新的合成类固醇，醋酸奥沙酮 (TZP-4238)，通过增强卵巢切除大鼠的骨形成来增加皮质骨质量和强度”J Bone Mineral Res。