Effect of NaィイD1+ィエD1/CaィイD12+ィエD1 exchanger inhibitor on osteoclast formation and activation

NaiD1+D1/CaD12+D1交换抑制剂对破骨细胞形成和活化的影响

• 批准号: 10671749
• 负责人: AMANO Hitoshi
• 金额: $ 0.32万
• 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: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10671749/
• 关键词: CSF-1; osteoclast; NCX1; PLCγ; KB-R7943; osteoporosis; intracellular CaィイD12+ィエD1; bone resorption; 骨粗鬆症; アクチンリング; 分化

NaィイD1+ィエD1/CaィイD12+ィエD1 exchanger (NCX) catalyzes the electrogenic exchange of 3 NaィイD1+ィエD1 for 1 CaィイD12+ィエD1 across the plasma membrane in many mammalian cells. We have previously reported that the NaィイD1+ィエD1/CaィイD12+ィエD1 exchange activity had an important role for CSF-1-induced osteoclast differentiation. In the present study, we sought to confirm the role of NCX1, one of the isoforms of NCX and imports CaィイD12+ィエD1 into the cells from extracellular in the reverse mode, in osteoclast differentiation.Osteoclast formation was studied in mouse bone marrow cells cultured for 6-day in the presence of sRANKLigand and CSF-1, Treatment with KB-R7943, a new NCX1 inhibitor, not only blocked the reverse mode of NCX1 activity but also the osteoclast differentiation and bone resorption induced by CSF-1. It also dose-dependently inhibited NaィイD1+ィエD1-dependent ィイD145ィエD1Ca uptake In mature osteoclasts. However, Ouabain, a NaィイD1+ィエD1KィイD1+ィエD1ATPase inhibitor stimulated osteoclast formation.In addition, NCX1 antisense oligodeoxynucleotide (ODN) reduced the number of tartrate-resistant acid phosphatase-positive multinucleated cells and decreased the amount of NCX1, while non-sense or mismatched ODN did not.In conclusion, the activation of NCX1 is essential for CSF-1-induced osteoclast differentiation.

在许多哺乳动物细胞中，NaィイD 1+ィエD 1/CaィイD12+ィエD 1交换器(NCX)催化3 NaィイD 1+ィエD 1跨膜电生换为1 CaィイD12+ィエD 1。我们先前已经报道，NaィイD 1+ィエD 1/CaィイD12+ィエD 1交换活性在脑脊液-1诱导破骨细胞分化过程中起重要作用。在本研究中，我们试图证实NCX1的一种亚型NCX1在破骨细胞分化中的作用，并从细胞外反向导入CaィイD12+ィエD1。在sRANKLigand和csf-1存在下，培养6天的小鼠骨髓细胞研究了破骨细胞的形成，新的ncx1抑制剂KB-R7943不仅阻断了ncx1活性的逆转模式，而且还阻断了csf-1诱导的破骨细胞分化和骨吸收。它还剂量依赖性地抑制成熟破骨细胞对NaィイD 1+ィエD 1依赖的ィイD145ィエD1Ca的摄取。而哇巴因，NaィイD1+ィエD1KィイD1+ィエD1ATPase抑制剂可刺激破骨细胞的形成。此外，NCX1反义寡核苷酸减少了酒石酸抗性酸性磷酸酶阳性多核细胞的数量，减少了NCX1的数量，而非正义或错配的ODN则没有。

项目成果

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Hisamitsu T.、Ohata H.、Kawanishi T.、Iwamoto T.、Shigekawa M.、Amano H.、Yamada S. 和 Momose K.：“NaiID1+IeD1/CaiiD12+Ie”D1 交换器与 CaiD12+D1 的功能耦合从豚鼠回肠培养细胞的细胞内储存中释放。

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