A role of protein phosphatase 2C on osteoclast differentiation and activation

蛋白磷酸酶2C对破骨细胞分化和活化的作用

• 批准号: 16580078
• 负责人: OHNISHI Motoko
• 金额: $ 2.3万
• 依托单位: Chubu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16580078/
• 关键词: signal transduction; development, differentiation; enzyme; biological activity; cells, tissues; 破骨細胞; プロテインホスファターゼ2C; 破骨細胞分化誘導因子; RANKL

The receptor activator of NF-κB ligand (RANKL), which is a member of the tumor necrosis factor family, plays a key role in the differentiation, activation and survival of the osteoclasts. RANKL interacts with its receptor, RANK, which is expressed on osteoclast progenitors or mature osteoclasts, and then induce activation of downstream molecules such as NF-κB and p38 MAPK. Since protein phosphatase 2C (PP2C) has been reported as one of the negative regulators in SAPK/p38 signaling pathway, we examined the possibility if PP2C might be involved in osteoclastogenesis through the regulation of RANKL/RANK signaling pathway. The following conclusions can be drawn from the results of this research project.1;When human embryonic kidney epithelial 293 cells were transfected with a full-length RANK(a receptor of RANKL) expression plasmids, phosphorylation level of p38 MAPK and NF-κB activity were upregulated. We found that coexpression of PP2C suppressed activation of both p38 MAPK and NF-κB.2;We generated 293 cells, which stably express Myc epitope-tagged RANK, called 293-RANK cells. When these cells were stimulated with RANKL, increase in phosphorylation level in p38 MAPK and activation of NF-κB were observed. Overexpression of PP2C inhibited these RANKL-induced p38 phosphorylation and NF-κB activation.3;RAW264 cells, which are macrophage/monocyte cell line, are known to differentiate into osteoclast-like multinucleated cells following RANKL stimulation. Total mRNAs were prepared from RAW264 cells at regular intervals after RANKL stimulation and quantitative realtime PCR was performed in order to analyze PP2C expression. As a result, it was shown that PP2C mRNA levels increased transiently following RANKL stimulation.Taken together, our results suggest the possibility that PP2C might be concerned with the regulation of osteoclast differentiation.

NF-κB配体（RANKL）的受体激活剂是肿瘤坏死因子家族的成员，在破骨细胞的分化，激活和存活中起关键作用。 RANKL与其受体Rank相互作用，Rank在破骨细胞祖细胞或成熟的破骨细胞上表达，然后影响下游分子（例如NF-κB和p38 MAPK）的激活。由于蛋白质磷酸酶2C（PP2C）已被报道为SAPK/p38信号通路中的负调节剂之一，因此我们检查了是否可能通过RANKL/RANK信号通路来调节PP2C参与破骨细胞构成的可能性。可以从该研究项目的结果中得出以下结论。1；当人类胚胎肾上皮293细胞用全长等级（RANKL的受体）表达质粒转染时，p38 MAPK和NF-κB活性的磷酸化水平更新。我们发现，PP2C的共表达抑制了p38 MAPK和NF-κB.2; 2; 2;我们产生了293个细胞，这些细胞稳定地表达了MYC表位标记的等级，称为293级细胞。当用RANKL刺激这些细胞时，观察到p38 MAPK中磷酸化水平的增加，并观察到NF-κB的激活。 PP2C的过表达抑制了这些RANKL诱导的p38磷酸化和NF-κB激活。3; RAW264细胞是巨噬细胞/单核细胞系，已知可以区分RANKL刺激后分化为骨细胞样的多核细胞。 RANKL刺激后，定期从RAW264细胞中制备总mRNA，并进行定量实时PCR以分析PP2C表达。结果，结果表明，PP2C mRNA水平在RANKL刺激后瞬时增加。结合在一起，我们的结果表明PP2C可能与破骨细胞分化有关。

项目成果

Inhibitory effects of mevastatin and a geranylgeranyl transferase I inhibitor (GGTI-2166) on mononuclear osteoclast formation induced by receptor activator of NF kappa B ligand (RANKL) or tumor necrosis factor-alpha (TNF-alpha).

美伐他汀和香叶基香叶基转移酶 I 抑制剂 (GGTI-2166) 对 NF kappa B 配体 (RANKL) 受体激活剂或肿瘤坏死因子-α (TNF-α) 诱导的单核破骨细胞形成的抑制作用。

• 发表时间: 2005
• 期刊: Biochemical pharmacology 69
• 作者: Fukuwatari T;Ohsaki S;Fukuoka S;Sasaki R;Shibata K;福岡 伸一;福岡 伸一;Notoya M他;Woo JT 他;Notoya;Woo JT;大西素子他;Woo JT 他;Ohnishi M;Woo JT
• 通讯作者: Woo JT

Isolation of a new antibiotic, Alaremycin, structurally related to 5-aminolevulic acid from Streptomyces sp.A012304

从链霉菌属 sp.A012304 中分离出一种新抗生素 Alaremycin，其结构与 5-氨基酮戊酸相关

• 发表时间: 2005
• 期刊: Bioscience, Biotechnology, and Biochemistry 69
• 作者: Fukuwatari T;Ohsaki S;Fukuoka S;Sasaki R;Shibata K;福岡 伸一;福岡 伸一;Notoya M他;Woo JT 他;Notoya;Woo JT;大西素子他;Woo JT 他;Ohnishi M;Woo JT;Awa Y他
• 通讯作者: Awa Y他

Nonradiioactive assay of protein Ser/Thr phosphatase 2C

蛋白质 Ser/Thr 磷酸酶 2C 的非放射性测定

• 发表时间: 2005
• 期刊: Annual report of research institute for biological function 5
• 作者: Fukuwatari T;Ohsaki S;Fukuoka S;Sasaki R;Shibata K;福岡 伸一;福岡 伸一;Notoya M他;Woo JT 他;Notoya;Woo JT;大西素子他;Woo JT 他;Ohnishi M
• 通讯作者: Ohnishi M

Generation of hydrogen peroxide primary contributes to the induction of Fe(II)-dependent apoptosis in Jurkat cells by (-)-epigallocatechin gallate.

过氧化氢的产生主要有助于 (-)-表没食子儿茶素没食子酸酯诱导 Jurkat 细胞中 Fe(II) 依赖性细胞凋亡。

• 发表时间: 2004
• 期刊: Carcinogenesis 25
• 作者: Fukuwatari T;Ohsaki S;Fukuoka S;Sasaki R;Shibata K;福岡 伸一;福岡 伸一;Notoya M他;Woo JT 他;Notoya;Woo JT;大西素子他;Woo JT 他;Ohnishi M;Woo JT;Awa Y他;Woo JT 他;Kadohara K 他;Woo JT 他;Woo JT 他;Nakagawa H 他;Woo JT;Nakagawa H;Nakagawa H 他
• 通讯作者: Nakagawa H 他

Inhibitory effects of mevastatin and a geranylgeranyl transferase l inhibitor (GGTI-2166 on mononuclear osteoclast formation induced by receptor activator of NF kappa B ligand (RANKL) or tumor necrosis factor-alpha (TNF-alpha).

美伐他汀和香叶基香叶基转移酶 l 抑制剂 (GGTI-2166) 对 NF kappa B 配体 (RANKL) 受体激活剂或肿瘤坏死因子-α (TNF-α) 诱导的单核破骨细胞形成的抑制作用。

• 发表时间: 2005
• 期刊: Biochemical pharmacology 69
• 作者: Fukuwatari T;Ohsaki S;Fukuoka S;Sasaki R;Shibata K;福岡 伸一;福岡 伸一;Notoya M他;Woo JT 他;Notoya;Woo JT;大西素子他;Woo JT 他
• 通讯作者: Woo JT 他