Supperession of Osteoclast Differentiation by Protein Phosphatase 2C

蛋白磷酸酶2C对破骨细胞分化的抑制

• 批准号: 18580096
• 负责人: OHNISHI Motoko
• 金额: $ 2.46万
• 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: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18580096/
• 关键词: protein phosohatase; osteoclasts; RANKL; RANKL; プロテインホスファターゼ2C

In the former research project we established the cell line called 293-RANK cells stably expressing RANK, the receptor of RANKL, and showed that overexpressed PP2Cβ or PP2CE in 293-RANK cells suppressed the activation of p38 and NF-κB induced by RANKL. Based on those results, further investigation was performed to elucidate possible biological roles of PP2Cβ and PP2Cε in osteoclastogenesis. The following conclusions can be drawn from the results of this research project.1. Overexpression of PP2Cβ or PP2Cε suppressed activation of JNK as well p38 but not ERK, induced by RANKL. RANKL stimulated activation of MKK6 located upstream of p38 and TAKI, which is an upstream kinase of MKK6, was also suppressed by overexpression of PP2CP or PP2Cε. This indicates that PP2Cβ or PP2Cε can suppress RANKL/RANK signaling pathway via inactivation of TAM.2. RAW264 cells, which form osteoclast-like cells by treatment with RANKL, were transiently transfected with plasmids encoding PP2Cβ siRNA. With RANKL treatment, TRAP activity and number of multinucleated cells were increased in transfected cells with PP2Cβ siRNA plasmids compared to those in transfected cells with empty vectors. It is suggested that differentiation of RAW264 cells into osteoclast-like cells is promoted by inhibition of PP2Cβ.3. Because PP2Cβ knockout mice(-/-), which we generated, were lethal, we used PP2Cβ heterozygous(+/-) mice to examine if there is any difference between heterozygous(+/-) mice and wild type(+/+) littermates in osteoclastic differentiation from their bone marrow stromal cells. As a result, after treatment for 72hr with M-CSF and RANKL, TRAP activity in cells from heterozygous(+/-) mice was higher than those from their wild type(+/+) littermates. This suggests that PP2Cβ might have suppressive effect on M-CSF and RANKL mediated osteoclaast formation in vivo.These results provide strong evidence that PP2Cβ has a suppressive effect on osteoclastogenesis.

在以前的研究项目中，我们建立了称为293级细胞的细胞系，稳定表达RANKL的接收器，并表明在293级细胞中过表达的PP2Cβ或PP2CE抑制了RANKL诱导的P38和NF-κB的激活。基于这些结果，进行了进一步的研究，以阐明PP2Cβ和PP2Cε在破骨细胞发生中的可能生物学作用。可以从该研究项目的结果中得出以下结论1。 PP2Cβ或PP2Cε的过表达也抑制了JNK的激活p38，而不是由RANKL诱导的ERK。 RANKL刺激位于p38和Taki上游的MKK6激活，这是MKK6的上游激酶，也因PP2CP或PP2Cε的过表达而受到抑制。这表明PP2Cβ或PP2Cε可以通过TAM.2灭活来抑制RANKL/RANK信号通路。通过用RANKL处理形成破骨细胞样细胞的RAW264细胞用编码PP2CβIRNA的质粒瞬时翻译。通过RANKL处理，与带有空载体的翻译细胞相比，PP2CβsiRNA质粒的翻译细胞中的陷阱活性和多核细胞数量增加。建议通过抑制PP2Cβ.3促进RAW264细胞分化为破骨细胞样细胞。由于我们产生的PP2Cβ基因敲除小鼠（ - / - ）是致命的，因此我们使用PP2Cβ杂合子（+/--）小鼠检查是否有杂合（+/-）小鼠和野生型（+/+）在与骨骼骨髓细胞中的杂球分化中的杂合（+/-）小鼠（+//+）之间有任何差异。结果，在用M-CSF和RANKL治疗72小时后，来自杂合（+/-）小鼠的细胞中的陷阱活性高于其野生型（+/+）同窝仔的小鼠。这表明PP2Cβ可能对体内的M-CSF和RANKL介导的破骨剂形成具有抑制作用。这些结果提供了有力的证据，表明PP2Cβ对破骨细胞生成有抑制作用。

项目成果

Pisiferdiolのプロテインホスファターゼ2C活性化作用

Pisiferdiol 的蛋白磷酸酶 2C 激活作用

• 发表时间: 2008
• 作者: Harata M. ;et. al.;Nakagawa H 他;Yonezawa T 他;Harata M;大西 素子;原田昌彦;Motoko Ohnishi;Harata et al.;大西素子 他;Notoya M 他;Woo JT 他;藤澤 望美;吉田 真実
• 通讯作者: 吉田 真実

The role of PP2CIβ in RANKL/RANK signalling pathway.

PP2CIβ在RANKL/RANK信号通路中的作用。

• 发表时间: 2007
• 作者: Harata M. ;et. al.;Nakagawa H 他;Yonezawa T 他;Harata M;大西 素子;原田昌彦;Motoko Ohnishi;Harata et al.;大西素子 他;Notoya M 他;Woo JT 他;藤澤 望美;吉田 真実;Nozomi Fujisawa;Mami Yoshida;吉田真実;Nobuhiro Aburai;Nobuhiro Aburai;Nobuhiro Aburai;大西 素子;Motoko Ohnishi;大西素子;油井 信弘;三好 信寛;Nobuhiro Miyoshi;斉藤 圭一;油井 信弘;Nobuhiro Aburai;三好 信寛;Miyoshi Nobuhiro
• 通讯作者: Miyoshi Nobuhiro

Tributyltin and triphenyltin inhibit osteoclast differentiation through a retinoic acid receptor-dependent signaling pathway.

• DOI: 10.1016/j.bbrc.2006.12.237
• 发表时间: 2007-03
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: T. Yonezawa;S. Hasegawa;Jae-Yong Ahn;Byung‐Yoon Cha;T. Teruya;H. Hagiwara;K. Nagai;J. Woo

Suppressive effects of Anoectochilus formosanus extract on osteoclast formation in vitro and bone resorption in vivo

• DOI: 10.1007/s00774-007-0810-8
• 发表时间: 2008-03-01
• 期刊: JOURNAL OF BONE AND MINERAL METABOLISM
• 影响因子: 3.3
• 作者: Masuda, Kikuko;Ikeuchi, Mayumi;Yazawa, Kazunaga
• 通讯作者: Yazawa, Kazunaga

遺伝子変異酵母に作用するpyrrocidine類の生物活性

吡咯啶对基因突变酵母的生物活性

• 发表时间: 2008
• 作者: Harata M. ;et. al.;Nakagawa H 他;Yonezawa T 他;Harata M;大西 素子;原田昌彦;Motoko Ohnishi;Harata et al.;大西素子 他;Notoya M 他;Woo JT 他;藤澤 望美
• 通讯作者: 藤澤 望美