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Research for role of bone morphogenetic protein in the differentiation and activation of osteoclast and osteoblast

骨形态发生蛋白在破骨细胞和成骨细胞分化和活化中的作用研究

基本信息

批准号：
07671136
负责人：
SUGIMOTO Toshitsugu
金额：
$ 1.47万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671136/
关键词：
Bone morphogenetic protein Osteoclast Osteoblast Proteoglycan Differentiation Bone remodeling Bone resorption Bone formation

项目摘要

Although the action of bone morphogenetic protein (BMP) on osteoblast differentiation has been extensively investigated, its effect on osteoclast differentiation remained unknown. In this study, in vitro effects of BMP-2 on osteoclast differentiation and bone resorption were examined. BMP-2 (1-100ng/ml) significantly stimulated bone resorption in mouse bone cell culture, whereas it did not affect the bone-resorbing activity of isolated rabbit osteoclasts. BMP-2 also stimulated osteoclast differentiation in bone cell culture. We next examined the direct effect of BMP-2 on osteoclast precursors in the absence of stromal cells, using hemopoietic blast cells derived from spleen cells. The mRNA for BMP type 1 receptor was detected in these osteoclast precursor cells. And BMP-2 stimulated osteoclast formation from these hemopoietic blast cells supported by GM-CSF.The present data were the first to indicate that BMP-2 stimulates bone resorption through both direct stimulation of osteoclast fo … More rmation and activation of mature osteoclasts, possibly via stromal cells, in vitro. Next, we present a cDNA encoding a sulfate transporter (designed as st-ob for sulfate transporter from osteoblast) that was cloned as an induced gene in osteoblasts in association with osteoblastic differentiation. Based on the fact that BMP-2 induces osteoblastic phenotypes in immature fibroblastic C3H10T1/2 cells, we performed a subtraction hybridization between BMP-2-treated and -untreated cells, and have isolated one clone induced by BMP-2 to be constantly expressed in osteoblastic cells. The deduced amino acid sequence and proposed structure of st-ob were mostly identical to human diastrophic dysplasia sulfate transporter gene product (DTDST). St-ob mRNA was abundantly expressed in the thymus, testis, calvaria and osteoblastic MC3T3-E1 cells, while its expression was faint in C3H10T1/2 cells. BMP-2 increased sulfate incorporation in C3H10T1/2 cells about two fold of the baseline level. Proteoglycan is one of major components of extracellular matrix of bone and cartilage, and they induce a large amount of sulfate. In bone, osteoblasts actively uptake sulfate to synthesize proteoglycans. The sulfate transporter system in osteoblasts is mediated partly by st-ob distinct from sat-1, and st-ob is expressed during differentiation as a fundamental feature of osteoblasts. Less

虽然骨形态发生蛋白（BMP）对成骨细胞分化的作用已被广泛研究，但其对破骨细胞分化的影响尚不清楚。在这项研究中，BMP-2对破骨细胞分化和骨吸收的体外影响进行了检查。BMP-2（1- 100 ng/ml）对小鼠骨细胞的骨吸收有明显的促进作用，而对兔破骨细胞的骨吸收活性无明显影响。BMP-2还刺激骨细胞培养中的破骨细胞分化。接下来，我们使用来自脾细胞的造血母细胞，在没有基质细胞的情况下检测BMP-2对破骨细胞前体的直接作用。在这些破骨细胞前体细胞中检测到BMP 1型受体的mRNA。BMP-2刺激这些造血母细胞在GM-CSF的支持下形成破骨细胞。目前的数据首次表明，BMP-2通过直接刺激破骨细胞， ...更多信息体外成熟破骨细胞的形成和活化，可能通过基质细胞。接下来，我们提出了一个cDNA编码硫酸盐转运蛋白（设计为st-ob的硫酸盐转运蛋白从成骨细胞），克隆成骨细胞诱导基因与成骨细胞分化。基于BMP-2诱导未成熟成纤维细胞C3 H10 T1/2成骨表型的事实，我们在BMP-2处理的和未处理的细胞之间进行了消减杂交，并分离出一个经BMP-2诱导的成骨细胞中恒定表达的克隆。st-ob的推导氨基酸序列和拟议结构与人类畸形发育不良硫酸盐转运蛋白基因产物（DTDST）基本相同。St-ob mRNA在胸腺、睾丸、颅骨和成骨细胞MC 3 T3-E1中表达丰富，而在C3 H10 T1/2细胞中表达微弱。BMP-2使C3 H10 T1/2细胞中的硫酸盐掺入增加约为基线水平的两倍。蛋白多糖是骨和软骨细胞外基质的主要成分之一，它们诱导大量的硫酸盐。在骨中，成骨细胞积极摄取硫酸盐以合成蛋白聚糖。成骨细胞中的硫酸盐转运系统部分由不同于sat-1的st-ob介导，并且st-ob作为成骨细胞的基本特征在分化期间表达。少