mRNA levels of proteins related to bone metabolism in the experimental osteoporosis of rats.

大鼠实验性骨质疏松症中与骨代谢相关的蛋白质 mRNA 水平。

基本信息

批准号：
07671597
负责人：
OKUMURA Hideo
金额：
$ 1.28万
依托单位：
Ehime 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-07671597/
关键词：
osteoporosis rat Northern blot hybridization mRNA protein related in bone metabolism osteocalcin 実験ビタミンD_3

项目摘要

Human osteoporosis is multifactorial in its etiology, and can result from such conditions as senility, post-menopause and immobilization, as well as changes in endoclinological hormones at general and growth factors and cytokines at local sites. We investigated the mRNA levels of proteins related in bone metabolism.Experimental osteoporosis was induced by a combination of ovariectomy and immobilization in female Wistar rats. Bilateral sciatic neurectomy was performed in order to immobilize the hind limbs. Rats were orally given 1,25 (OH) 2D3 or a new synthetic vitamin D3 analogue, 2- (3-hydroxypropoxy) -1,25-dihydroxyvitamin D3 [ED-71] twice a week and sacrificed at 1,2,4 and 8 weeks. Bone mineral content was preserved in the ED-71 0.2mug/kg treated group. Serum osteocalcin levels increased in the 1,25 (OH) 2D3 0.2mug/kg and ED-71 0.2mug/kg treated groups at 4 and 8 weeks. Osteocalcin mRNA levels in the bones of the hind limbs which was determined by Northern blot hybridization and increased consistently in the ED-71 0.2mug/kg treated group. These findings suggest that ED-71 may prevent the osteoporotic decrease of bone mass not only by normalizing bone resorption but also by stimulating osteoblasts.In the in vitro experiment using rat osteosarcoma cells (ROS 17/2.8), osteocalcin mRNA levels increased in a dose-dependent manner when either 1,25 (OH) 2D3 or ED-71 was added to the medium. Osteocalcin mRNA levels of the cells increased when they were exposed to 26,27-F6-1,25 (OH) 2D3 [F6-D3], (an analogue of 1,25 (OH) 2D3). Alkaline-phosphatase mRNA and osteopontin mRNA in the cells also increased when either 1,25 (OH) 2D3 or F6-D3 was added to the medium. In summary, 1,25 (OH) 2D3 and two analogues directly stimulated osteocalcin, alkaline-phosphatase and osteopontin mRNA expression.

人骨质疏松症在其病因学上是多因素，可能是由于衰老，少年后和固定化等条件以及一般性和生长因子和局部部位的细胞因子的内科细胞激素的变化而产生的。我们研究了骨代谢中相关的蛋白质的mRNA水平。经验性骨质疏松症是通过卵巢切除术和雌性Wistar大鼠固定化的结合诱导的。进行双侧坐骨神经切除术，以固定后肢。口服1,25（OH）2D3或新的合成维生素D3模拟，2-（3-羟基丙氧基）-1,25-二羟基维生素D3 [ED-71]每周两次，每周两次，以1,2,4和8周的牺牲。骨矿物质含量保存在ED-71 0.2MUG/kg治疗组中。在4和8周的1,25（OH）2d3 0.2mug/kg和ED-71 0.2mug/kg治疗组中，血清骨钙素水平升高。在后肢的骨骼中，骨钙素mRNA水平是由北印迹杂交确定的，并且在ED-71 0.2mug/kg治疗组中持续增加。这些发现表明，ED-71不仅可以通过使骨吸收归一化，而且还可以通过刺激骨细胞刺激骨质质量减少骨质量。在使用大鼠骨肉瘤细胞（ROS 17/2.8）的体外实验中，骨钙素mRNA在剂量依赖性的方式中均与1,1,25（OH-oh-oh）添加时，骨质骨肉瘤细胞（ROS 17/2.8）。当细胞暴露于26,27-F6-1,25（OH）2d3 [F6-D3]（1,25（OH）2d3）时，细胞的骨钙mRNA水平有所增加。当将1,25（OH）2d3或F6-D3添加到培养基中时，细胞中碱性磷酸酶mRNA和骨桥蛋白mRNA也会增加。总而言之，1,25（OH）2d3和两个类似物直接刺激骨钙素，碱性磷酸酶和骨桥蛋白mRNA表达。