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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.

人类骨质疏松症的病因是多因素的，可能是由衰老、绝经后和不动等情况以及全身内分泌激素和局部生长因子和细胞因子的变化引起的。我们研究了与骨代谢相关的蛋白质的mRNA水平。通过卵巢切除和固定雌性Wistar大鼠的组合来诱导实验性骨质疏松症。进行双侧坐骨神经切除术以固定后肢。大鼠每周两次口服 1,25 (OH) 2D3 或新的合成维生素 D3 类似物 2-(3-羟基丙氧基)-1,25-二羟基维生素 D3 [ED-71]，并在第 1、2、4 和 8 周处死。 0.2mug/kg ED-71 治疗组的骨矿物质含量得以保留。 1,25 (OH)2D3 0.2mug/kg 和 ED-71 0.2mug/kg 治疗组的血清骨钙素水平在第 4 周和第 8 周时有所增加。通过Northern印迹杂交测定后肢骨骼中的骨钙素mRNA水平，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正常化，还可以通过刺激成骨细胞来防止骨质疏松性骨量减少。在使用大鼠骨肉瘤细胞（ROS 17/2.8）的体外实验中，当向培养基中添加1,25（OH）2D3或ED-71时，骨钙素mRNA水平以剂量依赖性方式增加。当细胞暴露于 26,27-F6-1,25 (OH) 2D3 [F6-D3]（1,25 (OH) 2D3 的类似物）时，细胞的骨钙素 mRNA 水平增加。当向培养基中添加 1,25 (OH) 2D3 或 F6-D3 时，细胞中的碱性磷酸酶 mRNA 和骨桥蛋白 mRNA 也会增加。总之，1,25 (OH) 2D3 和两种类似物直接刺激骨钙素、碱性磷酸酶和骨桥蛋白 mRNA 表达。