中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (28): 5136-5140.doi: 10.3969/j.issn.1673-8225.2010.28.002

• 骨组织构建 bone tissue construction • 上一篇    下一篇

低氧对小鼠破骨细胞分化影响的时间依赖关系

郎红梅，金小岚，万  勇，游志清   

  1. 解放军成都军区总医院内分泌科，四川省成都市   610083
  • 出版日期:2010-07-09 发布日期:2010-07-09
  • 通讯作者: 金小岚，博士，主任医师，解放军成都军区总医院内分泌科，四川省成都市 610083 williamsjin@sina.com
  • 作者简介:郎红梅★，女， 1977年生，四川省乐至县人，汉族，2007年解放军第三军医大学毕业，硕士，医师，主要从事骨代谢方面的研究。 junialang@sina.com

Time-depended effect of low oxygen on mouse osteoclast differentiation

Lang Hong-mei, Jin Xiao-lan, Wan Yong, You Zhi-qing   

  1. Department of Endocrinology, General Hospital of Chengdu Military Command, Chengdu 610083, Sichuan Province, China
  • Online:2010-07-09 Published:2010-07-09
  • Contact: Jin Xiao-lan, Doctor, Chief physician, Department of Endocrinology, General Hospital of Chengdu Military Command, Chengdu 610083, Sichuan Province, China williamsjin@sina.com
  • About author:Lang Hong-mei★, Master, Physician, Department of Endocrinology, General Hospital of Chengdu Military Command, Chengdu 610083, Sichuan Province, China junialang@sina.com

PDF

484

被引次数

22

摘要：

背景：平原人群到高原后不久会引起骨量丢失，在高原发生骨折会引起延迟愈合甚至不愈合。与低海拔地区相比，高海拔地区骨吸收增加，骨量减少。
目的：通过观察不同时间点低氧对小鼠破骨细胞分化的影响，探讨低氧对破骨细胞分化影响可能的分子机制。
方法：取小鼠骨髓细胞，用分化培养基分别在20%氧体积分数(常氧组)、和3%氧体积分数(低氧组)中培养。骨髓细胞纯化后加入到第3~5代成骨细胞进行共培养，随机分组，每组样本数为4，培养第1，3，5，7天，用TRIzol提取细胞总RNA，用半定量反转录PCR检测骨保护素、RANK及RANKLmRNA的表达；用western blotting检测RANK蛋白的表达情况。
结果与结论：与常氧组相比，骨保护素mRNA的表达随着缺氧时间的延长而降低，RANKmRNA和RANK蛋白的表达随着缺氧时间的延长而升高，RANKLmRNA的表达随着缺氧时间的延长而升高。结果表明，低氧可促进破骨样细胞的分化，尤其在缺氧第5天，破骨细胞生成最多。

关键词: 低氧, 破骨细胞, 分化, 骨保护素, RANK, RANKL, 骨组织工程

Abstract:

BACKGROUND: When the people living on the plain go to the plateau, soon they would suffer low bone mass, if a bone fracture happens on the plateau, bone heal would be delayed, even nonunion. Comparing with the region of low altitude, bone absorption in the region of high altitude increases and bone mass decreases.
OBJECTIVE: To observe the effect of lower oxygen concentration on the differentiation of mouse osteoclast at different time points, and to study the possible molecule mechanisms of effect of hypoxia on the osteoclast differentiation.
METHODS: The mouse bone marrow cells were obtained from mice and were cultured in normoxia (20% oxygen concentration) and hypoxia (3% oxygen concentration) groups. The 3rd -5th generation mature osteoblasts were added to purified bone marrow cells. These cells were randomized into 2 groups with 4 samples in each group. At 1, 3, 5 and 7 days after cultivation, total cellular RNA was isolated using total RNA kit. RT-PCR was performed to detect the expression of osteoprotegerin and RANK and RANKLmRNA. The expression of RANK protein was assayed by Western blot.
RESULTS AND CONCLUSION: Compared with normoxia group, with time prolonged, the expression of osteoprotegerin mRNA was decreased but the expression of RANKmRNA, RANKLmRNA and RANK protein were increased. The results demonstrated that hypoxia can stimulate the differentiation of osteoclasts. Especially at the 5th day of lack of oxygen, the quantity of osteoclasts formation was greatest.

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