中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (42): 7819-7824.doi: 10.3969/j.issn.2095-4344.2012.42.006

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

缺氧大鼠成骨细胞增殖、分化及基因表达

顾九君1，盛俊东2，梁伟东1   

  1. 1兰州市第一人民医院骨科，甘肃省兰州市 730050
    2天水市第一人民医院脊柱外科，甘肃省天水市 741000
  • 收稿日期:2012-02-01 修回日期:2012-03-13 出版日期:2012-10-14 发布日期:2012-10-14
  • 作者简介:顾九君★，男，1975年生，甘肃省皋兰县人，汉族，2007年兰州大学医学院毕业，硕士，主治医师，主要从事骨质疏松的基础与临床研究。 Gujiujun57343@163.com

Proliferation, differentiation and gene expression of osteoblasts in hypoxic rats

Gu Jiu-jun1, Sheng Jun-dong2, Liang Wei-dong1   

  1. 1Department of Orthopedics, First People’s Hospital of Lanzhou, Lanzhou 730050, Gansu Province, China
    2Department of Spinal Surgery, First People’s Hospital of Tianshui, Tianshui 741000, Gansu Province, China
  • Received:2012-02-01 Revised:2012-03-13 Online:2012-10-14 Published:2012-10-14
  • About author:Gu Jiu-jun★, Master, Attending physician, Department of Orthopaedics, First People’s Hospital of Lanzhou, Lanzhou 730050, Gansu Province, China Gujiujun57343@163.com

PDF

347

被引次数

7

摘要：

背景：研究表明，低氧会引起骨折延迟愈合或不愈合，骨密度减低，使骨质疏松、骨折等疾病的发病率升高。成骨细胞是骨形成、生长和发育主要的功能细胞。
目的：观察缺氧对体外培养成骨细胞增殖、分化及基因表达的影响。
方法：选用新生Wistar大鼠颅盖骨，使用胰酶-胶原酶序贯消化法获取成骨细胞，进行体外传代培养及鉴定。在缺氧培养下应用MTT法测定成骨细胞增殖率，对硝基苯磷酸盐法测定成骨细胞碱性磷酸酶活性，反转录-聚合酶链反应法测定成骨细胞内骨钙素及Ⅰ型胶原的表达。
结果与结论：缺氧具有抑制成骨细胞增殖，降低碱性磷酸酶活性及下调大鼠成骨细胞中Ⅰ型胶原α1、骨钙素基因表达的作用，随缺氧时间的增加作用更加明显。提示缺氧可通过抑制成骨细胞增殖、分化成熟及下调Ⅰ型胶原α1、骨钙素基因表达而降低成骨能力，从而促进骨质疏松的发生。

关键词: 缺氧, 成骨细胞, 增殖, 分化, 基因表达, 大鼠, 组织构建

Abstract:

BACKGROUND: Several researches have shown that hypoxia can lead to the healing of fracture delayed or non healing, and reduce bone density, which will improve the incidence of osteoporosis and fracture.
OBJECTIVE: To investigate the effects of hypoxia on the proliferation, differentiation and gene expression of osteoblasts cultured in vitro.
METHODS: The cranium from a newborn Wistar rat was collected and osteoblasts were extracted by trypsogen-collagenase sequential digestion method. The cells were subcultured in vitro and identified. The reproductive rate of osteoblasts was tested by MTT assay. Alkaline phosphatase activity of osteoblasts was detected by nitrophenylphosphate method. Bone gamma-carboxyglutamic-acid-containing proteins (BGP) and type Ⅰ collagen expression were measured by reverse transcription-PCR method.
RESULTS AND CONLUSION: Our studies revealed that hypoxia could inhibit the proliferation of osteoblasts and reduce alkaline phosphatase activity as well as decrease the expression of BGP mRNA and type Ⅰ collagen mRNA in a time-dependent manner. These findings suggest that hypoxia can inhibit the proliferation and differentiation of osteoblasts cultured in vitro and decrease the expression of BGP mRNA and type Ⅰ collagen, which will decrease osteogenic ability and promote the incidence of osteoporosis.

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