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缺氧对体外培养髓核细胞骨钙素水平的影响
吴源旻, 徐源俊, 王善金, 谭军
同济大学
摘要:
研究了缺氧对体外培养髓核细胞骨钙素表达的影响及意义.将人髓核细胞株进行体外培养,在显微镜下观察髓核细胞的生长形态;在传代培养 24、48、72 h时分别收集2组细胞,采用免疫组织化学SABC法检测骨钙素的表达,应用图像分析系统进行了半定量检测,光密度值显示:体外培养髓核细胞在 48、72 h时,缺氧组髓核细胞OCN表达水平均明显高于常氧组(平均光密度值分别为180.50±8.76 vs 169.29±3.04、190.87±7.06 vs 166.33±4.44,均P<0.01),且缺氧组髓核细胞骨钙素表达水平随缺氧时间延长而逐渐升高.常氧组各时间段之间髓核细胞骨钙素表达水平无统计学差异.缺氧可诱导髓核细胞骨钙素的表达上调,缺氧环境下髓核细胞的骨钙素过表达,缺氧可能对椎间盘软骨终板矿化起到促进作用.
关键词:  髓核细胞  骨钙素  缺氧  免疫组化
DOI:
分类号:
基金项目:同济大学青年优秀人才培养行动计划(2013KJ075);国家自然科学基金青年基金(81201418);浦东新区卫生系统重点学科建设
Study on the effeets of hypoxia on the level of OCN in nucleus pulposus cells
WU Yuanmin, XU Yuanjun, WANG Shanjin, TAN Jun
Dongfang Hospital,Tongji University
Abstract:
In order to studing the effects of hypoxia on the level of OCN in nucleus pulposus cells.Human nucleus pulposus cell were line cultured in vitro.The growth morphological of nucleus pulposus cells were observed under a microscope.Two groups of cells were collected respectively from 24,48,72 h in culture.Immunohistochemical SABC method was used in the OCN expression.Then IPP image analysis system was used in the semi quantitative detection results of which were expressed by optical density value.Results suggested that expression of hypoxia group of nucleus pulposus cells OCN levels were significantly higher than the normal oxygen group at 48,72 h in vitro.The average optical density values were 180.50+8.76 vs 169.29+3.04,190.87+7.06 vs 166.33+4.44,P<0.01.Hypoxia group of nucleus pulposus cells OCN level increased steadily with hypoxia time(P<0.01).There is no significant difference on between different time groups the expression level of OCN in normal oxygen nucleus pulposus cells OCN between each time group.Hypoxia can induce nucleus pulposus cells overexpress OCN.Hypoxia may contribute to intervertebral disc endplate cartilage mineralization.
Key words:  nucleus pulposus cells  hypoxia  OCN  immunohistochemistry