中国组织工程研究

中国组织工程研究 ›› 2017, Vol. 21 ›› Issue (4): 520-526.doi: 10.3969/j.issn.2095-4344.2017.04.005

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

生长因子对椎间盘终板软骨细胞抗氧化能力的影响

黄晓东1，邓国英2，王伟恒1，徐立璋1，马  俊1，叶晓健1   

  1. 1解放军第二军医大学附属长征医院脊柱外科，上海市  200433；2上海交通大学附属上海市第一人民医院，上海市  200000
  • 收稿日期:2016-11-23 出版日期:2017-02-08 发布日期:2017-03-13
  • 通讯作者: 通讯作者：叶晓健，博士，主任医师，解放军第二军医大学附属长征医院脊柱外科，上海市 200433
  • 作者简介:黄晓东，男，1991年生，福建省莆田市人，解放军第二军医大学在读博士，主要从事脊柱外科方面的研究。
  • 基金资助:

    国家自然科学基金资助项目(81472071)；国家高技术研究发展计划(863)资助项目(2013AA032203)

Effects of different growth factors on the antioxidant capacity of endplate chondrocytes

Huang Xiao-dong1, Deng Guo-ying2, Wang Wei-heng1, Xu Li-zhang1, Ma Jun1, Ye Xiao-jian1   

  1. 1Department of Spine Surgery, Shanghai Changzheng Hospital of the Second Military Medical University, Shanghai 200433, China; 2Shanghai General Hospital, Shanghai Jiao Tong University, Shanghai 200000, China
  • Received:2016-11-23 Online:2017-02-08 Published:2017-03-13
  • Contact: Corresponding author: Ye Xiao-jian, M.D., Chief physician, Department of Spine Surgery, Shanghai Changzheng of the Second Military Medical University, Shanghai 200433, China
  • About author:Huang Xiao-dong, Studying for doctorate, Department of Spine Surgery, Shanghai Changzheng Hospital of the Second Military Medical University, Shanghai 200433, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81472071; the National High-Tech Research & Development Program of China (863 Program), No. 2013AA032203

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摘要：

文章快速阅读：

文题释义：
椎板软骨：椎体终板是椎体在生长发育过程中，椎体上下面的骨骺板骨化停止后形成骨板，呈轻度凹陷，即为骨性终板。椎体终板的中央仍为一薄层透明软骨覆盖，并终生存在，即为软骨终板，上下软骨终板与髓核和纤维环连接共同构成椎间盘。主要作用是防止椎间盘髓核组织嵌入椎体，同时具有平衡分散应力的作用。
抗氧化：是指抗氧化自由基的简称。人体因为与外界的持续接触，包括呼吸(氧化反应)、外界污染、放射线照射等因素不断的在人体体内产生自由基。研究证明，人体的抗氧化系统是一个可与免疫系统相比拟的、具有完善和复杂功能的系统，机体抗氧化的能力越强，就越健康，生命也越长。

摘要
背景：维生素E的抗氧化能力众所周知，但其他生长因子是否对终板软骨细胞具有抗氧化能力，国内外未见报道。
目的：观察不同生长因子对椎间盘终板软骨细胞抗氧化能力的影响。
方法：将原代培养获取的终板软骨细胞后分为4组，即空白对照组、血清剥夺组、过氧化氢刺激组、过氧化氢刺激后+不同生长因子干预组。第4组再细分为胰岛素生长因子1组、碱性成纤维细胞生长因子组、转化生长因子β组、Forskolin组、维生素E组，共5个亚组，通过RT-PCR检测各组半胱氨酸蛋白酶3、基质金属蛋白酶13、基质金属蛋白酶3、基质金属蛋白酶抑制剂1及凝血酶4、凝血酶5等基因表达情况；通过凋亡试剂盒及流式细胞仪分析不同组内细胞凋亡情况；Western blot检测细胞的合成与分泌；使用试剂盒检测总抗氧化能力和过氧化氢含量，并进行统计学分析。
结果与结论：①不同生长因子对终板软骨细胞凋亡、分泌和抗氧化能力的影响存在差异；②总体来讲，转化生长因子β和Forskolin对过氧化氢刺激后的细胞具有进一步损伤作用，而胰岛素生长因子1和维生素E对损伤后的细胞存在保护作用。

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程
ORCID: 0000-0003-4020-5898(叶晓健)

关键词: 组织构建, 软骨细胞, 胰岛素生长因子1, 碱性成纤维细胞生长因子, 转化生长因子, Forskolin, 维生素E, 椎间盘, 终板软骨细胞, 总抗氧化能力, 过氧化氢, 国家自然科学基金

Abstract:

Abstract 
BACKGROUND: It is well-known that vitamin E holds antioxidant capacity, but whether other growth factors have the same effect on endplate chondrocytes has not yet been reported.
OBJECTIVE: To observe the effect of different growth factors on the antioxidant ability of endplate chondrocytes in the intervertebral disc.
METHODS: Endplate chondrocytes were primary cultured, and then divided into four groups, including blank control, serum deprivation, hydrogen peroxide stimulation and hydrogen peroxide stimulation combined with different growth factors groups. The 4th group was subdivided into insulin-like growth factor-1, basic fibroblast growth factor, transforming growth factor β, forskolin and vitamin E groups. The expression levels of caspase-3, matrix metalloproteinase 13 and 3, inhibitor of metalloproteinase 1 as well as thrombin 4 and 5 were detected by real-time PCR. Cell apoptosis was analyzed through apoptosis kit and flow cytometry. Cell synthesis and secretion were detected by western blot assay. The total antioxidant capacity and the hydrogen peroxide content were determined by kit, and then statistically analyzed.
RESULTS AND CONCLUSION: Different growth factors had significant differences in the endplate chondrocyte apoptosis, secretion and antioxidant capacity. To conclude, transforming growth factor β and forskolin do further damage to the cells stimulated by hydrogen peroxide, while insulin-like growth factor-1 and vitamin E expose protective effect on the injured cells.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Intervertebral Disc, Insulin-Like Growth Factor I, Transforming Growth Factor beta, Vitamin E, Tissue Engineering

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