中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (50): 9321-9324.

• 骨组织构建 • 上一篇    下一篇

肿瘤坏死因子α诱导人髓核细胞凋亡的作用途径

董振辉，王德春   

  1. 青岛大学医学院附属医院脊柱外科，山东省青岛市  266003
  • 出版日期:2010-12-10 发布日期:2010-12-10
  • 通讯作者: 王德春，博士，教授，青岛大学医学院附属医院脊柱外科，山东省青岛市 266003 dechun-w@163.com
  • 作者简介:董振辉★，男，1982年生，河北省涿州市人，汉族，2010年青岛大学医学院毕业，硕士，主要从事脊柱外科研究。 donghui0801@gmail.com
  • 基金资助:

    国家自然基金资助项目(30672132)。课题名称：理化因素对髓核细胞凋亡的影响。

Effect of tumor necrosis factor alpha on apoptosis of human nucleus pulposus cells

Dong Zhen-hui, Wang De-chun   

  1. Department of Spinal Surgery, Affiliated Hospital of Medical College of Qingdao University, Qingdao  266003, Shandong Province, China
  • Online:2010-12-10 Published:2010-12-10
  • Contact: Wang De-chun, Doctor, Professor, Department of Spinal Surgery, Affiliated Hospital of Medical College of Oingdao University, Qingdao 266003, Shandong Province, China dechun-w@163.com
  • About author:Dong Zhen-hui★, Master, Department of Spinal Surgery, Affiliated Hospital of Medical College of Qingdao University, Qingdao 266003, Shandong Province, China Donghui0801@gmail.com
  • Supported by:

    the National Natural Science Foundation of China, No. 30672132*

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摘要：

背景：在与细胞凋亡有关的众多因素中, 肿瘤坏死因子(tumor necrosis factor，TNF)超家族成员发挥了重要的作用。但TNF是通过何种途径诱导椎间盘髓核细胞凋亡的机制尚未阐明。
目的：探讨TNF-α激活后，丝裂原活化蛋白激酶(mitogen-activated protein kinases，MAPKs)信号转导通路中P38MAPK和应激活化蛋白激酶/c-Jun氨基末端激酶(stress-activated protein kinase/c-Jun NH2-terminal kinase，JNK/SAPK)两条途径对人髓核细胞凋亡的作用。
方法：体外培养人髓核细胞，将细胞随机分成4组：TNF-α刺激组，P38MAPK阻断组，P-JNK/SAPK阻断组和对照组。采用TUNEL法检测髓核细胞凋亡情况，免疫荧光法检测P-P38MAPK和P-JNK/SAPK的表达及定位；Western Blot法检测P38MAPK，JNK/SAPK及其磷酸化形式的表达。
结果与结论：TUNEL法检测凋亡结果中，TNF-α刺激组较其他各组的凋亡细胞密度大(P < 0.01)；免疫荧光结果显示TNF-α刺激组P-P38MAPK和JNK/SAPK在细胞质和细胞核的表达高于各阻断组和对照组(P < 0.01)；Western Blot结果显示P38MAPK，P-JNK/SAPK在各组髓核细胞内均有表达，但无活化形式，TNF-α刺激组可见P-P38MAPK，P-JNK/SAPK表达，但相应阻断组无表达。结果表明，外源性TNF-α可通过P38MAPK和P-JNK/SAPK途径导致人髓核细胞凋亡。

关键词: 人髓核细胞, 信号传导, 肿瘤坏死因子α, P38丝裂原活化蛋白激酶, P-应激活化蛋白激酶/ c-Jun氨基末端激酶, 细胞凋亡

Abstract:

BACKGROUND: Tumor necrosis factor (TNF) superfamily plays an important role in apoptosis. However, the role of TNF on nucleus pulposus cells remains poorly understood.
OBJECTIVE: To investigate the effects of TNF-α on apoptosis of human nucleus pulposus cell from pathways of P38MAPK and stress-activated protein kinase/c-Jun NH2-terminal kinase (JNK/SAPK).
METHODS: The human nucleus pulposus cells were cultured in vitro and randomly divided into 4 groups: TNF-α, P38MAPK inhibition, P-JNK/SAPK inhibition and control groups. The apoptosis of nucleus pulposus cells was detected by TUNEL; the expression and location of P38MAPK and P-JNK/SAPK were determined by immumofluorescence method; and the expression of P38MAPK, JNK/SAPK, and their phosphorylations were measured by Western Blot.
RESULTS AND CONCLUSION: TUNEL results showed that there was higher density of apoptotic nucleus pulposus cells in the TNF-α group than that of the other groups (P < 0.01). Immunofluorescence showed that, compared with inhibition groups and control group, the expressions of phosphorylations of P38MAPK and JNK / SAPK were increased after treatment with TNF-α (P < 0.01). Western Blot analysis also demonstrated that P38MAPK and P-JNK / SAPK were expressed and distributed mainly in cytoplasmic and nuclear, however, there was only P-JNK / SAPK expression in the TNF–α group, but no expression could be found in the inhibition groups. TNF-α induces the apoptosis of human nucleus pulposus cell via P38MAPK and JNK / SAPK pathway.

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