上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (9): 1084-.doi: 10.3969/j.issn.1674-8115.2010.09.017

• 论著（基础研究） • 上一篇    下一篇

IL-1β和TNF-α对软骨细胞基质降解的影响及相关机制研究

黄金刚1, 童海骏1, 刘宏强2, 张晓玲1,3   

  1. 1.上海交通大学 医学院/中国科学院 上海生命科学研究院健康科学研究所骨科细胞与分子生物学实验室, 上海 200025；2.山西大学 体育学院, 太原 030006；3.上海交通大学 医学院附属第九人民医院骨科 |上海市骨科内植物重点实验室, 上海 200011
  • 出版日期:2010-09-25 发布日期:2010-09-27
  • 通讯作者: 张晓玲, 电子信箱: xlzhang@sibs.ac.cn。
  • 作者简介:黄金刚（1984—）, 男, 硕士；电子信箱: jgh_87@126.com。
  • 基金资助:

    国家自然科学基金（30811120440）；上海市科委基金（08410701800，08411950400）；上海市教委重点学科建设基金（J50206）

Effects of IL-1 beta and TNF-alpha on degradation of extracellular matrix of articular chondrocytes and related mechanism

HUANG Jin-gang1, TONG Hai-jun1, LIU Hong-qiang2, ZHANG Xiao-ling1,3   

  1. 1.Laboratory of Orthopedical Cellular and Molecular Biology, Institute of Health Sciences, Shanghai Jiaotong University School of Medicine &|Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China；2.College of Physical Education, Shanxi University, Taiyuan 030006, China；3.Shanghai Key Laboratory of Orthopaedic Implant, Department of Orthopaedics, The Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
  • Online:2010-09-25 Published:2010-09-27
  • Supported by:

    National Natural Science Foundation of China, 30811120440；Shanghai Science and Technology Committee Foundation, 08410701800, 08411950400；Shanghai Education Committee Foundation, J50206

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摘要：

目的 研究白介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）对关节软骨细胞外基质降解的作用及相关分子机制。方法 体外培养大鼠原代关节软骨细胞，单独或联合使用IL-1β和（或）TNF-α刺激软骨细胞，分别设为IL-1β刺激组、TNF-α刺激组、IL-1β和TNF-α联合刺激组；另设无任何刺激的对照组。在细胞培养24、48、72 h时点，采用倒置显微镜观察软骨细胞外基质的变化；采用Real-Time PCR法检测基质金属蛋白酶-13（MMP-13）、蛋白聚糖降解酶-1（Adamts-4）和蛋白聚糖降解酶-2(Adamts-5) mRNA的表达。结果 细胞在培养48、72 h时点，显微镜观察显示对照组与TNF-α刺激组的软骨细胞外基质降解情况无明显差异；IL-1β刺激组、IL-1β和TNF-α联合刺激组细胞外基质失染、降解，细胞形态缩小，细胞间隙增宽。与对照组比较，细胞培养24 h时点，IL-1β刺激组MMP-13、Adamts-4和Adamts-5 mRNA的表达显著上调（P<0.01）；而在培养48、72 h时点有所下调。IL-1β和TNF-α联合刺激组与IL-1β刺激组比较，在培养各时点，MMP-13、Adamts-4和Adamts-5 mRNA表达差异均无统计学意义（P>0.05）。结论 IL-1β诱导软骨细胞产生大量的MMP-13、Adamts-4和Adamts-5而直接降解软骨细胞外基质；TNF-α可能不直接引起软骨细胞外基质的降解。

关键词: 骨关节炎, 基质金属蛋白酶, 蛋白聚糖酶, 白介素-1β, 肿瘤坏死因子-α

Abstract:

Objective To investigate the effects of interleukin-1β (IL-1β) and tumour necrosis factor-α (TNF-α) on the degradation of extracellular matrix of articular chondrocytes, and explore the related molecular mechanism. Methods Primary articular chondrocytes were isolated from rat articular chondrocytes, and were stimulated by IL-1β and TNF-α alone or in combination. IL-1β stimulation group, TNF-α stimulation group and IL-1β and TNF-α stimulation group were established, and control group without any stimulation was also established. After treatment for 24 h, 48 h and 72 h, the changes of extracellular matrix of articular chondrocytes were observed by inverted microscope, and Real-time PCR was employed to detect the expression of metalloproteinase-13 (MMP-13), Aggrecanases-1 (Adamts-4) and Aggrecanases-2 (Adamts-5) mRNA. Results After treatment for 48 h and 72 h, it was observed by microscope that there was no significant difference in degradation of extracellular matrix of articular chondrocytes between control group and TNF-α stimulation group, while extracellular matrix of articular chondrocytes in IL-1β stimulation group and IL-1β and TNF-α stimulation group was unstained and degraded with expanded cell spaces. Compared with control group, the expression of MMP-13, Adamts-4 and Adamts-5 mRNA in IL-1β stimulation group significantly increased after treatment for 24 h （P<0.01）, and decreased after treatment for 48 h and 72 h. There was no significant difference in the expression of MMP-13, Adamts-4 and Adamts-5 mRNA between IL-1β stimulation group and IL-1β and TNF-α stimulation group after treatment for 24 h, 48 h and 72 h （P>0.05）. Conclusion IL-1β can directly degrade extracellular matrix of articular chondrocytes through up-regulation of expression of MMP-13, Adamts-4 and Adamts-5, while TNF-α can not directly degrade extracellular matrix of articular chondrocytes.

Key words: osteoarthritis, metalloproteinase, aggrecanase, interleukin-1β, tumour necrosis factor-α