›› 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

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