组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2019, Vol. 15 ›› Issue (2): 77-80.doi: 10.3969/j.issn.1673-0364.2019.02.004

• 论著 • 上一篇    下一篇

JNK抑制剂对软骨细胞外基质合成的作用研究

刘延群，贾立涛，刘春燕，陈洁，周广东   

  1. 潍坊医学院整形外科研究所；济南市第四人民医院；上海交通大学医学院附属第九人民医院整复外科上海市组织工程研究重点实验室；组织工程国家工程研究中心
  • 收稿日期:2018-12-27 修回日期:2019-03-02 出版日期:2019-04-20 发布日期:2019-04-20
  • 作者简介:周广东(E-mail:guangdongzhou@126.com)。

Effect of JNK Inhibitor on the Synthesis of Extracellular Matrix of Chondrocytes

LIU Yanqun,JIA Litao,LIU Chunyan,CHEN Jie,ZHOU Guangdong   

  1. Shanghai Key Laboratory of Tissue Engineering,Shanghai Ninth People's Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200011,China;The Fourth Hospital of Ji'nan,Ji'nan 250031,China; Department of Plastic and Reconstructive Surgery)
  • Received:2018-12-27 Revised:2019-03-02 Online:2019-04-20 Published:2019-04-20
  • Contact: 国家自然科学基金项目(81871502)；国家自然科学基金项目(81701843)

摘要： 目的 探索JNK抑制剂(SP600125)对软骨细胞外基质合成的影响。方法 获取、培养兔耳软骨细胞,通过CCK-8法测定SP600125对细胞增殖的影响。将兔耳软骨细胞接种于PGA/PLA支架,构建细胞-支架复合物,并随机分为实验组与对照组。实验组将细胞-支架复合物置于含有10%FBS和SP600125的DMEM中培养,对照组在含有10%FBS的DMEM中培养。体外培养8周后,RT-PCR检测IGF、COLⅡ、COLⅨ、TGF-β1等软骨特异性基因的表达,并行HE染色与Safranin-O染色。结果 CCK-8法检测显示,相较对照组,实验组软骨细胞的增殖明显增强;体外培养8周后,实验组软骨特异性基因IGF、COLⅡ、COLⅨ、TGFβ1的表达显著下降(P<0.05);组织学观察显示,实验组软骨陷窝的成熟度低,细胞外基质分泌减少。结论 SP600125可降低软骨细胞外基质的合成。

关键词: 骨关节炎, 软骨再生, 组织工程

Abstract: Objective To explore the effect of JNK inhibitor(SP600125)on the synthesis of extracellular matrix of chondrocytes.Methods Rabbit ear chondrocytes were obtained and cultured,and the effect of SP600125 on cell proliferation was determined by CCK-8 method.Rabbit ear chondrocytes were inoculated into PGA/PLA scaffolds,and cell-scaffold complexes were constructed and randomly divided into 2 groups:experimental group and control group.In the experimental group,the cell-scaffold complex was cultured in DMEM containing 10%FBS and SP600125,and the control group was cultured in DMEM containing 10%FBS only.After 8 weeks of in vitro culture,the expression of cartilage-specific genes such as IGF,COLⅡ,COLⅣand TGF-β1 was detected by RT-PCR,and HE staining and Safranin-O staining were performed.Results Compared with control group,the proliferation of chondrocytes in the experimental group was significantly promoted detected by CCK-8.After 8 weeks of in vitro culture,the expression of cartilage-specific genes IGF,COLⅡ,COLⅣand TGF-β1 in the experimental group was significantly lower than that in the control group(P<0.05).Histological observation showed that compared with the control group,the experimental group had a low degree of maturity and a decrease in extracellular matrix secretion.Conclusion SP600125 can reduce the synthesis of extracellular matrix of cartilage.

Key words: Osteoarthritis, Cartilage regeneration, Tissue engineering

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