华西口腔医学杂志

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人白细胞介素1受体拮抗剂基因转染人胚颞颌关节软骨细胞的实验研究

李逸松,田卫东,王栋,李声伟,陈希哲   

  1. 610041 四川大学华西口腔医学院口腔颌面外科学教研室
  • 收稿日期:2003-02-25 修回日期:2003-02-25 出版日期:2003-02-20 发布日期:2003-02-20
  • 基金资助:

    本课题为国家教育部回国人员启动基金(编号363)及四川省青年基金(编号387)资助项目

An Experimental Study on Gene Transfection of Human Interleukin-1 Receptor Antagonist Gene into Chondrocytes of Temporomandibular Joint

LI Yisong,TIANWeidong,WANG Dong,et al.   

  1. Department ofOral andMaxillofacial Surgery,West China College ofStomatolo- gy,Sichuan University,Chengdu610041China
  • Received:2003-02-25 Revised:2003-02-25 Online:2003-02-20 Published:2003-02-20

摘要:

目的 研究人白细胞介素1受体拮抗剂(hIL-1ra)基因转染人胚颞颌关节软骨细胞的方法和表达。方法 分离培养人胚颞颌关节软骨细胞,将hIL-1ra基因以阳离子脂质体Lipefectine介导进行基因转染,免疫细胞化学染色和ELISA法检测转染细胞中hIL-1ra蛋白的表达。结果 使用阳离子脂质体介导hIL-1ra基因转染人胚颞颌关节软骨细胞,免疫细胞化学染色hIL-1ra蛋白呈阳性表达,瞬时转染和稳定转染时,转染细胞胞浆内和培养基上清液中均有目的基因的表达,与未经转染的对照组存在显著性差异(P<0·01)。结论 体外实验证实脂质体介导hIL-1ra基因转染方法切实可行,为今后颞颌关节骨关节病的基因治疗提供了依据。

关键词: 软骨细胞, 人白细胞介素1受体拮抗剂, 颞颌关节, 骨关节病, 基因转染

Abstract:

Objective The purpose of this study was to investigate methods and expression of the recombinant human inter- leukin-1 receptor antagonist gene transfected into chondrocytes of temporomandibular joints (TMJ).Methods Chondrocytes derived from the TMJ of 5~7 months old human fetus were transfected by recombinant human interleukin-1 receptor antagonist gene via cat- ionic liposome (Lipefectine) as a medium. The expressing level of hIL-1ra protein was detected using immunocytochemistry and enz- yme-linked immunosorbent assay (ELISA).Results The positive results of the immunocytochemistry were found. The positive ex- pression was detected in the cell plasma and the cell culture supernatant, and there was significant difference between the cells with and without gene transfection (P<0.01).Conclusion This feasible method provides experimented evidence for the future gene therapy of temporomandibular joint osteoarthrosis.

Key words: chondrocyte, human interleukin-1 receptor antagonist, temporomandibular joint, osteoarthrosis, gene transfection