人白细胞介素-1受体拮抗剂基因转染人颞颌关节软骨细胞的基因表达
Expression of Human Interleukin-1 Receptor Antagonist in the Transfected Chondrocytes of Temporomandibular Joint
作者：李逸松;田卫东;李声伟;刘流;代晓明
Author：LI Yi-song1, TIAN Wei-dong2, LI Sheng-wei2, LIU Liu1, DAI Xiao-ming1
收稿日期：2006-08-25 年卷（期）页码：2006,24(04):300-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：人白细胞介素-1受体拮抗剂,基因转染,阳离子脂质体,
Key words：human interleukin-1 receptor antagonist,gene transfection,cationic liposome,
基金项目：
国家教育部回国人员启动基金资助项目（363）；四川省
青年基金资助项目（387）
中文摘要
目的 研究人白细胞介素-1受体拮抗剂（hIL-1ra）基因转染人颞颌关节软骨细胞的基因表达情况。方法hIL-1ra基因以阳离子脂质体介导转染体外培养的人颞颌关节软骨细胞，描记细胞生长曲线，计算细胞群体倍增时间，并采用酶联免疫吸附法检测瞬时转染和稳定转染细胞中hIL-1ra蛋白的表达。结果 转染细胞与正常细胞的增殖率有一定差异，转染细胞胞浆内和培养基上清液中均有hIL-1ra蛋白的表达。在细胞转染结束后48 h，细胞内hIL-1ra蛋白的表达量最高，稳定转染细胞中基因表达最长时间达72 d以上。结论 经阳离子脂质体介导hIL-1ra基因转染人颞颌关节软骨细胞后，目的基因在细胞内有一定数量的表达。
英文摘要
Objective To investigate the expression of the human interleukin-1 receptor antagonist（hIL-1ra） in the transfected chondrocytes of temporomandibular joint（TMJ）. Methods Chondrocytes of TMJ in vitro were transfected by hIL-1ra gene via cationic liposome as a medium. The stable transfected cells were selected by G418. The proliferations of the transduced cell were examined with the growth curve, cell population doubling time. The protein expressing in different periods was detected by immunocytochemistry and enzyme-linked immunosorbent assay（ELISA）. Results The proliferation suppression of gene transfected cells fell significantly with compared to normal cells. The expression of hIL-1ra was detected in the cell plasma and the cell culture supernatant. The highest expression of IL-1ra protein was at the time of 48 hours after gene transfection. The transiently transfected cells were secreted IL-1ra protein continuously 28 days and the stably transduced cells were secreted IL-1ra protein till 72 days. Conclusion This study showed that hIL-1ra protein expressed positively in the cell plasma and the culture supernatant after gene transfection within a certain periods.
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