摘要
目的研究慢病毒介导生存素基因(Lentivirus-Survivin)对早、晚期反分化人椎间盘髓核细胞的生物学效应。方法应用组织块法分离培养人椎间盘髓核细胞,采用绿色荧光素标记法检测慢病毒载体对其转染效率。通过免疫荧光法、MTT法、Western-Blot法和Antonopulos法检测比较Lentivirus-Survivin对反分化早期和晚期椎间盘髓核细胞Survivin表达、细胞活性、II型胶原和蛋白多糖合成的影响。结果反分化早期髓核细胞中存在Survivin的表达,而反分化晚期髓核细胞中则极少表达。Lentivirus-Survivin可高效转染反分化早期和晚期椎间盘髓核细胞,并促进细胞中Survivin表达。对于反分化早期椎间盘髓核细胞,Lentivirus-Survivin可增强髓核细胞活性以及细胞外基质(II型胶原和蛋白多糖)的合成;对于反分化晚期髓核细胞,其反而抑制细胞活性以及细胞外基质的合成。结论 Lentivirus-Survivin适用于反分化早期椎间盘髓核细胞,而不适用于反分化晚期的髓核细胞。
Objective To compare biological effects of Lentivirus-Survivin on the earlier or later dedifferentiated human NP cells. Methods The human NP cells were isolated and cultured using tissue culture technique. Lentivirus transfect rate to the earlier or later dedifferentiated NP cells was detected with PEGF. The expression and biological effects of Lentivirus-Survivin were detected and compared by the immunofluence, MTT, Western-Blot or Antonopulos methods. Results Survivin protein was expressed in the earlier dedifferentiated NP cells, not in the later dedifferentiated NP cells. Lentivirus-Survivin could transfect efficiently the earlier and later dedifferentiated NP cells, and promote Survivin protein to be expressed efficiently. Lentivirus-Survivin could enhance the earlier dedifferentiated human NP cells viability and the synthesis of the glycosaminoglycan or collagen type Ⅱ, but it inhibited those of the later dedifferentiated NP cells. Conclusion Lentivirusvirus-Survivin can be used for the earlier dedifferentiated cells, but it is not applicable for the later ones.
出处
《中国骨与关节损伤杂志》
2015年第9期954-958,共5页
Chinese Journal of Bone and Joint Injury
基金
国家自然科学基金(81171758)
关键词
慢病毒
生存素
椎间盘
髓核细胞
基因治疗
Lentivirus
Survivin
Intervrtebral disc
Nucleus pulposus cells
Gene therapy
