摘要
目的:探讨PVP预处理细胞对TAT穿膜效率的影响及其作为促渗剂的安全性,以优化TAT穿膜条件。方法:人工合成荧光标记短肽TAT-FITC和作为阴性对照的无意义肽NCO-FITC作用于PVP预处理各种细胞株,荧光显微镜观察TAT-FITC的穿膜效率及其定位;荧光酶标仪定量检测不同预处理的各种细胞摄取荧光标记短肽;MTT检测PVP对各种细胞的存活率及溶血实验检测其对细胞膜结构完整性的影响。结果:经PVP预处理细胞后,TAT-FITC可有效穿膜进入细胞内,MTT结果显示适当浓度的PVP对细胞活力几乎没有影响;溶血实验也证实,PVP不影响红细胞的细胞膜完整性。结论:本实验观察显示,PVP可以明显提高TAT对培养细胞的穿膜效率,但对细胞活力影响很小。
Objective：To study the effects of pretreatment of cells with PVP on the cell membrane penetration efficiency of TAT,thus to optimize TAT penetrating conditions.Methods：Different cell lines pre-treated by PVP were incubated with FITC-labeled peptides TAT or NCO.The penetrating efficiency and the intracellular distribution of TAT were observed under fluorescence microscopy.Fluorescence intensity in different cells was quantified by fluorescence spectrum analysis.MTT and hemolysis assays were performed to evaluate the cells viability and the integrity of cell membrane.Results：Pretreatment of cells by PVP significantly enhanced the penetrating efficiency of TAT.The result of MTT showed that PVP had very little toxic effects on cell viability,and the hemolysis analysis indicated that PVP did not destroy the cell membrane.Conclusion：Pretreatment of cells with PVP can significantly improve the penetrating efficiency of TAT and has very little effect on cell viability.
出处
《中国新药杂志》
CAS
CSCD
北大核心
2012年第7期784-787,822,共5页
Chinese Journal of New Drugs
基金
湖北省自然科学基金项目(2010CDB10705)
