载小白菊内酯靶向脂质体的制备及体外抗肿瘤研究

Preparation and anti-tumor study of parthenolide targeting liposomes in vitro

目的制备搭载环状多肽cNGQ的小白菊内酯脂质体(cNGQ-Lips@PTL)并进行处方优化以及体外抗肿瘤研究。方法以包封率为指标,用Box-Behnken设计—效应面法优化处方,用透射电镜、激光粒度仪、动态透析法、可见分光光度法等检测cNGQ-Lips@PTL的形态、粒径、体外释药行为、血清稳定性和血液相容性;用荧光显微镜、CCK-8法、凋亡检测试剂盒和流式细胞术等检测脂质体的细胞相容性、cNGQ-Lips@PTL对非小细胞肺癌细胞的靶向效率和细胞毒性。结果成功制备cNGQ-Lips@PTL,粒径均一,呈类球形态,平均粒径为128.3 nm,Zeta电位为-26.7 mV,包封率为74.89%,载药量12.08%。cNGQ-Lips@PTL在pH=5.5的PBS中,24 h累计释放量62.35%。细胞摄取实验中,cNGQ-Lips@PTL组荧光强度和摄取率明显高于PTL-Lips组以及游离组。在细胞毒性实验,cNGQ-Lips@PTL中PTL浓度为80μmol·L^(-1)时,孵育48 h后,A549细胞存活率为5.21%。在细胞凋亡实验中,cNGQ-Lips@PTL组98.74%的细胞处于凋亡晚期。结论本研究制备的cNGQ-Lips@PTL具有较为明显的靶向作用以及较强的抑制非小细胞肺癌的效果,为后续开发具有靶向作用的纳米制剂提供了一种较有价值的参考。

Objective To prepare cyclic polypeptide cNGQ-loaded parthenolide liposomes(cNGQ-Lips@PTL),optimize the formulation and perform anti-tumor study in vitro.Methods Using encapsulation rate as index,Box-Behnken design-effect surface method was used to optimize the prescription.The morphology,particle size,in vitro drug release behavior,serum stability and blood compatibility of cNGQ-Lips@PTL were detected by transmission electron microscopy,laser particle size analyzer,dynamic dialysis and visible spectrophotometry.Fluorescence microscopy,CCK-8 assay,apoptosis detection kit and flow cytometry were used to detect cell compatibility,cNGQ-Lips@PTL targeting efficiency and cytotoxicity of NSCLC cells.Results cNGQ-Lips@PTL was successfully prepared with uniform particle size and spheroid shape,the average particle size was 128.3 nm,the Zeta potential was-26.7 mV,the encapsulation rate was 74.89%,and the drug loading was 12.08%.cNGQ-Lips@PTL In PBS with pH=5.5,the cumulative release in 24 h was 62.35%.In the cell uptake experiment,the fluorescence intensity and uptake rate of cNGQ-Lips@PTL group were significantly higher than those of TL-Lips group and free group.In the cytotoxicity test,the PTL concentration in cNGQ-Lips@PTL was 80μmol[DK]·L^(-1),and the survival rate of A549 cells was 5.21%after incubation for 48 h.In the apoptosis experiment,98.74%of cells in cNGQ-Lips@PTL group were in the late stage of apoptosis.Conclusion The cNGQ-Lips@PTL prepared in this study has obvious targeting effect and strong inhibitory effect on non-small cell lung cancer,which provides a valuable reference for the subsequent development of nano preparations with targeting effect.