表阿霉素丙二醇脂质体克服肿瘤耐药性的实验研究

Epirubicin Loaded with Propylene Glycol-liposomes Overcomes Multidrug Resistance in Breast Cancer

目的 制备高包封率与高稳定的盐酸表阿霉素丙二醇脂质体(EPI-PG-liposomes),评价其抗肿瘤效果。方法 采用注入法结合pH梯度主动载药技术制备EPI-PG-liposomes,以人乳腺癌MDA-MB-435及耐阿霉素的MDA-MB-435/ADR细胞株为细胞模型,通过MTT实验考察EPI-PG-liposomes的细胞毒性,流式细胞仪技术检测EPI-PG-liposomes的细胞摄取量;采用7,12-二甲基苯蒽(DMBA)诱导5周龄雌性SD大鼠建立乳腺癌模型,通过肿瘤体积抑制率,大鼠体重变化以及HE染色分析评价抗肿瘤效果;通过大鼠体内药物分布,考察丙二醇脂质体的靶向性。结果 本实验制备的EPI-PG-liposomes平均粒径为182nm,分散性指数为0.239,Zeta电位为-29.4mV,包封率达到(93.6±1.7)%。MTT结果显示EPI-PG-liposomes有高细胞毒;流式细胞仪检测结果EPI-PG-liposomes给药后的细胞内荧光强度要强于盐酸表阿霉素;经过27天的治疗,用EPI-PG-liposomes治疗的大鼠无体重减轻,肿瘤抑制率为86.79±3.13%,而表阿霉素溶液为67.55±3.61%。HE染色结果显示,EPI-PG-liposomes治疗后的肿瘤组织严重坏死,细胞核固缩甚至消失。体内分布实验证明EPI-PG-liposomes在肿瘤组织具有高分布。结论本实验制备的EPI-PG-liposomes具有较好的抗肿瘤效果。

OBJECTIVE Prepare Epirubicin loaded with propylene glycol-liposomes ( EPI-PG-liposomes) with high encapsulation efficiency and stability and investigate its antitumor effeet. METHODS pH gradient method combined with injection method was choiced to prepare EPI-PG-liposomes. Optical microscope was used to observe the appearance of EPI-PC-liposomes. Diameter and Zata potential value was also measured. MDA-MB 435 and MDA- MB-435/ADR cell lines were cell model. Cell Cytotoxicity was tested by MTT assay;Flow cytometry technology used to detect the uptake of EPI-PG-liposomes. DMBA was used to induct tumor rats. The target of EPI-PC-liposomes was evaluated by some index :tumor volume inhibition rate , record changes in body weight;HE staining analysis and distribution of EPI-PG-liposomes in vivo. RESULTS EPI-PG-liposomes showed a mean particle size of 182nm with a poly dispersity index of 0.239. The Zeta potential was-29. 4mV and entrapment efficiency was (93.6±1.7)% MTT assay showed that EPI-PGliposomes had stronger cell inhibition than EPI solution at the same concentration of EPI (P<0. 05);Flow cytometry test indicated that more amount of EPI was transferred into cells when exposed to EPI-PG-liposomes than to EPI solution. After 27 days of treatment,the inhibition rate of tumor volume growth in EPI- PC-liposomes group was (86. 79 ±3. 13)%,while EPI group was (67.55 ±3.61 )%;It is noticed that the bodyweight gain in the rats treated EPI-PG-liposomes was even slightly increased. Severe tissue necrosis was observed in the tumors from rats treated with EPI-PG-liposome;From the biodistribution of EPI-PG-liposomes in vivo, EPI-PG-liposomes can not only keep the nature of liver-targeting character, but also enhance tumor uptake efficiency. CONCLUSION EPI-PC-liposomes prepared in this study had a good advantage of antitumor.