阿霉素及棉酚双药纳米载体对U87胶质瘤细胞的协同抑制作用

Synergistic inhibitory characteristics of doxorubicin and gossypol dual-drugs nanocarriers for U87 glioma cell

目的:建立一种具有协同治疗效果的阿霉素及棉酚双药纳米载体,并检测其协同抗神经胶质瘤的效果。方法:通过MTT法检测阿霉素和棉酚对人脑胶质瘤细胞U87的协同抑制作用,使用透明质酸和阳离子两亲性淀粉为材料构建双药纳米载体,并对其性能进行表征,利用荧光标记的纳米载体观察其在U87细胞和体内模型上的靶向递送水平,最后检测双药纳米载体体内抗肿瘤效果。结果:阿霉素和棉酚针对U87细胞有显著的协同抑制效果,所制备的双药纳米载体粒径为145.7 nm±5.6 nm,棉酚的包裹率为93.8%±2.5%载药量18.2%±2.1%;盐酸阿霉素的包裹率为89.4%±4.7%载药量1.9%±0.6%,72 h阿霉素释放率小于40%,低于40℃环境下稳定时间超过5周。药纳米载体可在1 h内即可在U87细胞中达到较高的积累量,并持续释放携带的药物,细胞抑制效果与游离双药一致,体内实验表明,双药纳米载体可有效靶向肿瘤组织,体内抑瘤效果显著,抑瘤率高于90%,明显好于游离给药组,且毒性降低。结论:阿霉素与棉酚双药纳米载体可将两种药物有效递送至肿瘤组织和细胞内,并发挥出显著的协同抑瘤作用。

Objective： The preparation of a kind of synergistic anti-glioma nanocarriers,which are loading doxorubicin and gossypol,and research of the antitumor effect. Methods： MTT method was used to test synergistic inhibition of doxorubicin and gossypol against glioma cell lines U87. Cationic amphipathic starch and hyaluronan were used in coencapsulations of those two drugs. Then properties of the nanocarriers were measured. The cellular and in vivo delivery of the samples was observed via fluorescence-loaded nanocarriers. U87 cell line and the tumor-bearing nude mice models were used in tumor inhibition test. Results： Doxorubicin and gossypol had significant coordinate inhibition in U87 cell. The size and zeta potential of the dual drug nanocarriers were 145. 7 nm ± 5. 6 nm. The nanocarriers showed very effective capacities in encapsulations of drugs,encapsulation efficiencies of gossypol and doxorubicin were 93. 8% ±2. 5% and 89. 4% ± 4. 7% with respect to drug-loading capacities of 18. 2% ± 2. 1% and 1. 9% ± 0. 6%. Less than40% of doxorubicin was released within 72 h,and the sizes were very stable when the temperature was below 40℃ within 5 weeks. The nanocarriers could deliver the drugs into cytoplasm within 1 h,and sustained release payloads. Furthermore,it had similar cytotoxicities compared to free dual-drug treatment. In vivo near-infrared fluorescence imagesshowed that the nanocarriers efficiently increased accumulation in tumor tissues. Above all the nanocarriers exhibited much more effective anti-tumor potency,the tumor inhibitory rate was more than 90%,and meanwhile,toxicity was lower than free drugs treatments. Conclusion： Dual-drugs nanocarriers were able to co-deliver doxorubicin and gossypol into tumor tissues and cells and exhibited significant synergistic anti-glioma effect.