载紫杉醇的大黄酸偶联物胶束的体外释放及抗肿瘤研究  

In Vitro Release and Anti-Tumor Studies of Paclitaxel-Loaded Rhein Conjugate Micelles

作　　者：黄彩霞 欧阳惠枝 黄秋萍 陆伟利 王晓颖[1] 徐伟[1] HUANG Caixia;OUYANG Huizhi;HUANG Qiuping;LU Weili;WANG Xiaoying;XU Wei(College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学药学院,福州350122

出　　处：《中国药学杂志》2024年第15期1408-1415,共8页Chinese Pharmaceutical Journal

基　　金：国家自然科学基金项目资助(81603301);福建省科技厅引导性项目资助(2020Y0050)。

摘　　要：目的研究载紫杉醇的大黄酸偶联物(PTX/TPGS-CR)胶束的体外释放行为及其体外抗肿瘤效果。方法采用透析法,以泰素(Taxol?)为对照,考察PTX/TPGS-CR胶束在不同释放介质中的释药行为。采用四甲基偶氮唑蓝(MTT)法,以Taxol?为对照,评估PTX/TPGS-CR胶束对小鼠乳腺癌4T1细胞的细胞毒性。通过流式细胞仪检测PTX/TPGS-CR胶束对人宫颈癌HeLa细胞的细胞周期、凋亡的影响。通过划痕实验考察PTX/TPGS-CR胶束对HeLa、4T1细胞迁移的影响。结果体外释放实验结果显示,在pH 7.4磷酸盐缓冲液(PBS)中,PTX/TPGS-CR胶束在12和72 h内分别累积释放50.84%和71.38%;在模拟肿瘤微环境pH 5.8 PBS中,PTX/TPGS-CR胶束在12 h内累积释放79.19%,且72 h几乎完全释放。细胞毒试验结果显示,PTX/TPGS-CR胶束对4T1细胞表现出较强的肿瘤细胞增殖抑制作用,并呈时间和浓度依赖性,72 h时半抑制浓度(IC_(50))为0.3219 nmol·mL^(-1)。PTX/TPGS-CR胶束能引起HeLa、4T1细胞发生早期凋亡和晚期凋亡;可使HeLa细胞在PTX阻滞G2/M期的基础上同时增加G0/G1期阻滞,抑制细胞分裂增殖。PTX/TPGS-CR胶束能抑制HeLa、4T1细胞迁移,减少肿瘤转移。结论PTX/TPGS-CR胶束具有很好的pH响应释药特征,在血液环境中缓释药物,而在肿瘤微环境中实现快速释药;具有很好的肿瘤增殖抑制效果,促进细胞凋亡,抑制肿瘤细胞迁移,具有较好的肿瘤治疗潜力。OBJECTIVE To study the in vitro release behavior and anti-tumor effect of paclitaxel-loaded rhein conjugate(PTX/TPGS-CR)micelles.METHODS The in vitro drug release behavior of PTX/TPGS-CR micelles in different release media was investigated by dialysis method with Taxol?as the control.The cytotoxicity of PTX/TPGS-CR micelles on mouse breast cancer 4T1 cells was evaluated by MTT assay with Taxol?as a control.The effect of PTX/TPGS-CR micelles on the cell cycle and apoptosis of human cervical carcinoma HeLa cells was detected by flow cytometry.The effect of PTX/TPGS-CR micelles on the migration of HeLa and 4T1 cells was investigated by scratch experiments.RESULTS The results of in vitro release assay showed that 50.84%and 71.38%of PTX in PTX/TPGS-CR micelles were released in pH 7.4 phosphate buffer(PBS)within 12 and 72 h,respectively,79.19%of PTX in PTX/TPGS-CR micelles were released in pH 5.8 PBS which simulated the tumor microenvironment within 12 h,and almost all of PTX in the micelles was released within 72 h.The results of the cytotoxicity assay showed that PTX/TPGS-CR micelles had a strong inhibitory effect on the proliferation of 4T1 cells and displayed a time-and-concentration-dependent characteristic,with IC_(50)of 0.3219 nmol·mL~(-1)at 72 h.PTX/TPGS-CR micelles could induce early apoptosis and late apoptosis in HeLa and 4T1 cells.PTX/TPGS-CR micelles can induce HeLa cells arrest in G2/M phase(arrested by PTX)and G0/G1 phase,which inhibited cell division and proliferation.PTX/TPGS-CR micelles can inhibit the migration of HeLa and 4T1 cells and reduce tumor metastasis.CONCLUSION PTX/TPGS-CR micelles have good pH-responsive drug release characteristics,which can release drugs slowly in the blood environment and rapidly in the tumor microenvironment.PTX/TPGS-CR micelles could inhibit tumor cells proliferation and migration and promote cell apoptosis.PTX/TPGS-CR micelles have a good tumor treatment potential.

关 键 词：紫杉醇 聚合物胶束 体外释放 抗肿瘤 细胞实验 

分 类 号：R944[医药卫生—药剂学]