去氢骆驼蓬碱光敏性脂质体的构建及其体外抗宫颈癌侵袭作用研究

Preparation and effect against cervical cancer invasion in vitro of harmine-loaded photosensitive liposomes

尽管HPV疫苗及筛查方案的逐渐成熟,但宫颈癌仍然在严重威胁女性生命健康。早期宫颈癌的治疗以手术为主,拟通过热疗增强化疗作用,保留患者宫颈部位完整,提高治疗效果,为宫颈癌早期治疗提供更多治疗依据。采用薄膜分散法制备同时包载去氢骆驼蓬碱(HM)、染料IR-780的光敏性脂质体,筛选最佳条件是磷脂胆固醇膜材质量比为8∶1,药脂质量比为1∶20;通过Sephadex G-50葡聚糖凝胶柱进行分离,采用HPLC测定去氢骆驼蓬碱包封率为55.6%±0.18%;在透射电镜下观察形态圆整,分布均匀;马尔文粒度仪测定粒径为(125.2±0.62)nm及Zeta电位为(-2.55±0.76)mV;光热转换效率是光热剂的重要属性,经测定光热转换效率为27.1%±0.86%;在4℃的贮存条件下,在前14 d的包封率较稳定,无絮凝现象;并采用磺酰罗丹明B蛋白(SRB)法考察脂质体对HeLa细胞在近红外照射下的抗增殖作用比无红外照射作用强;采用Transwell实验考察脂质体对HeLa细胞的抑制侵袭迁移作用,明显抑制HeLa细胞的体外迁移和侵袭的能力。该研究为去氢骆驼蓬碱治疗宫颈癌提供更多研究基础。

Despite the development of HPV vaccines and screening programs,cervical cancer is still a serious threat to women′s health.Early-stage cervical cancer is mainly treated by surgery.However,considering the serious complications after surgery,hyperthermia is recommended to enhance the effect of chemotherapy,retain the integrity of cervix,improve the treatment effect,which provides a therapeutic basis for the early treatment of cervical cancer.The photosensitive liposomes containing harmine and dye IR-780 were prepared by thin-film dispersion method and separated by Sephadex G-50 dextran gel column.The preparation conditions were optimized as the mass ratio of phospholipid to cholesterol membrane material being 8∶1 and that of drug to lipid being 1∶20.The results of HPLC showed that the encapsulation efficiency of harmine was 55.6%±0.18%.The prepared photosensitive liposomes were round and evenly distributed under transmission electron microscope,with the particle size of(125.2±0.62)nm determined by Marvin particle size analyzer and the Zeta potential of(-2.55±0.76)mV.Additionally,the photosensitive liposomes had the photothermal conversion efficiency,an important property of photothermal agent,of 27.1%±0.86%.The photosensitive liposomes stored at 4℃showed stable encapsulation efficiency in the first 14 days without flocculation.The sulforhodamine B(SRB)assay was employed to determine the inhibitory effect of the liposomes on the proliferation of HeLa cells under near-infrared(NIR)irradiation or not,which showcased stronger inhibitory effect under NIR irradiation.The results of Transwell assay indicated that the prepared liposomes significantly inhibited the invasion and migration of HeLa cells in vitro.The findings of this study provide a basis for the treatment of cervical cancer with harmine.