摘要
目的制备Legumain酶和线粒体双级靶向去氢骆驼蓬碱(HM)脂质体(KA@HM-LPS),并对其制剂学特性、体外抗肿瘤作用及生物相容性进行初步评价。方法首先,筛选KA@HM-LPS的制备方法和均质化方法,并对制备的脂质体进行表征。其次,分别测定KA@HM-LPS的血清稳定性、体外释放率、溶血百分数,以及空白脂质体作用下的细胞存活率。最后,分别测定KA@HM-LPS作用下的细胞存活率、线粒体靶向性和对肝癌细胞迁移和侵袭的抑制作用。结果选择薄膜分散法制备KA@HM-LPS,其包封率为(90.50±0.62)%;选择挤出法作为KA@HM-LPS的均质化方法。所制备的KA@HM-LPS粒径、多分散系数、Zeta电位分别为(211.40±11.67)nm、0.316±0.014和(-14.20±0.49)mV。在37℃、10%胎牛血清中,12 h后KA@HM-LPS的粒径基本稳定;KA@HM-LPS在20%血浆中的体外释放曲线符合Weibull分布,具有缓释效果;当HM质量浓度为160μg/mL时,KA@HM-LPS的溶血百分数为(4.23±0.19)%,远小于游离HM,具有安全性。当空白脂质体的质量浓度达400μg/mL时,LO2细胞的存活率为(94.40±6.12)%,载体生物相容性较好。体外细胞实验结果显示,KA@HM-LPS对Legumain酶过表达的肝癌细胞LGMN+-SK-Hep-1的抑制作用显著高于对正常肝癌细胞SK-Hep-1的抑制作用;且与SK-Hep-1细胞相比,LGMN+-SK-Hep-1细胞对载体的摄取效率更高;KA@HM-LPS可以更明显地抑制LGMN+-SK-Hep-1细胞的迁移和侵袭。结论成功制备KA@HM-LPS;该脂质体可以有效抑制Legumain酶过表达的肝癌细胞的迁移和侵袭,并提高HM的血液相容性。
OBJECTIVE To prepare Legumain enzyme and mitochondrial double-stage targeted harmine(HM)liposome(KA@HM-LPS)and preliminary evaluate its pharmaceutical properties,in vitro antitumor effect and biocompatibility.METHODS Firstly,the preparation and homogenization methods of KA@HM-LPS was screened,and prepared liposomes were characterized.Secondly,the serum stability,in vitro release rate,hemolysis percentage of KA@HM-LPS and cell survival rate under KA@BLPS were determines respectively.Finally,the cell surivival rate,mitochondrial targeting and inhibitory effects on cell migration and invasion of KA@HM-LPS were determined.RESULTS KA@HM-LPS was prepared by the thin-film dispersion method,with encapsulation efficiency of(90.50±0.62)%.The extrusion moulding method was selected as homogenization method of KA@HM-LPS.The particle size,polydispersity index,and Zeta potential of KA@HM-LPS were(211.40±11.67)nm,0.316±0.014 and(-14.20±0.49)mV,respectively.In 37℃,10%FBS,the particle size of KA@HM-LPS kept stable after 12 h.In vitro release curve of KA@HM-LPS in 20%plasma conformed to Weibull distribution and had the property of sustained release.When HM concentration was 160μg/mL,the hemolysis percentage of KA@HM-LPS was(4.23±0.19)%,which was much lower than that of free HM,with safety.When the mass concentration of KA@BLPS reaches 400μg/mL,the survival rate of LO2 cells was(94.40±6.12)%,and the biocompatibility was good.Cell test results in vitro showed that,inhibitory effect of KA@HM-LPS on liver cancer cells with overexpression of Legumain enzyme(LGMN+-SK-Hep-1)was significantly higher than that of normal liver cancer cells SK-Hep-1;compared with SK-Hep-1,LGMN+-SK-Hep-1 cells had a higher uptake efficiency of the liposome;KA@HM-LPS could significantly inhibit the migration and invasion of LGMN+-SK-Hep-1 cells.CONCLUSIONS KA@HM-LPS is prepared successfully,which can effectively inhibit the migration and invasion of liver cancer cells with Legumain enzyme overexpression,and improve the blood compatibility of HM.
作者
伊帕尔古丽·阿皮孜
贺宏吉
王昭志
李喆喆
卡迪热娅·艾克拉木
白静雅
王梅
Ipargul·Hafiz;HE Hongji;WANG Zhaozhi;LI Zhezhe;Kadirya·Akram;BAI Jingya;WANG Mei(College of Pharmacy,Xinjiang Medical University,Urumqi 830017,China)
出处
《中国药房》
CAS
北大核心
2022年第13期1565-1572,共8页
China Pharmacy
基金
国家自然科学基金资助项目(No.81760637)
新疆维吾尔自治区天山英才计划第三期资助项目
新疆天然活性组分和释药技术重点实验室项目(No.XJDX1713)
新疆医科大学大学生创新训练计划项目(No.CX2021048)。
