甘草次酸修饰的姜黄素阳离子脂质体肝靶向性和抗肝癌作用研究  

Study on anti-hepatocellular carcinoma efficacy of glycyrrhetinic acid-modified curcumin-loaded cationic liposomes

作　　者：刘炎清 李瀚旻[2] 常明向[1] 陈诺 LIU Yanqing;LI Hanmin;CHANG Mingxiang;CHEN Nuo(Hubei University of Chinese Medicine,Hubei Key Laboratory of Resources and Chemistry of Chinese Medicine,Wuhan 430065,China;Hubei Provincial Hospital of Traditional Chinese Medicine,Affiliated with Hubei University of Traditional Chinese Medicine,Wuhan 430061,China)

机构地区：[1]湖北中医药大学中药资源与中药化学重点实验室,湖北武汉430065 [2]湖北中医药大学附属湖北省中医院,湖北武汉430061

出　　处：《药物评价研究》2024年第4期754-764,共11页Drug Evaluation Research

基　　金：国家自然科学基金资助项目(81973669;81274147);广西中医药跨学科创新团队项目(GZKJ2306)。

摘　　要：目的研究甘草次酸修饰的姜黄素阳离子脂质体(GAMCLCL)肝靶向性以及抗肝癌作用。方法制备甘草次酸(GA)配体——GA和十八胺盐(SGO),用红外光谱和质谱检测;并进一步利用SGO制备GAMCLCL,透射电镜观察脂质体形态,Nano ZS90纳米粒度仪测定脂质体粒径与电位;采用活体成像系统观察GAMCLCL小鼠体内荧光分布。Wistar大鼠随机分为对照组、模型组、阿霉素(阳性药,2 mg·kg^(−1))组、姜黄素(20 mg·kg^(−1))组和GAMCLCL低、高剂量(2、4 mg·kg^(−1))组,除对照组外,采用Walker-256细胞种植法制备肝原位移植瘤模型,每天1次,尾iv给药,连续7 d;另设GAMCLCL(4 mg·kg^(−1))给药14 d组;称肿瘤质量,计算肝脏系数、脾脏系数;全自动生化分析仪测定血液红细胞(RBC)、白细胞(WBC)、血小板(PLT)、丙氨酸氨基转移酶(ALT)和肌酐(CRE)水平,试剂盒法检测乳酸脱氢酶(LDH)水平;ELISA法检测血清肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平;Western blotting法检测肿瘤组织血管内皮生长因子(VEGF)、cleaved Caspase-3、Bcl-2、p53、p-PI3K、p-Akt蛋白表达水平。结果红外光谱和质谱的结果可以验证GA与十八胺的反应生成了SGO;GAMCLCL外观呈球形,其粒径为(194±0.25)nm,聚合物分散性指数(PDI)为0.21±0.02,电位为(31.9±0.31)mV;GAMCLCL在2个月内呈黄色透明溶液,无沉淀析出;GAMCLCL与血清混合未出现任何聚集和沉淀现象;活体成像实验显示,给药后各时间点荧光主要集中在肝脏,10、30、60 min时肝脏荧光较强,120 min时肝脏荧光明显减弱,240 min时肝脏荧光基本消失。与模型组比较,各给药组大鼠肿瘤质量均明显减轻(P<0.05、0.01);各给药组肝脏系数显著降低(P<0.05、0.01),游离姜黄素组、GAMCLCL 4 mg·kg^(−1)(7、14 d)组脾脏系数显著下降(P<0.01);阿霉素及GAMCLCL 4 mg·kg^(−1)(7、14 d)组的RBC和PLT计数均显著升高(P<0.01),WBC计数均显著降低(P<0.05、0.01);各给药组大鼠ALT、CRE均Objective To investigate the liver targeting of glycyrrhizic acid-modified curcumin-loaded cationic liposomes(GAMCLC)and its anticancer effects.Methods Prepared glycyrrhetinic acid(GA)ligands-GA and octadecylamine salt(SGO)and detected them using infrared spectroscopy and mass spectrometry,and further used SGO to prepare GAMCLC,and observed the morphology of liposomes under transmission electron microscopy,and measured the particle size and potential of liposomes using Nano ZS90 nanoparticle size analyzer.Using a live imaging system to observe the fluorescence distribution in GAMCLC mice.Wistar rats were randomly divided into a control group,a model group,a doxorubicin(positive drug,2 mg·kg^(−1))group,a curcumin(20 mg·kg^(−1))group,and a GAMCLCL low and high dose(2,4 mg·kg^(−1))group.Except for the control group,a Walker-256 cell seeding method was used to prepare a liver orthotopic transplantation tumor model.The rats were administered once a day for 7 consecutive days,and set up a 14 day group of GAMCLCL(4 mg·kg^(−1))for administration.Weighed the tumor mass,calculated the liver coefficient and spleen coefficient.The levels of red blood cells(RBC),white blood cells(WBC),platelets(PLT),alanine aminotransferase(ALT),and creatinine(CRE)were measured using a fully automated biochemical analyzer,and lactate dehydrogenase(LDH)levels were detected using a reagent kit method.ELISA method was used for detecting serum tumor necrosis factor-α(TNF-α)and the level of interleukin-6(IL-6).Western blotting was used to detect the expression levels of vascular endothelial growth factor(VEGF),cleaved Caspase-3,Bcl-2,p53,p-PI3K,and p-AKT proteins in tumor tissues.Results The results of infrared spectroscopy and mass spectrometry could verify that the reaction between GA and octadecylamine generated SGO.GAMCLCL had a spherical appearance with a particle size of(194±0.25)nm,a polymer dispersibility index(PDI)of 0.21±0.02,and a potential of(31.9±0.31)mV.GAMCLCL appeared as a yellow transparent solution within two mon

关 键 词：姜黄素 甘草次酸 阳离子脂质体 Walker 256肝原位移植瘤 抗肿瘤作用 

分 类 号：R285.5[医药卫生—中药学]