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均显著降低（P＜0.05、0.01），除游离姜黄素组外，各给药组LDH显著降低（P＜0.05、0.01）；各给药组IL-6、TNF-α均显著降低（P＜0.01、0.05）；各给药组VEGF、Bcl-2、Akt、p-PI3K的蛋白表达均显著下调（P＜0.05、0.01），cleaved Caspase-3和P53蛋白表达均显著上调（P＜0.05、0.01）； GAMCLCL （4 mg·kg^-1）给药7 d与14 d的抗肿瘤效果相似，明显强于游离姜黄素。结论 GAMCLCL能显著增强姜黄素的肝靶向性和抗肝癌作用，有利于提高荷瘤大鼠的无进展生存期。

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 months, with no precipitation. GAMCLCL did not exhibit any aggregation or precipitation when mixed with serum. Live imaging experiments showed that fluorescence was mainly concentrated in the liver at various time points after administration. The liver fluorescence was strong at 10, 30, and 60 min, significantly weakened at 120 min, and basically disappeared at 240 min. Compared with model group, the tumor mass of rats in each treatment group was significantly reduced (P < 0.05, 0.01). The liver coefficient significantly decreased in each treatment group (P < 0.05, 0.01), while the spleen coefficient significantly decreased in the free curcumin group and GAMCLCL 4 mg·kg^?1 (7, 14 d) group (P < 0.01). The RBC and PLT counts were significantly increased (P < 0.01) and WBC counts were significantly decreased (P < 0.05, 0.01) in the groups of doxorubicin and GAMCLCL 4 mg·kg^?1 (7, 14 d). ALT and CRE of rats in each treatment group were significantly reduced (P < 0.05, 0.01). Except for the free curcumin group, LDH in each treatment group was significantly reduced (P < 0.05, 0.01). IL-6 and TNF-α in each administration group significantly decreased (P < 0.01, 0.05). The protein expression of VEGF, Bcl-2, AKT, and p-PI3K in each treatment group was significantly downregulated (P < 0.05, 0.01), while the protein expression of cleaved Caspase-3 and p53 was significantly upregulated (P < 0.05, 0.01). The anti-tumor effects of GAMCLCL (4 mg·kg^?1) administered for 7 d and 14 d were similar, significantly stronger than free curcumin. Conclusion GAMCLCL significantly enhanced the liver-targeting and anti-hepatocellular carcinoma effects of curcumin, and was beneficial to the progression-free survival of tumor-bearing rats.

R285.5

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）