载阿霉素纳米粒温敏凝胶复合体系的体内缓释性能和抗肿瘤作用及瘤内滞留性能评价  

Evaluation of doxorubicin-containing nanoparticle thermosensitive gel composite system on its extended release in vivo,antitumor,and intratumor retention

作　　者：杜华康 陆苑[1] 金阳 陈玉颖 王永林 李勇军[2,3] 刘文 DU Huakang;LU Yuan;JIN Yang;CHEN Yuying;WANG Yonglin;LI Yongjun;LIU Wen(Guizhou Provincial Key Laboratory of Pharmaceutics&State Key Laboratory of Functions and Applications of Medicinal Plants,Guizhou Medical University,Guiyang 550004,Guizhou,China;School of Pharmacy,Guizhou Medical University,Guiyang 550004,Guizhou,China;Engineering Research Center for the Development and Application of Ethnic Medicine and TCM of Ministry of Education,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学贵州省药物制剂重点实验室&省部共建药用植物功效与利用国家重点实验室,贵州贵阳550004 [2]贵州医科大学药学院,贵州贵阳550004 [3]贵州医科大学民族药与中药开发应用教育部工程研究中心,贵州贵阳550004

出　　处：《贵州医科大学学报》2024年第3期361-367,374,共8页Journal of Guizhou Medical University

基　　金：中央引导地方科技专项项目(黔科中引地〔2018〕4006);贵州省科技计划项目(黔科合平台人才〔2016〕5613,5677)。

摘　　要：目的探讨载阿霉素(DOX)纳米粒温敏凝胶(DOX-NPs-Gel)复合体系的体内缓释性能、抗肿瘤作用及瘤内滞留性能。方法18只SD雄性大鼠随机均分为DOX组、DOX-碘油组(4 g/L DOX溶液1 mL与碘油1 mL混匀现配,5 mL/kg)及DOX-NPs-Gel组(5 mg/kg DOX+2 g/L DOX),腹腔注射,分别于给药后不同时间点经眼底静脉丛穿刺取血,采用超高效液相色谱-串联质谱(UPLC-MS/MS)技术检测血浆中DOX的浓度,利用WinNonLin 8.1软件拟合主要药代参数[半衰期(t 1/2)、峰浓度(C max)、曲线下面积(AUC)、清除率(CL)、平均驻留时间(MRT)];培养、收集小鼠肝癌细胞株H22细胞制成悬液,单次接种于90只雄性ICR小鼠右腋皮下构建荷瘤模型,分2部分进行实验,第一部分取荷瘤小鼠36只均分为生理盐水组(注射用生理盐水,5 mL/kg)、空白NPs-Gel组(100 mg空白NPs冻干粉分散于1 mL空白Gel,5 mL/kg)、碘油组(碘油,5 mL/kg)、DOX组(2 g/L DOX,5 mL/kg)、DOX-碘油组(4 g/L DOX溶液1 mL与碘油1 mL混匀现配,5 mL/kg)及DOX-NPs-Gel组(2 g/L DOX,5 mL/kg),瘤内注射、给药1次,观测10 d,给药后每2天称体质量1次并计算各组小鼠的体质量变化百分比,称重后同时测量各组小鼠瘤体宽度、长度并计算肿瘤体积(V),观测结束时脱颈处死、剥离肿瘤组织,称取各组小鼠瘤体质量并计算抑瘤率,然后制作切片采用苏木精-伊红(HE)染色观察组织学特征;第二部分取荷瘤小鼠54只均分为DOX组、DOX-碘油组及DOX-NPs-Gel组,给药剂量和途径同前,给药后12、24、48、72、96及120 h时取各组3只小鼠脱颈处死,剥离肿瘤组织,采用UPLC-MS/MS检测瘤体组织中各时间点的DOX瘤内滞留率。结果体内缓释性能研究结果显示,相比于其余组,DOX-NPs-Gel组大鼠t 1/2延长、CL降低(P<0.05);抗肿瘤作用考察结果显示,与其它治疗组相比,DOX-NPs-Gel组小鼠肿瘤组织生长最慢,瘤重降低(P<0.05),抑瘤率最高(76.55%),HE染色显示肿瘤组织出现大范围凋亡和坏死情况Objective To investigate the extended release performance in vivo,anti-tumor effect and intratumor retention performance of doxorubicin-loaded nanoparticles thermosensitive gel(DOX-NPs-Gel)composite system.Methods Eighteen SD male rats were randomly divided into doxorubicin(DOX)group,DOX-iodine oil group(1 mL 4 g/L DOX solution mixed with 1 mL iodine oil on site,5 mL/kg)and DOX-NPs-Gel group(5 mg/kg DOX+2 g/L DOX);then subjects received intraperitoneal injection;blood was taken through the fundus venous plexus at different time points after administration of drugs;the concentration of DOX in plasma was detected by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS),and the main pharmacokinetic parameters were fitted by WinNonLin 8.1 software[half life period(t 1/2),C max,area under the curve(AUC),clearance(CL),mean residence time(MRT)].Rats hepatocarcinoma strain H22 was cultivated and collected for suspension;single vaccination was performed in right axilla subcutaneous part of 90 male ICR rats for load tumor.Such experiment was divided into two parts:part one,36 rats with load tumor were evenly divided into normal saline group(injected with normal saline,5 mL/kg),blank NPs-Gel group(100 mg blank NPs lyophilized powder distributed in 1 mL blank Gel,5 mL/kg),iodine oil group(iodine oil,5 mL/kg),DOX group(2 g/L DOX,5 mL/kg),DOX-iodine oil group(1 mL 4 g/L DOX mixed with 1 mL iodine oil on site,5 mL/kg)and DOX-NPs-Gel group(2 g/L DOX,5 mL/kg);intratumor injected for one time and observed for 10 d,body mass was measured every two days after drug administration and calculated changes of body mass ration of all groups;the size and length of tumor was measured after weighing and tumor volume(V)was calculated;all the mice were sacrificed by neck removal after observation finished,tumor tissue was stripped,tumor mass was weighed and tumor inhibition rate was calculated;histological features of tumor tissue was observed by slicing and HE staining.Part two,54 load tumor mice were evenly divided int

关 键 词：纳米粒 温敏凝胶 药代动力学 抗肿瘤作用 缓释作用 肝动脉化疗栓塞术 

分 类 号：R943[医药卫生—药剂学]