甘草酸替代胆固醇构建载雷公藤甲素的脂质纳米粒的适宜性研究  

Study on suitability to construct lipid nanoparticles loaded with triptolide using glycyrrhizic acid as alternative of cholesterol

作　　者：徐资怡 陈佳美 钟雪梅 吴亿晗 章津铭[1] XU Zi-yi;CHEN Jia-mei;ZHONG Xue-mei;WU Yi-han;ZHANG Jin-ming(Key Laboratory of Standardization of Chinese Materia Medica,Ministry of Education,State Key Laboratory of Southwest Specialty Chinese Medicine Resources,College of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都中医药大学药学院,西南特色中药资源国家重点实验室,中药材标准化教育部重点实验室,四川成都611137

出　　处：《中草药》2023年第3期722-733,共12页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金面上项目(81973662);国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-D-202209)。

摘　　要：目的鉴于胆固醇潜在的健康危害,探索利用甘草酸替代胆固醇,制备用以替代胆固醇脂质纳米粒(cholesterol lipid nanoparticles,CLN)的甘草酸脂质纳米粒(glycyrrhizic acid lipid nanoparticles,GLN),通过包载抗肿瘤药物雷公藤甲素(triptolide,Tr)制备载雷公藤甲素的甘草酸脂质纳米粒(GLN loaded with triptolide,Tr@GLN),探讨甘草酸替代胆固醇制备抗肿瘤纳米载体的适宜性。方法采用乙醇注入法分别制备Tr@GLN和载雷公藤甲素的胆固醇脂质纳米粒(CLN loaded with triptolide,Tr@CLN),分别测定两者粒径、多分散指数(polydispersity index,PDI)、ζ电位、药物包封率和载药量;透射电子显微镜(transmission electron microscope,TEM)拍摄其形态;共聚焦显微镜拍摄人肝癌HepG2细胞对两者摄取情况;建立皮下荷瘤肝癌小鼠模型,对比研究Tr@GLN与Tr@CLN的体内抗肿瘤效果。结果与Tr@CLN相比,Tr@GLN在粒径、PDI、ζ电位、包封率和载药量等理化性质均无显著性差异。Tr@GLN和Tr@CLN的平均粒径分别为(115.59±1.23)、(97.28±0.95)nm;ζ电位分别为(-19.63±3.14)、(-7.77±0.12)mV;Tr包封率分别为(89.70±0.39)%、(87.39±0.37)%;Tr载药量分别为(2.25±0.01)%、(2.31±0.01)%;甘草酸包封率为(87.46±0.65)%;甘草酸载药量为(13.41±0.09)%。TEM观察到,相比Tr@CLN,Tr@GLN呈更加均匀圆整的球状。GLN增加了HepG2细胞对药物的摄取。给药12 d后,Tr@GLN对荷瘤小鼠肿瘤体积抑制率达到78.9%,显著高于Tr@CLN组抑制率(42.4%)和Tr组抑制率(18.3%)(P<0.05),Tr@GLN治疗后肿瘤中重组Ki-67蛋白(recombinant Ki-67 protein,Ki-67)表达减少,血管内皮生长因子(vascular endothelial growth factor,VEGF)表达减少,并将肿瘤相关的巨噬细胞从肿瘤M2表型重新诱导为M1表型。结论制备的Tr@GLN与Tr@CLN具有相似理化性质,但GLN包载更有利于增加肿瘤细胞的摄取,Tr@GLN表现出更强的抗肿瘤作用,其抗肿瘤作用可能与抑制肿瘤细胞分化、减少肿瘤新血管生成Objective In view of the potential health hazards of cholesterol,glycyrrhizic acid was explored to replace cholesterol to prepare glycyrrhizic acid lipid nanoparticles(GLN)for replacing cholesterol lipid nanoparticles(CLN).GLN loaded with triptolide(Tr@GLN)were prepared by encapsulating the anti-tumor drug triptolide(Tr),to explore the suitability of glycyrrhizic acid replacing cholesterol in the preparation of anti-tumor nanocarriers.Methods Tr@GLN and CLN loaded with triptolide(Tr@CLN)were prepared by ethanol injection method,and the particle size,PDI,ζpotential,drug entrapment efficiency and drug loading efficiency of Tr@GLN and Tr@CLN were measured respectively.The morphology of Tr@GLN and Tr@CLN was photographed by transmission electron microscope(TEM).The cell uptake of the both in HepG2 hepatocellular carcinoma cells was measured by confocal microscopy.The subcutaneous tumor-bearing liver cancer mouse model was established,and the in vivo antitumor effects of Tr@GLN and Tr@CLN were compared.Results Compared with Tr@CLN,the physicochemical properties of Tr@GLN,such as particle size,PDI,ζpotential,entrapment efficiency and drug loading efficiency,were not significantly different.The particle sizes of Tr@GLN and Tr@CLN were(115.59±1.23),(97.28±0.95)nm;ζPotential were(-19.63±3.14),(-7.77±0.12)mV;The entrapment efficiency of Tr were(89.70±0.39)%,(87.39±0.37%)%;The drug loading efficiency of Tr were(2.25±0.01)%,(2.31±0.01)%;The entrapment efficiency of GL was(87.46±0.65)%;The drug loading efficiency of GL was(13.41±0.09)%.Compared with Tr@CLN,Tr@GLN showed a more uniform spherical shape by TEM.GLN increased drug uptake by HepG2 hepatoma cells.After 12 d of administration,the tumor volume inhibition rate of Tr@GLN on tumor-bearing mice reached 78.9%,which was significantly higher than that of Tr@CLN group(42.4%)and Tr group(18.3%)(P<0.05).The expression of recombinant Ki-67 protein(Ki-67)and vascular endothelial growth factor(VEGF)in tumors decreased after Tr@GLN treatment,and tumor-associated macrop

关 键 词：甘草酸 雷公藤甲素 胆固醇 脂质体 抗肿瘤 肿瘤免疫 

分 类 号：R283.6[医药卫生—中药学]