转铁蛋白修饰辣蓼黄酮脂质体的制备和体外释放研究  

Preparation and In Vitro Release Study of Transferrin-Modified Flavonoid of Polygonum hydropiper L.Liposomes

作　　者：吴昌旭 张金武 彭春子 吕茂杰 梁静 蒋凯屹 赖鹏羽 韦英益[1] 陈海兰[1] 何家康[1] 胡庭俊[1] 于美玲 WU Changxu;ZHANG Jinwu;PENG Chunzi;LYU Maojie;LIANG Jing;JIANG Kaiyi;LAI Pengyu;WEI Yingyi;CHEN Hailan;HE Jiakang;HU Tingjun;YU Meiling(College of Animal Science and Technology,Guangxi University,Nanning 530004,China)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004

出　　处：《中国兽医杂志》2025年第3期1-11,共11页Chinese Journal of Veterinary Medicine

基　　金：国家自然科学基金项目(32202850,32360894);广西自然科学基金面上基金项目(2021GXNSFAA196004);广西大学自然科学与技术创新发展倍增计划项目(2024BZRC013)。

摘　　要：为优化转铁蛋白(Tf)修饰的辣蓼黄酮(FPHL)脂质体(Tf-FPHL-Lip)的最佳制备工艺,明确其表征和体外释放特性,本试验首先利用乙醇注入法、薄膜分散法和逆向蒸发法制备辣蓼黄酮脂质体(FPHL-Lip),优选出包封率最高的制备方法。以包封率为考察指标,通过单因素试验和Box-Behnken响应面法优化FPHL-Lip制备工艺;利用二硬脂酰磷脂酰乙醇胺-聚乙二醇2000-羧酸(DSPE-PEG2000-COOH)上的活性末端羧基将Tf和脂质体在催化剂作用下连接,通过单因素试验优化Tf-FPHL-Lip的制备工艺,并对其形态、粒径、多分散指数(PDI)、Zeta电位和体外释放情况进行考察。结果显示,薄膜分散法制备Tf-FPHL-Lip的最优条件是磷胆比为6.28∶1、药脂比为1∶6.48、制备温度为58.48℃、水化体积为15 mL、表面活性剂浓度为7.5 mg/mL、超声时间为6 min、二硬脂酰基磷脂酰乙醇胺-聚乙二醇2000(DSPE-MPEG2000)与磷脂的摩尔比为6%、DSPE-PEG2000-COOH与DSPE-MPEG2000的摩尔比为5%、1-乙基-(3-二甲基氨基丙基)碳二亚胺盐酸盐(EDC)、N-羟基琥珀酰亚胺(NHS)与DSPE-PEG2000-COOH的摩尔比为10∶10∶1,Tf与DSPE-PEG2000-COOH的摩尔比为1∶40;在优化条件下制备的Tf-FPHL-Lip平均包封率为(63.35±3.41)%,平均粒径为(96.46±1.32)nm,平均PDI为0.312±0.029,平均Zeta电位为(-2.02±0.22)mV,平均连接率为(68.64±0.07)%,眼观为淡黄色、澄清、透明的液体,在电镜下为单层脂质体,呈球形,表面完整,粒径约100 nm;相对于FPHL,Tf-FPHL-Lip在体外具有明显的缓释作用。结果表明,优化的制备条件稳定可行,所制备的Tf-FPHL-Lip包封率较高,粒径小且均匀,为Tf-FPHL-Lip的进一步开发利用提供理论基础。In order to optimize the preparation process of transferrin(Tf)-modified flavonoid of Polygonum hydropiper L.(FPHL)liposomes(Tf-FPHL-Lip),and to clarify their characterization and in vitro release properties,this study first prepared FPHL liposomes(FPHL-Lip)using ethanol injection,thin-film dispersion,and reverse evaporation methods,and selected the method with the highest encapsulation efficiency.The preparation process of FPHL-Lip was optimized by evaluating the encapsulation efficiency as the key indicator through single-factor experiments and Box-Behnken response surface methodology.The Tf was connected to the liposomes using the active terminal carboxyl group of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[(polyethylene glycol)-2000]-carboxylic acid(DSPE-PEG2000-COOH)under a catalytic condition.The preparation process of Tf-FPHL-Lip was further optimized by single-factor experiments,and the morphology,particle size,polydispersity index(PDI),Zeta potential,and in vitro release were evaluated.The results showed that the optimal conditions for preparing Tf-FPHL-Lip using the thin-film dispersion method were a lipid-to-cholesterol ratio of 6.28˸1,a drug-to-lipid ratio of 1˸6.48,a preparation temperature of 58.48℃,a hydration volume of 15 mL,a surfactant concentration of 7.5 mg/mL,an sonication time of 6 minutes,a N-(carbonyl-methoxypolyethylene glycol 2000)-1,2-distearoyl-sn-glycerol-3-phosphoethanolamine,sodium salt(DSPE-MPEG2000)-to-phospholipid molar ratio of 6%,a DSPE-PEG2000-COOH-to-DSPE-MPEG2000 molar ratio of 5%,an 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(EDC)-N-Hydroxysuccinimide(NHS)-DSPE-PEG2000-COOH molar ratio of 10˸10˸1,and a Tf-to-DSPE-PEG2000-COOH molar ratio of 1˸40.Under these optimized conditions,the prepared Tf-FPHL-Lip showed an average encapsulation efficiency of(63.35±3.41)%,an average particle size of(96.46±1.32)nm,an average PDI of 0.312±0.029,an average Zeta potential of(-2.02±0.22)mV,and an average conjugation rate of(68.64±0.07)%.The appearance was

关 键 词：辣蓼黄酮 转铁蛋白 脂质体 响应面法 表征 体外释放 

分 类 号：S859.53[农业科学—临床兽医学]