越橘叶黄酮脂质体制备及对HepG2细胞的影响  

Preparation of flavonoid liposomes from lingonberry leaves and their inhibitory effect on growth of HepG2 cells

作　　者：成圆 丁淼 王思宇 樊梓鸾 CHENG Yuan;DING Miao;WANG Siyu;FAN Ziluan(College of Life Sciences,Northeast Forestry University,Harbin 150040,Heilongjiang,China;Key Laboratory of Forest Food Resources Utilization of Heilongjiang Province,Harbin 150040,Heilongjiang,China)

机构地区：[1]东北林业大学生命科学学院,黑龙江哈尔滨150040 [2]黑龙江省森林食品资源利用重点实验室,黑龙江哈尔滨150040

出　　处：《精细化工》2024年第9期1996-2005,2030,共11页Fine Chemicals

基　　金：国家自然科学基金项目(31170510);黑龙江省自然科学基金联合引导项目(LH2020C035);中央高校基本科研业务费专项(2572019BA09);黑龙江省高等教育教学改革项目(SJGY20200018);中国博士后科学基金项目(2016M600239)。

摘　　要：对红豆越橘叶采用超声辅助提取和AB-8型大孔树脂纯化获得了黄酮含量高的馏分(L6),经HPLC-MS分析越橘叶黄酮为主要成分。以L6为芯材,采用乙醇注入法制备了脂质体L6-Lips,经不同质量分数羧甲基壳聚糖(CMCS)水溶液修饰制备了CMCS-L6-Lips。通过FTIR、粒径分析、Zeta电位和透析法对CMCS-L6-Lips进行了表征,探究其在不同pH下体外释放能力及对HepG2细胞抑制能力。结果表明,L6的黄酮含量可达899.44 mg/g,主要含有芦丁、槲皮素、槲皮素-3-O-葡萄糖醛酸苷、槲皮素3-O-鼠李糖苷-7-O-葡萄糖苷、圣草酚-7-O-葡萄糖苷、儿茶素和绿原酸7种成分;CMCS溶液质量分数与CMCS-L6-Lips粒径、包封率、Zeta电位具有相关性。添加质量分数为0.4%CMCS水溶液时脂质体表面修饰效果最佳,此时,Zeta电位为–35.65 mV,表明CMCS成功附着在L6-Lips表面,且CMCS-L6-Lips颗粒之间的排斥力明显更强,稳定性更高。CMCS-L6-Lips具有pH敏感性,在pH=4.0时体外释放能力最高,动力学方程符合Weibull模型,为骨架溶蚀释放;CMCS-L6-Lips可有效抑制HepG2细胞增殖,减缓细胞迁移能力并阻滞细胞停留在S期和G2期。脂质体包封有效提高了越橘叶黄酮的溶解性,使细胞对药物摄取的能力提升,而CMCS的修饰增加了脂质体的靶向能力,有利于治疗肿瘤。The highest flavonoid fraction(L6)in lingonberry leaves was obtained from ultrasound-assisted extraction and purification with AB-8 macroporous resin,with its main components analyzed by HPLC-MS.Carboxymethyl chitosan-L6-liposome(CMCS-L6-Lips)were then prepared from CMCS aqueous solution with different mass fraction modification of L6-liposomes,which were synthesized using L6 as core material via ethanol injection method,and characterized by FTIR,particle size analysis,Zeta potential and dialysis method.Its in vitro release ability at different pH and inhibitory ability on HepG2 cells were further analyzed and evaluated.The results showed that the flavonoid content of L6 could reach 899.44 mg/g,mainly containing 7 components such as rutin,quercetin,quercetin 3-O-β-D-glucuronide,quercetin 3-O-rhamnosid-7-O-glucoside,eriodictyol 7-O-glucoside,catechin and chlorogenic acid.The mass fraction of CMCS was correlated with the particle size,encapsulation rate,and Zeta potential of CMCS-L6-Lips.The surface modification effect of liposomes was optimal when 0.4%(mass fraction)CMCS solution was added,with a Zeta potential of–35.65 mV,indicating successful attachment of CMCS to the surface of L6-Lips and stronger repulsion forces between CMCS-L6-Lips particles,resulting in higher stability.The CMCS-L6-Lips exhibited pH sensitivity,with the highest in vitro release capacity at pH=4.0.The kinetic equation conformed to the Weibull model,indicating a skeleton erosion release mechanism.CMCS-L6-Lips could effectively inhibit the proliferation of HepG2 cells,slow down cell migration,and block cells from staying in the S and G2 phases.The encapsulation of liposomes effectively improved the solubility of lingonberry leaves flavonoid,enhancing the cell's ability to uptake the drug.Furthermore,the modification of CMCS increased the targeting ability of liposomes,which was beneficial for tumor treatment.

关 键 词：越橘叶 黄酮 HPLC-MS 脂质体 pH敏感型 HEPG2细胞 动力学方程 活性跟踪法 医药原料 

分 类 号：TQ630[化学工程—精细化工] R285[医药卫生—中药学]