透明质酸与香叶醇缀合物的合成及其抗肝癌活性  

Synthesis and Anti-Hepatoma Activity of Conjugate of Hyaluronic Acid and Geraniol

作　　者：夏一帆 崔杰 黄方 张文典 段少峰 XIA Yifan;CUI Jie;HUANG Fang;ZHANG Wendian;DUAN Shaofeng(School of Pharmacy,Henan University,Kaifeng 475001,China;Department of Pharmacy,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China)

机构地区：[1]河南大学药学院,河南开封457001 [2]广东省人民医院(广东省医学科学院)药学部,广州510080

出　　处：《中国药学杂志》2023年第17期1589-1596,共8页Chinese Pharmaceutical Journal

基　　金：河南省医学科技攻关计划联合共建项目资助(2018020306);开封市科技发展计划项目资助(1903024)。

摘　　要：目的合成透明质酸(hyaluronic acid,HA)与香叶醇(geraniol,GER)缀合物,并初步评价其体外抗HepG2肝癌细胞活性。方法通过化学接枝合成了一种透明质酸-香叶醇缀合物(HA-GER)。利用核磁共振氢谱(^(1)H-NMR)和傅里叶红外光谱(FT-IR)对其结构进行确认,利用激光粒度电位仪检测了其粒径与电位并通过透射电镜(TEM)进行形貌表征。采用CCK-8染色法检测了HA-GER对肝癌细胞HepG2的细胞毒性,EdU染色法考察了其对HepG2细胞体外增殖的影响。通过Transwell小室与Invasion小室考察了其对HepG2细胞迁移和侵袭能力的影响。通过TUNEL染色法考察了其对HepG2细胞体外凋亡的影响。结果1H-NMR结果显示,HA-GER被成功合成,且GER在HA上的接枝率约为(50±5)%;FT-IR结果进一步验证了HA-GER结构;激光粒度电位仪测得其粒径为(188.4±9.7)nm,Zeta电位为-(8.53±0.63)mV,TEM显示其形貌为均一球形。CCK-8结果表明,200μmol·L^(-1)的GER工作浓度处理24h,HA-GER表现出优于GER的对HepG2细胞活性的抑制能力。HepG2的24hEDU、Transwell以及Invasion和TUNEL的实验结果显示,与透明质酸的结合提高了GER抑制HepG2细胞的增殖、迁移和侵袭,促进凋亡的能力(P<0.001)。结论将憎水的香叶醇化学接枝透明质酸,得到亲水的HA-GER,提供了增加香叶醇溶解性与可给药性的新思路。OBJECTIVE To synthesize the conjugate of hyaluronic acid(HA)and geraniol(GER)and evaluate its in vitro anticancer effect on hepatocellular carcinoma.METHODS A hyaluronic acid-geraniol conjugate(HA-GER)was synthesized by attaching HA to GER by chemical grafting.The structure of the conjugate was confirmed by hydrogen nuclear magnetic resonance(1H-NMR)and Fourier transform infrared(FT-IR)spectra.The particle size and Zeta potential were detected by a laser particle size potentiometer and the morphology was characterized by transmission electron microscopy.The cytotoxicity of HA-GER on HepG2 cells was detected by CCK-8 staining,and the effects of HA-GER on the proliferation of HepG2 cells in vitro were examined by EdU staining.The effects of HA-GER on the migration and invasion of HepG2 cells were examined in transwell and invasion compartments.The effect of HA-GER on the apoptosis of HepG2 cells in vitro was investigated by TUNEL staining.RESULTS 1H-NMR data showed that the HA-GER conjugate was successfully synthesized and the grafting ratio of GER on HA was about(50±5)%.The structure of HA-GER was further verified by FT-IR results.Its particle size was(188.4±9.7)nm,and the Zeta potential was-(8.53±0.63)mV.The TEM showed that the structure was uniform and spherical.The results of CCK-8 showed that after treatment with 200μmol·L^(-1)GER for 24 hours,HA-GER showed an extremely superior inhibitory ability to GER on HepG2 cell activity.The 24-hour experiments of EDU,transwell,invasion and TUNEL of HepG2 showed that the conjugation with hyaluronic acid improved GER's ability to inhibit the proliferation,migration and invasion of HepG2 cells and promote apoptosis(P<0.001).CONCLUSION The hydrophobic geraniol was grafted onto hyaluronic acid by chemical reaction to obtain a hydrophilic conjugate HA-GER,which provides a new idea to increase the solubility and donating property of geraniol.

关 键 词：透明质酸 香叶醇 缀合物 HEPG2肝癌细胞 抗肿瘤活性 

分 类 号：R944[医药卫生—药剂学] R965[医药卫生—药学]