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花生五烯酸与透明质酸接枝物的合成及其抗肝癌活性

崔杰, 夏一帆, 张文典, 段少峰

崔杰, 夏一帆, 张文典, 段少峰. 花生五烯酸与透明质酸接枝物的合成及其抗肝癌活性[J]. 中国药科大学学报, 2022, 53(1): 46-53. DOI: 10.11665/j.issn.1000-5048.20220107
引用本文: 崔杰, 夏一帆, 张文典, 段少峰. 花生五烯酸与透明质酸接枝物的合成及其抗肝癌活性[J]. 中国药科大学学报, 2022, 53(1): 46-53. DOI: 10.11665/j.issn.1000-5048.20220107
CUI Jie, XIA Yifan, ZHANG Wendian, DUAN Shaofeng. Synthesis of eicosapentaenoic acid and hyaluronic acid graft copolymer and its anti-hepatoma activity[J]. Journal of China Pharmaceutical University, 2022, 53(1): 46-53. DOI: 10.11665/j.issn.1000-5048.20220107
Citation: CUI Jie, XIA Yifan, ZHANG Wendian, DUAN Shaofeng. Synthesis of eicosapentaenoic acid and hyaluronic acid graft copolymer and its anti-hepatoma activity[J]. Journal of China Pharmaceutical University, 2022, 53(1): 46-53. DOI: 10.11665/j.issn.1000-5048.20220107

花生五烯酸与透明质酸接枝物的合成及其抗肝癌活性

基金项目: 河南省医学科技攻关计划联合共建项目(No.2018020306);开封市科技发展计划资助项目(No.1903024)

Synthesis of eicosapentaenoic acid and hyaluronic acid graft copolymer and its anti-hepatoma activity

Funds: This study was supported by the Joint Construction Project of Henan Medical Science and Technology Research Project (No.2018020306) and the Program of Kaifeng Science and Technology Development Project (No.1903024)
  • 摘要: 合成花生五烯酸(EPA)与透明质酸(HA)耦合物,初步评价其体外抗肝癌活性。通过胱胺将花生五烯酸与透明质酸连接,合成了一种透明质酸-花生五烯酸接枝物(HA-EPA),利用核磁共振仪(1H NMR)和傅里叶变换红外光谱仪(FT-IR)对其结构进行了表征,利用激光粒度及Zeta电位分析仪检测了其粒径与电位。采用MTT法检测了HA-EPA对肝癌细胞HepG2,Huh-7和正常肝细胞LX-2的体外抗细胞增殖作用。利用EdU染色与TUNEL染色法,考察了HA-EPA对HepG2细胞体外增殖与凋亡的影响。流式细胞术进一步验证了凋亡情况。通过迁移和侵袭实验考察了HA-EPA对HepG2细胞迁移和侵袭能力的影响。1H NMR结果显示,HA-EPA被成功合成,且EPA在HA上的接枝率约(40 ± 5)%;FT-IR结果进一步确证了HA-EPA的结构;粒径为(162.5 ± 10.2) nm,电位为-(4.47 ± 0.31) mV;MTT结果表明,随着药物处理时间延长,相同EPA含量下HA-EPA表现出优于EPA的对HepG2与Huh-7细胞活性抑制能力。当作用48 h,HA-EPA对正常肝细胞LX-2的毒性小于EPA。HepG2的24 h增殖、凋亡、迁移与侵袭实验结果显示,透明质酸的接枝提高了EPA抑制HepG2细胞增殖,促进凋亡,抑制迁移与侵袭的能力(P < 0.001),说明HA的接枝可以明显增强EPA的肝癌抑制效果并起到一定的减毒作用。
    Abstract: In this study, the conjugate of eicosapentaenoic acid (EPA) and hyaluronic acid (HA) was synthesized and the anti-hepatoma activities in vitro were evaluated.The hyaluronic acid-eicosapentaenoic acid (HA-EPA)nanoparticle was synthesized by linking eicosapentaenoic acid with hyaluronic acid with cystamine.The structure of HA-EPA was characterized by nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy (FT-IR).Laser particle sizer and Zeta potential analyzer were used to detect the size and potential of HA-EPA.MTT assay was used to detect the anti-proliferative effect of HA-EPA on HepG2, Huh-7 and LX-2 cells in vitro.The effects of HA-EPA nanoparticles on the proliferation and apoptosis of HepG2 cells in vitro were investigated by EdU staining and TUNEL staining. The apoptosis was further confirmed by flow cytometry.The effect of HA-EPA nanoparticles on the migration and invasion of HepG2 cells was demonstrated by transwell and invasion experiments.The results of 1H NMR showed that HA-EPA was successfully synthesized, and the grafting rate of EPA on HA was (40 ± 5) %. The structure of HA-EPA was further confirmed by FT-IR.The particle size was (162.5 ± 10.2) nm, and the potential was -(4.47 ± 0.31) mV.MTT results showed that, with the prolongation of drug treatment time, HA-EPA showed a better inhibitory effect on the activity of HepG2 and Huh-7 cells than EPA under the same EPA content.After treated for 48 hours, the toxicity of HA-EPA to LX-2 cells was less than that of EPA.The results of 24-hour proliferation, apoptosis, migration and invasion of HepG2 showed that, the graft of hyaluronic acid improved the ability of EPA to inhibit proliferation, promote apoptosis, migration and invasion of HepG2 cells (P < 0.001), indicating that grafting of HA can significantly enhance the inhibitory effect of EPA on liver cancer with some role in reducing toxicity.
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出版历程
  • 收稿日期:  2021-06-24
  • 修回日期:  2022-01-27
  • 刊出日期:  2022-02-24

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