京尼平所致肝细胞毒性的物质基础研究  

Exploring the material basis of genipin-induced hepatotoxicity in vitro

作　　者：张帆[1] 吕东霞 董毓松 秦佳琪 王国旭 骆洋 饶志[1] 魏玉辉[1] ZHANG Fan;LV Dong-xia;DONG Yu-song;QIN Jia-qi;WANG Guo-xu;LUO Yang;RAO Zhi;WEI Yu-hui(The First Hospital of Lanzhou University,Lanzhou 730000,China;The 1^(st)School of Clinical Medicine,Lanzhou University,Lanzhou 730000,China;School of Pharmacy,Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一医院,甘肃兰州730000 [2]兰州大学第一临床医学院,甘肃兰州730000 [3]兰州大学药学院,甘肃兰州730000

出　　处：《药学学报》2023年第10期3032-3039,共8页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(82174067,82004080,81960646);兰州大学2022年国家级大学生创新创业推荐项目资助(202210730179)。

摘　　要：本文通过探究京尼平闭环原型、开环异构体及其与氨基酸结合而成的栀子蓝的HepaRG肝细胞毒性,阐明京尼平所致肝细胞毒性的物质基础。考察不同温度、pH值及氨基酸对京尼平结构互变及栀子蓝形成的影响,明确可区分京尼平闭环和开环异构体的条件,以及栀子蓝形成的条件并制备不同氨基酸类型的栀子蓝冷冻干燥粉末;CCK-8法测定京尼平原型、开环异构体以及不同类型栀子蓝的肝细胞毒性。结果显示,酸性条件可抑制京尼平由闭环向开环的转化;伯氨基和京尼平开环结构是生成栀子蓝的必要条件,且两者最可能形成二氢吡啶环型的栀子蓝;37与80℃合成的栀子蓝存在明显差异;pH=5的酸性条件与正常培养条件(pH=7.4)相比,京尼平原型浓度显著升高,但细胞存活率并未明显降低;栀子蓝的细胞毒性显著低于京尼平开环结构,甚至部分类型栀子蓝呈现促进细胞生长的作用。研究结果表明,京尼平开环结构可能是造成肝细胞毒性的重要物质基础,为今后京尼平肝毒性机制等相关研究奠定了基础并提供了线索。The current study explored the hepatotoxicity among closed-ring genipin,open-ring tautomer of genipin and gardenia blue that generated from genipin and amino acid reaction using HepaRG cells to identify the material basis of genipin-induced hepatotoxicity in vitro.The effects of temperature,pH value and different kinds of amino acids on the chemical structure tautomerism between closed-ring and open-ring tautomer of genipin and the production of gardenia blue were investigated firstly,which aimed to explicit the conditions that could distinguish the closed-ring genipin and its open-ring tautomer,and the conditions generating gardenia blue,which were applied to prepare different kinds of gardenia blue;the CCK-8 kit was employed to analyze the hepatotoxicity of closed-ring genipin,open-ring tautomer of genipin and gardenia blue.From the results,it was found that,the structure transformation from close-ring to open-ring of genipin could be inhibited under the condition with acid environment;being essential groups to generate gardenia blue,the primary amino group and the open-ring tautomer of genipin reacting to generate the dihydropyridine ring was probably the key structure of gardenia blue;the structure characteristics existed apparent distinction at the reactive temperature of 37℃ and 80℃;compared to the culture condition with pH=7.4,the concentration of genipin with close-ring in culture medium was significantly increased at pH=5,but the cell viability did not decreased;the cell toxicity of gardenia blue was apparently lower than open-ring tautomer of genipin,and even some kinds of gardenia blue showed growth promoting effect on HepaRG cells.Here,it was suggested potentially that open-ring tautomer of genipin be the important material basis to induce hepatotoxicity,which could provide a cue and lay a foundation for the elucidation of the underlying mechanism of genipin-induced hepatotoxicity.

关 键 词：京尼平 开环异构体 栀子蓝 肝毒性 物质基础 

分 类 号：R966[医药卫生—药理学]