基于传统用法和成分组合研究胡桃楸的抗肿瘤有效成分组及其作用机理

Juglans mandshurica, a folk medicine reputed for its anticancer effect, was traditionally decocted together with egg. Only eating the medicated egg was enough for mild case, while for severe case the decoction and the medicated egg were all needed. According to the results of our research, the medicated egg and the hydrolysable tannins showed inhibitory effects in vivo. This traditional usage and the results of our group indicated that the decoction of and the egg decocted with Juglans mandshurica were all effective in anticancer and implied that there were effective constituents in the medicated egg. . For oral administered drugs, only constituents absorbed by living body might exhibit pharmacological effects. . In this project the constituents absorbed by medicated egg and living body were analyzed by LC-MSn. The combination of those constituents is designed by uniform distribution. For different combinations, the in vitro activities are determined by high through-put screening method using cancer cell as an target. While, the in vivo activities of those combinations are conducted on hollow fiber tumor model. To research of the interaction of multi-constituents, the theory of Loewe additivity and Bliss independence is used. The concentration-response relationship is analyzed by response surface method established by Carter W.H. After establishing the concentration-response equation, the combination with the best response will be found and the anti-cancer mechanism of the best combination will be researched. This project would provide a new research pattern for the combination of multiple constituents, and provide useful data for the research and development of new anticancer drug based on Juglans mandshurica.

胡桃楸资源丰富，抗肿瘤作用显著，但其药效物质基础及作用机理尚不明确。本课题组根据胡桃楸与鸡蛋同煮的传统用法，发现：含药鸡蛋及其吸收的鞣质显示出较强的抗癌活性。提出“分析含药鸡蛋中吸收的成分可有目标地缩小有效成分研究的范围”。胡桃楸采用口服给药，研究其体内吸收和代谢的成分可排除生物利用度低的成分，也从另一角度缩小了有效成分研究的范围。将含药鸡蛋吸收与体内吸收的成分相结合既能缩小有效成分的研究范围，又能全面囊括几乎所有活性成分，使成分组合研究成为可能。因此本课题拟根据胡桃楸与鸡蛋同煮口服的传统用法，通过液质分析含药鸡蛋和体内吸收、代谢的成分，首次采用活性指导分组的研究思路对这些成分进行组合，运用Loewe和Bliss模型研究多成分间相互作用，寻找最佳抗肿瘤有效成分组、研究其作用机理。本课题将为胡桃楸的新药研究和质量评价奠定基础，为中药有效成分组合研究提供新的模式。

胡桃楸资源丰富，抗肿瘤作用显著，但其抗肿瘤有效成分及作用机理仍不明确。本项目基于胡桃楸与鸡蛋同煮的传统用法，通过分析含药鸡蛋中吸收的成分有目标地缩小有效成分研究的范围。同时，研究其体内吸收代谢的成分排除生物利用度低的成分。在此基础上，通过吸收成分的组合设计寻找胡桃楸的抗肿瘤作用物质基础，通过网络药理学研究探讨胡桃楸的作用机理。.通过本项目的研究分析鉴定了胡桃楸水煎液中93个成分，包括鞣质、萘烷衍生物、有机酸类、黄酮、木脂素、蒽醌、二芳基庚烷、氨基酸和含氮化合物。确定了含药鸡蛋主要以浓度依赖的方式吸收了44个胡桃楸水煎液中的成分，大部分成分吸收率为1%~9%。.小鼠灌胃给予胡桃楸水提物后，血浆、尿液、胆汁、心、肝、脾、肺、肾中共检测到37个原型化合物和30个代谢物，原型化合物包括鞣质、有机酸、萘衍生物、二芳基庚烷、黄酮和多糖；代谢物包括葡萄糖醛酸化物、硫酸酯化物、葡萄糖醛酸化硫酸酯化物和异构化物。.从胡桃楸茎枝中分离富集了5个含量较高的成分：逆没食子酸、没食子酸、1,6-二-O-没食子酰基-β-D-葡萄糖、1,2,6-三-O-没食子酰基-β-D-葡萄糖、1,2,3,6-四-O-没食子酰基-β-D-葡萄糖。前4个成分均能被小鼠体内吸收。.通过成分组合设计，发现上述胡桃楸中的主要成分组合具有较好的体外抗肿瘤活性；成分间相互作用主要为加和与拮抗作用，而非协同作用。.通过网络药理学研究从成分和靶点相互作用角度探讨了胡桃楸的抗胃癌作用靶点，MAPK（丝裂原活化蛋白激酶）1/3/8是药物与疾病互作的重要靶蛋白，MAPK家族可能为胡桃楸抗肿瘤作用的重要靶点。.上述研究内容已发表4篇核心期刊论文，1篇普通论文，2篇SCI收录论文；培养硕士研究生3名；获得辽宁省自然学术成果三等奖1项；发表3篇会议论文，2篇被评为优秀论文。.本项目研究丰富了胡桃楸化学成分、抗肿瘤作用研究的数据，从传统用法角度揭示了胡桃楸的抗肿瘤药效物质基础，探讨了其作用机理，为基于胡桃楸的抗肿瘤新药研究奠定了基础。