基于多组分多靶点的琼玉膏降低顺铂肾毒性药效物质基础研究

Cisplatin is a major antineoplastic drug for the first-line treatment of solid tumors. However, nephrotoxicity is a frequent adverse effect of cisplatin, which restricts the use of high doses to maximize the therapeutic efficacy of cisplatin. In our previous work, we found that the ancient Chinese formula Qiong-Yu-Gao (QYG) could protect against cisplatin-induced nephrotoxicity without reducing the anti-tumor activity. QYG could reduce the renal accumulation of cisplatin, inhibit inflammatory responses and alleviate apoptosis in the pathological process. But the effective components of QYG still need further study. In recent decades, it has been widely acknowledged that Chinese herbal medicines apply a multi-component and multi-target approach in the treatment of diseases. Therefore, we raise the hypothesis that the bioactive compounds contained in QYG can be classified into several chemical groups; each group of chemical compounds characterized with similar pharmacophores may possess similar pharmacological activities and contribute to the efficacy of the whole formula. In this project, chemical groups of QYG will be separated and prepared, and each group of chemicals will be comprehensively characterized by a variety of modern analytical technique. Their pharmacological activities will be investigated through UPLC-QTOF-MS/MS-based metabolomic analysis and traditional pharmacological approaches using ICP-MS, qRT-PCR, ELISA, and western blot methods. The aim of this project is to reveal the effective components of QYG and how they contribute to the efficacy of QYG, which may provide scientific basis for the clinical application of QYG and the innovative drug research based on QYG.

顺铂为治疗多种实体瘤的一线用药，但肾毒性是限制其临床应用的主要因素。本课题组前期研究发现琼玉膏可通过减少顺铂肾积累、抑制炎症反应和抗凋亡等作用降低顺铂肾毒性，但其药效物质基础有待深入研究。基于中药多组分、多靶点作用特点，我们提出以下假说：琼玉膏含有多种药效组分，各组分具有的不同药理活性对琼玉膏降低顺铂肾毒性产生药效贡献。本项目拟运用多维色谱联用技术分离鉴定琼玉膏中环烯醚萜苷、苯乙醇苷、糠醛衍生物、皂苷、三萜酸、多糖等组分；利用小鼠顺铂肾损伤模型，采用基于UPLC-QTOF-MS/MS的代谢组学方法、ICP-MS、qRT-PCR、ELISA和Western blot等多学科交叉技术，综合评价各组分在抑制顺铂肾积累、炎症反应和细胞凋亡等方面的药理活性，阐明各组分对琼玉膏降低顺铂肾毒性的药效贡献，揭示琼玉膏的药效物质基础及多组分作用机制，为琼玉膏的临床应用和基于琼玉膏的创新药物研究提供科学依据。

顺铂为治疗多种实体瘤的一线用药，但肾毒性是限制其临床应用的主要因素。本课题组前期研究发现琼玉膏可通过减少顺铂肾积累、抑制炎症反应等作用降低顺铂肾毒性，但其药效物质基础有待深入研究。本项目以“组分—药效”关系为切入点，运用UPLC-QTOF-MS/MS、HPGPC-ELSD等技术制备分析琼玉膏中地黄醇溶性成分（环烯醚萜苷、苯乙醇苷和糠醛衍生物）、人参皂苷、茯苓三萜酸和多糖等组分；利用小鼠顺铂肾损伤模型，采用ELISA、ICP-MS、qRT-PCR、免疫组化、代谢组学等检测方法，综合评价各组分在改善肾功能、抑制炎症反应、抑制顺铂累积、调节免疫功能等方面的药理活性，阐明各组分对琼玉膏降低顺铂肾毒性的药效贡献。实验结果显示，地黄醇溶性成分、人参皂苷和琼玉膏多糖均可缓解顺铂引起的体重下降。地黄醇溶性成分和琼玉膏多糖可抑制顺铂引起的胸腺指数下降，人参皂苷可抑制顺铂引起的脾脏指数下降。肾功能测试中，地黄醇溶性成分、人参皂苷及琼玉膏多糖均可降低血清肌酐和尿素氮浓度水平，地黄醇溶性成分可提高排尿量。地黄醇溶性成分、人参皂苷及琼玉膏多糖可通过抑制Ctr1、Oct2表达和提高MATE1表达降低顺铂在肾脏的积累。地黄醇溶性成分、人参皂苷及琼玉膏多糖可降低外周血IL-6水平，琼玉膏多糖可上调外周血IL-10水平。从肠道免疫调节角度探讨了茯苓不同化学组分抗顺铂肾损伤的保护作用。结果显示茯苓水溶性多糖可缓解顺铂引起的结肠形态结构异常和炎症反应。血清代谢组学研究结果表明，茯苓水溶性多糖对顺铂造成的代谢紊乱具有较好的回调效果，调节作用主要涉及嘌呤代谢、硫胺素代谢和不饱和脂肪酸的生物合成等代谢通路。本项目初步揭示了琼玉膏的药效物质基础及多组分作用机制，可为琼玉膏的临床应用和基于琼玉膏的创新药物研究提供科学依据。