黄葵总酮对大鼠顺铂化疗减毒增效作用机理的代谢组学研究

Cisplatin is one of the most widely used and most potent chemotherapy drugs. However, side effects in normal tissues and organs, notably nephrotoxicity in the kidneys, limit the use of cisplatin and related platinum-based therapeutics. The underlying mechanism of this nephrotoxicity has not fully understood, and there still lack of the effective renoprotective approaches without compromising the anti-tumour activity of cisplatin. Our preliminary study demonstrated that the total flavonoids extracted from flowers of Abelmoschus manihot (named as HKZT) could effectively attenuate cisplatin-induced nephrotoxicity. Moreover, most major constituents contained in HKZT were proved to possess remarkable anti-tumour activities. Accordingly, we proposed that HKZT has the potential to be an ideal renoprotective preparation for cisplatin-induced nephrotoxicity. Metabonomic approaches has unique advantages in elucidating the complex multi-pathway mechanisms of cisplatin-induced nephrotoxicity, and characterizing the holistic pharmacological properties of HKZT displayed as multi-component, multi-target and multi-pathway. Considering this, in the current project, an RP/HILIC-UPLC-TOFMS metabonomic approach is employed to investigate the evolution process of cisplatin-induced nephrotoxicity, the renoprotective action of HKZT on the nephrotoxicity, as well as the effect of HKZT on the anti-tumour activity of cisplatin, in normal and tumor-bearing rats, thereby screening the metabolite-level biomarkers to elucidate the mechanism of nephrotoxicity, renoprotection and chemotherapy enhancement. This project would fully demonstrate the feasibility of using HKZT as an ideal renoprotection for cisplatin-based therapy, providing the scientific basis to support the application of HKZT-related prescriptions in clinical cancer therapy.

顺铂是当前肿瘤联合化疗的主流用药，但剂量限制性肾毒性，极大限制顺铂的临床应用。目前顺铂肾毒作用机理尚未完全明确，仍缺乏显著降低肾毒性且不影响化疗效果的保护剂。本课题组前期研究表明，中药黄蜀葵花提取物黄葵总酮（HKZT）可以有效缓解顺铂所致机体肾毒性；同时研究显示，金丝桃苷等黄葵中主要化学成分均具有明确的抗肿瘤活性。因此，本课题设想HKZT可作为理想的顺铂化疗保护剂。而代谢组学在阐明顺铂多重复杂致毒机理和HKZT多成分多靶点协同增效的整体药效方面具有独特优势。基于此，本研究拟从正常大鼠和荷瘤大鼠水平，采用基于RP/HILIC-UPLC-TOFMS的代谢组学研究手段，系统考察HKZT对顺铂肾毒性及其化疗疗效的影响，以期从代谢物组层面筛选顺铂致毒、HKZT减毒和增效相关生物标志物，揭示其内在作用机理。本课题可全面评价HKZT作为顺铂化疗肾保护剂的合理性，拓展黄葵制剂在肿瘤临床治疗中的应用。

顺铂是当前肿瘤联合化疗的主流用药，但剂量限制性肾毒性，极大限制顺铂的临床应用。前期研究表明，中药黄蜀葵花提取物黄葵总酮（HKZT）可以有效缓解顺铂所致机体肾毒性；同时研究显示，金丝桃苷等黄葵中主要化学成分均具有明确的抗肿瘤活性。因此，HKZT或许可作为一种理想的顺铂化疗保护剂。.基于此，本课题首先对HKZT提取物中的38个黄酮类成分及其在大鼠血液和尿液中的16个代谢产物进行定性分析，而后从正常大鼠和荷瘤小鼠两个层面，采用基于RP/HILIC-UPLC-TOFMS的代谢组学研究手段，从尿液、血清和肾脏组织代谢物组层面考察HKZT对顺铂肾毒性及其化疗疗效的影响。.结果发现：（1）HKZT可显著改善顺铂导致的正常大鼠体重降低，血清肌酐和尿素氮等生化指标的升高，以及尿液、血清和肾脏的代谢轮廓异常偏移，减毒机理与抑制氨基酸、核苷酸、甘油磷脂和寡肽（血清）类代谢，激活二肽（肾脏）、寡肽（肾脏）和饱和溶血磷脂胆碱类代谢，以及调节不饱和溶血磷脂胆碱和磷脂酰乙醇胺的代谢异常有关。.（2）HKZT可一定程度上改善顺铂所导致的荷瘤小鼠生化指标异常，如Crea和CysC水平异常升高等，其减毒作用与上调包括磷脂酰胆碱、鞘磷脂、二酯、饱和溶血磷脂酰胆碱、溶血磷脂、缩醛磷脂、甘油三酯和不饱和溶血磷脂酰胆碱在内的磷脂类化合物和肉碱，下调磷脂酰肌醇代谢，以及回调氨基酸代谢、核苷酸代谢和磷脂酰乙醇胺代谢紊乱等途径密切相关；.（3）单用HKZT可在一定程度上抑制荷瘤小鼠肿瘤生长，减少肝脾肿大和肾脏萎缩，降低荷瘤小鼠血清中ALT和LDH水平，升高UA水平，从而表现出一定的抗肿瘤作用。抗肿瘤作用与激活前列腺素、血清中磷脂酰胆碱、溶血磷脂、谷氨酸和花生四烯酸代谢，下调核苷酸、肾脏中磷脂酰胆碱、甘油磷脂和二肽类代谢，调节氨基酸和亚油酸代谢等途径密切相关。.该研究可为HKZT用作顺铂化疗保护剂提供初步的科学依据。