基于脑-肠轴色氨酸代谢通路靶向分析的黄芩汤防治伊立替康胃肠道毒性的作用机制研究

Irinotecan is a chemotherapy usually used in clinic. But it's gastrointestinal toxicity affects the cancer patient's quality of life and the rate of chemotherapy completion. Huang-Qin decoction was proved to be a modulator of chemotherapeutic agent in cancer therapy. Gut-brain axis and tryptophan pathway both play key roles in gastrointestinal disease. The aim of this study is to dig deep into the metabolic regulation of Huang-Qin Decoction on preventing the gastrointestinal toxicity induced by Irinotecan. To achieve this, a gut-brain axis tryptophan pathway targeted analysis method is developed using LC/MS/MS coupled with precolumn derivertization first, and then the change trends of tryptophan pathway related metabolites were compared between three groups (control, delayed-onset diarrhea caused by Irinotecan, and Huang-qin Decoction) and four type biological samples (hypothalamus, stomach, small intestine, and serum). The predictable success of this project would add value to elucidate the molecular mechanism of Huang-Qin Decoction on preventing the gastrointestinal toxicity induced by Irinotecan, verify the feasibility of using tryptophan pathway as target for early diagnosis and treatment for chemotherapy induced gastrointestinal toxicity, and set up an example in the field of using TCMs as adjuvant treatment during chemotherapy for cancer.

伊立替康是结直肠癌一线化疗药物，但其严重的胃肠道毒性限制了临床应用。中医治疗下痢的经方黄芩汤被证实能有效防治化疗所致的胃肠道毒性。脑肠互动和色氨酸代谢通路异常被证实与胃肠道疾病发生、发展有密切关联。在前期研究发现的基础上，为深入揭示黄芩汤防治伊立替康胃肠道毒性的作用机制，本项目以脑-肠轴色氨酸代谢通路为切入点，以伊立替康致大鼠迟发性腹泻模型为研究载体，针对低丰度、弱响应目标化合物结构特点，建立质谱导向性的基于稳定同位素衍生化液质联用技术的脑-肠轴四个部位（下丘脑、胃窦、小肠和血清）色氨酸代谢通路靶向分析方法，通过组间横向比较和生物样本间纵向比较，筛选参与脑肠互动调节的结点生物标记物，探索从脑-肠轴色氨酸代谢通路靶向代谢调节角度揭示黄芩汤防治伊立替康胃肠道毒性的分子机制，验证该通路作为化疗药物胃肠道毒性早期诊断及药物治疗网络靶标的可行性，也为中西药联用治疗恶性肿瘤的研究提供新的思路和借鉴。

伊立替康是结肠直肠癌一线化疗药物，但其严重的胃肠道毒性限制了临床应用。中医治疗下痢的经方黄芩汤被证实能有效防治伊立替康化疗所致的胃肠道毒性。本项目以伊立替康致大鼠迟发性腹泻模型为研究载体，以血清、脑、肠等相关样品为分析对象，通过基于LC/MS和GC/MS的非目标代谢组学物质筛选，发现伊立替康胃肠道毒性发生发展与氨基酸、能量和脂质代谢相关，主要涉及色氨酸、苯丙氨酸和酪氨酸合成代谢通路、胆酸合成代谢通路以及甘氨酸、丝氨酸和苏氨酸代谢通路，黄芩汤干预可回调氨基酸代谢中苯丙氨酸、酪氨酸和色氨酸合成代谢的紊乱。为进一步目标性地从脑-肠轴色氨酸代谢通路靶向代谢调节角度揭示黄芩汤防治伊立替康胃肠道毒性的分子机制，本项目首次建立了质谱导向性的基于配对衍生化液质联用技术的脑-肠轴四个部位（下丘脑、胃、小肠和血清）色氨酸代谢通路靶向分析方法，解决了生物基质中内源性小分子物质丰度低、响应弱的问题，可用于色氨酸代谢通路中12种代谢物的准确定量分析，为后续深入揭示黄芩汤防治伊立替康胃肠道毒性的作用机制、寻找化疗药物胃肠道毒性早期诊断及药物治疗网络靶标奠定了基础。本项目相关研究目前已发表论文10篇，其中SCI论文9篇。