Integrated metabolomics and gut microbiota to reveal the anti-tumor mechanism of Jinfu'an decoction in tumor-bearing mice

Introduction: Jinfu’an Decoction (JFAD), a traditional Chinese medicine, is used to treat lung cancer and has shown significant anti-tumor effects in clinical and experimental studies. This study integrates metabolomics and gut microbiota analysis to elucidate JFAD’s anti-tumor mechanisms.Methods: A suspension of A549-luc cells, approximately 1×10^6 in number, was injected subcutaneously into the right axilla of mice to establish a tumor-bearing nude mouse model. Mice were randomly assigned to four groups: model group (MG), low-dose JFAD (JFAD-L), medium-dose JFAD (JFAD-M), and high-dose JFAD (JFAD-H), receiving treatments via gavage for 21 days. Additionally, three nude mice formed the normal group (NG), receiving no treatment. Changes in gut microbiota and serum metabolites were assessed using 16S rRNA gene sequencing and UHPLC-QE-MS non-targeted metabolomics.Results: JFAD may help restore the balance of intestinal flora in mice with lung cancer to a more normalized state. Our findings indicate that JFAD increases the abundance of Bacteroidia and decreases the presence of Firmicutes and Clostridia, thereby altering intestinal bacterial composition. Primary metabolic pathways associated with significant differences include nicotinate and nicotinamide metabolism, glycine, serine and threonine metabolism, and pyrimidine metabolism. A key differential metabolite identified was succinic acid, part of the central carbon metabolism pathway in cancer. Succinic acid showed a negative correlation with gut microbiota families Tannerellaceae and Campylobacterota. In the MG group, essential amino acid levels were markedly diminished but were significantly elevated after JFAD-M intervention. KEGG pathway analysis identified these amino acids as being linked to the PI3K/AKT and mTOR signaling pathways.Discussion: JFAD regulates the homeostasis of intestinal flora and influences amino acid and succinic acid metabolism through various pathways. These mechanisms could serve as potential targets for JFAD in inhibiting lung cancer invasion and metastasis.

引言：金复安煎剂（Jinfu’an Decoction, JFAD）是一种传统中药，用于肺癌治疗，且在临床与实验研究中均展现出显著的抗肿瘤效应。
本研究整合代谢组学与肠道菌群分析技术，以阐明金复安煎剂的抗肿瘤作用机制。
方法：将数量约为1×10^6的A549-luc细胞悬液接种于裸小鼠右侧腋下，构建荷瘤裸鼠模型。
将小鼠随机分为四组：模型组（MG）、低剂量金复安煎剂组（JFAD-L）、中剂量金复安煎剂组（JFAD-M）及高剂量金复安煎剂组（JFAD-H），各组均通过灌胃给药，持续21天；另设3只未接受任何处理的裸小鼠作为正常对照组（NG）。
采用16S rRNA基因测序与超高效液相色谱-四级杆-静电场轨道阱高分辨质谱（UHPLC-QE-MS）非靶向代谢组学技术，检测肠道菌群与血清代谢物的变化。
结果：金复安煎剂可帮助肺癌小鼠的肠道菌群平衡恢复至更接近正常的状态。
本研究结果显示，金复安煎剂可提高拟杆菌纲（Bacteroidia）的丰度，同时降低厚壁菌门（Firmicutes）与梭菌纲（Clostridia）的占比，进而改变肠道菌群组成。
存在显著差异的核心代谢通路包括烟酸与烟酰胺代谢、甘氨酸-丝氨酸-苏氨酸代谢以及嘧啶代谢。
本研究鉴定出的关键差异代谢物为琥珀酸，其参与癌症的中枢碳代谢通路。
琥珀酸与肠道菌群的坦纳菌科（Tannerellaceae）及弯曲菌门（Campylobacterota）呈负相关。
模型组小鼠的必需氨基酸水平显著降低，而中剂量金复安煎剂干预后该水平显著回升。
KEGG通路分析显示，这些必需氨基酸的代谢与PI3K/AKT及mTOR信号通路密切相关。
讨论：金复安煎剂可通过多种通路调控肠道菌群稳态，并影响氨基酸与琥珀酸代谢。
上述机制或可作为金复安煎剂抑制肺癌侵袭与转移的潜在作用靶点。