Zheng Zongmei, Du Yizhao, Jiang Hua, Shi Zhenpeng, Zhou Hailun, Jiao Lijing, Liu Peifeng, Gong Yabin
Department of Oncology, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 200437, China.
State Key Laboratory of Systems Medicine for Cancer, Shanghai Cancer Institute, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200032, China.
Phytomedicine. 2025 Jul;142:156766. doi: 10.1016/j.phymed.2025.156766. Epub 2025 Apr 12.
Colorectal cancer liver metastasis (CRLM) represents one of the most severe complications of colorectal cancer (CRC), often associated with unfavorable prognosis. The herbal formulation Kang-Ai-Jing-Fang (KAJF) has been employed clinically against CRC for several decades, although its precise mechanism of action remains elusive.
This study investigates the anti-metastatic potential of KAJF in CRLM, focusing on its modulatory effects on gut microbiota and inhibition of M2-like macrophage activation.
KAJF was administered orally to CRLM model mice established through intrasplenic injection of murine CRC cells. To elucidate the role of gut microbiota reshaped by KAJF, fecal microbiota transplantation (FMT) was utilized to assess its impact on CRLM inhibition. Core targets and active compounds were identified through network pharmacology and molecular docking. To characterize microbiota composition, metabolite profiles, and gene expression variations, 16S rRNA sequencing, untargeted liquid chromatography-mass spectrometry (LC-MS/MS) metabolomics, and transcriptomics analyses were performed.
KAJF demonstrated significant inhibitory effects on CRLM and ameliorated gut microbiota dysbiosis by enhancing the abundance of butyrate-producing bacteria (Lactobacillus, Bacteroides, Bifidobacterium). The therapeutic efficacy of KAJF-induced bacterial alterations in delaying CRLM and promoting butyrate-producing microbiota enrichment was further substantiated by FMT. Network pharmacology identified active ingredients in KAJF, including asperulosidic acid, polyphyllin II, ganoderenic acid B, calycosin, and ganoderic acid C, which exhibit substantial interactions with TLR4, PPARγ, SIRT1, PTGS2, and TNF. Molecular dynamics simulations and surface plasmon resonance (SPR) analysis demonstrated that ganoderic acid C2 exhibits a strong binding affinity for PPARγ. Moreover, KAJF administration led to a marked reduction in F4/80 CD206 macrophages and their associated cytokines (CCL17, CCL22, IL10, IL4, TGF), accompanied by a decrease in CD4 T cells and myeloid-derived suppressor cells (MDSCs), while increasing CD8 T cell populations.
This study demonstrates that KAJF mitigates CRLM, primarily through the regulation of gut microbiota and microbial metabolites, alongside inhibition of M2-like macrophage polarization. By integrating metabolomics, transcriptomics, and network pharmacology, this research elucidates the molecular mechanisms underpinning KAJF's therapeutic effects against CRLM, offering a promising approach for clinical intervention.
结直肠癌肝转移（CRLM）是结直肠癌（CRC）最严重的并发症之一，常与不良预后相关。抗癌经方（KAJF）作为一种中药制剂，已在临床上用于治疗CRC数十年，但其确切作用机制仍不清楚。
本研究探讨KAJF在CRLM中的抗转移潜力，重点关注其对肠道微生物群的调节作用和对M2样巨噬细胞激活的抑制作用。
通过脾内注射小鼠CRC细胞建立CRLM模型小鼠，口服给予KAJF。为了阐明KAJF重塑的肠道微生物群的作用，采用粪便微生物群移植（FMT）来评估其对CRLM抑制的影响。通过网络药理学和分子对接鉴定核心靶点和活性化合物。通过16S rRNA测序、非靶向液相色谱-质谱联用（LC-MS/MS）代谢组学和转录组学分析来表征微生物群组成、代谢物谱和基因表达变化。
KAJF对CRLM具有显著的抑制作用，并通过增加产丁酸菌（乳酸杆菌、拟杆菌、双歧杆菌）的丰度改善肠道微生物群失调。FMT进一步证实了KAJF诱导的细菌改变在延缓CRLM和促进产丁酸微生物群富集方面的治疗效果。网络药理学鉴定了KAJF中的活性成分，包括车叶草苷酸、重楼皂苷II、灵芝烯酸B、毛蕊异黄酮和灵芝酸C，它们与TLR4、PPARγ、SIRT1、PTGS2和TNF有大量相互作用。分子动力学模拟和表面等离子体共振（SPR）分析表明，灵芝酸C2对PPARγ具有很强的结合亲和力。此外，给予KAJF导致F4/80 CD206巨噬细胞及其相关细胞因子（CCL17、CCL22、IL10、IL4、TGF）显著减少，同时CD4 T细胞和髓源性抑制细胞（MDSC）减少，而CD8 T细胞群体增加。
本研究表明，KAJF主要通过调节肠道微生物群和微生物代谢物以及抑制M2样巨噬细胞极化来减轻CRLM。通过整合代谢组学、转录组学和网络药理学，本研究阐明了KAJF对CRLM治疗作用的分子机制，为临床干预提供了一种有前景的方法。
