健脾解毒方（JPJDF）是一种传统中药汤剂，因其抗癌特性而在临床上使用，特别是在结直肠癌（CRC）中。本研究旨在探讨健脾解毒方对结直肠癌的治疗作用并阐明其潜在的分子机制。重点关注其对体外和体内缺氧诱导因子 1 α (HIF1α) 和癌症相关成纤维细胞 (CAF) 的影响。使用 UPLC-Q-TOF-MS 鉴定 JPJDF 的成分。建立化学诱导结直肠癌模型并用JPJDF治疗以评估其效果。测量肿瘤大小，并进行组织病理学分析，以检查 JPJDF 对 CRC 的调节潜力。通过网络药理学、分子对接和转录组学预测了JPJDF的作用机制。使用涉及 CRC 细胞和 CCD-18Co 成纤维细胞的共培养技术来评估 JPJDF 对成纤维细胞活化的影响。使用 CCK-8 增殖、克隆形成和凋亡测定评估 HIF1α 对 CAF 的影响，并通过 qPCR 和蛋白质印迹定量差异标记表达。药效学评估表明，JPJDF 在不影响体重的情况下缩小了肿瘤大小，表明其在临床中的安全性。化学诱导的小鼠CRC模型。网络药理学分析结合分子对接和转录组学，揭示 JPJDF 调节 HIF-1 信号通路，并确定 HIF1α 是 JPJDF 抗 CRC 作用的潜在靶点。 JPJDF 通过抑制 HIF1α 减弱成纤维细胞活化、减少 α-SMA 表达和 POSTN 分泌，有效抑制体内 CRC 生长。 CRC 细胞中 HIF1α 的敲低抑制了成纤维细胞增殖和克隆形成，而过度表达则促进了这些过程。此外，下调 HIF1α 抑制成纤维细胞中的 α-SMA 和 POSTN 表达，而过度表达则增强成纤维细胞激活。JPJDF 成为通过靶向 HIF1α 抑制 CAF 激活的有前途的治疗候选药物，为在 CRC 治疗中调节成纤维细胞对 CAF 的激活提供潜在途径。版权所有 © 2024 年。由 Elsevier B.V 出版。

Jianpi Jiedu Formula (JPJDF) is a traditional Chinese medicinal decoction clinically used for its anti-cancer properties, particularly in colorectal cancer (CRC).This study aims to investigate the therapeutic effects of JPJDF on CRC and elucidate its potential molecular mechanisms, with a focus on its impact on hypoxia-inducible factor 1 alpha (HIF1α) and cancer-associated fibroblasts (CAFs) both in vitro and in vivo.UPLC-Q-TOF-MS was used to identify the constituents of JPJDF. A chemical-induced colorectal cancer model was established and treated with JPJDF to evaluate its effects. Tumor size was measured, and histopathological analyses were performed to examine JPJDF's regulatory potential on CRC. The functional mechanism of JPJDF was predicted through network pharmacology, molecular docking, and transcriptomics. Co-culture techniques involving CRC cells and CCD-18Co fibroblasts were used to assess JPJDF's impact on fibroblast activation. The effects of HIF1α on CAFs were evaluated using CCK-8 proliferation, clonal formation, and apoptotic assays, with differential marker expression quantified via qPCR and Western blotting.Pharmacodynamic assessment demonstrated that JPJDF reduced tumor size without affecting body weight, indicating its safety in the chemical-induced murine CRC model. Network pharmacology analysis, combined with molecular docking and transcriptomics, revealed that JPJDF regulates HIF-1 signaling pathways and identified HIF1α as a potential target for JPJDF's anti-CRC effect. JPJDF effectively suppressed CRC growth in vivo by attenuating fibroblast activation, reducing α-SMA expression and POSTN secretion through HIF1α inhibition. HIF1α knockdown in CRC cells inhibited fibroblast proliferation and clonal formation, while overexpression promoted these processes. Additionally, downregulating HIF1α suppressed α-SMA and POSTN expression in fibroblasts, whereas overexpression enhanced fibroblast activation.JPJDF emerges as a promising therapeutic candidate for inhibiting CAFs activation by targeting HIF1α, offering potential avenues for modulating fibroblast activation towards CAFs in CRC therapy.Copyright © 2024. Published by Elsevier B.V.