背景:中医,健脾解毒汤(JPJDF),已经在结肠直肠癌(CRC)治疗中使用了40多年。JPJDF通过调节肠道微生物群及其代谢物来抑制CRC的潜力仍不确定。
目的:本研究旨在进一步探讨JPJDF在CRC中的治疗机制。
方法:使用偶氮甲烷(AOM)和葡聚糖硫酸钠(DSS)开发了CAC小鼠模型。肠组织和内容物进行16SrRNA基因测序和非靶向代谢组学分析。采用ELISA法测定血清IL-1β和TNF-α水平。免疫组织化学用于评估Ki67,ZO-1,Occludin,CD68和CD206。此外,进行蛋白质印迹以评估AhR和NF-κB的蛋白表达。
结果:JPJDF抑制AOM/DSS治疗小鼠的结直肠肿瘤发生,同时抑制肿瘤细胞增殖和上调紧密连接蛋白的表达。16SrRNA基因测序分析结果表明,JPJDF通过增加有益细菌的丰度来改变肠道微生物群的组成。此外,JPJDF减少色氨酸代谢物,能有效缓解CAC小鼠的炎症反应,显著恢复肠屏障功能。分子生物学实验证实,JPJDF抑制AhR和M2型肿瘤相关巨噬细胞的表达水平,从而促进抗肿瘤免疫并对CAC生长发挥抑制作用。
结论:JPJDF可以通过调节肠道菌群来调节色氨酸代谢-AhR通路,减少肠道炎症,改善肠道屏障功能,增强抗肿瘤免疫力,有效抑制CAC生长。
BACKGROUND: Traditional Chinese medicine, JianpiJiedu decoction (JPJDF), has been utilized in colorectal cancer (CRC) treatment for over forty years. The potential of JPJDF to inhibit CRC through modulation of intestinal microbiota and their metabolites remains uncertain.
OBJECTIVE: This study aims to further investigate the therapeutic mechanisms of JPJDF in CRC.
METHODS: CAC mouse models were developed using azoxymethane (AOM) and dextran sulfate sodium (DSS). Intestinal tissues and contents underwent 16S rRNA gene sequencing and untargeted metabolomics analysis. Serum levels of IL-1β and TNF-α were measured using ELISA. Immunohistochemistry was utilized to assess the expression of Ki67, ZO-1, Occludin, CD68, and CD206. Furthermore, western blotting was performed to evaluate the protein expression of AhR and NF-κB.
RESULTS: JPJDF inhibited colorectal tumourigenesis in AOM/DSS treated mice, while also suppressing tumor cell proliferation and upregulating the expression of tight junction proteins. The results of 16S rRNA gene sequencing analysis revealed that JPJDF altered intestinal microbiota composition by increasing the abundance of beneficial bacteria. Additionally, JPJDF reduced tryptophan metabolites, effectively alleviating inflammation and significantly restoring intestinal barrier function in CAC mice. Molecular biology experiments confirmed that JPJDF suppressed the expression levels of AhR and M2-type tumor-associated macrophages, thereby promoting anti-tumor immunity and exerting inhibitory effects on CAC growth.
CONCLUSIONS: JPJDF can regulate the tryptophan metabolism-AhR pathway by modulating the gut microbiota, reducing intestinal inflammation, improving intestinal barrier function, enhancing anti-tumor immunity, and effectively inhibiting CAC growth.