芥菜提取物调节结肠癌小鼠肠道菌群及信号通路Wnt/JNK和NF-κB p65/COX-2

Mustard Leaf Extract Regulates Gut Microbiota and Wnt/JNK and NF-κB p65/COX-2 Signaling Pathways in a Mouse Colon Cancer Model

摘要:

探讨芥菜硫代葡萄糖苷（Glucosinolate, GSL）提取物对结肠癌的恢复作用。紫外法检测芥菜总GSL，氧化偶氮甲烷联合葡聚糖硫酸钠诱导Balb/c小鼠建立结肠炎性相关的结肠癌模型，小鼠分为对照组、模型组和芥菜GSL提取物低、高剂量组，样品干预9周后，分析炎症因子水平以及NF-κB p65/COX-2和Wnt/JNK信号通路的表达，通过16S rRNA高通量测序分析肠道菌群。结果显示，芥菜总GSL含量高达127.11 μmol/g。与模型组比较，芥菜GSL提取物高剂量组小鼠体质量和结肠长度分别增加了42.11%和20.37%（P<0.05），结肠肿瘤个数显著减少（P<0.05），结肠组织中IL-6、TNF-α和IL-17表达水平分别降低了46.88%、70.22%、20.43%，IFN-γ表达水平升高了98.55%，NF-κB p65、COX-2、Wnt和JNK的蛋白表达分别降低了60.91%、15.57%、63.31%、96.58%（P<0.05）；芥菜GSL提取物组显著回调了Bacteroides、Turicibacter、Ruminococcaceae UCG-014、Ruminiclostridium_5、Akkermansia、norank f Clostridiales vadinBB60 group、Prevotellaceae UCG-001 和Blautia等菌的丰度，恢复了肠道微生物的平衡状态。综上，芥菜GSL提取物通过改善结肠癌小鼠病理状态、调节炎症因子、信号通路和肠道微生物，展现辅助治疗结肠癌的潜力。

Abstract:

The therapeutic effects of mustard leaf extract, glucosinolate (GSL), on colon cancer were investigated. Total GSL content in mustard leaf was quantified using UV spectrophotometry. A colon cancer model associated with colitis was developed in Balb/c mice using azoxymethane and dextran sulfate sodium. The mice were divided into control, model, and low/high-dose mustard leaf GSL groups. After a 9-week trial, inflammatory cytokine levels and NF-κB p65/COX-2 and Wnt/JNK signaling pathway activities were analyzed. The gut microbiota was examined through high-throughput sequencing of 16S rRNA. Total GSL content in mustard leaf reached 127.11 μmol/g. Compared with the model group, the high-dose mustard leaf GSL group showed significant respective increases in weight and colon length (42.11% and 20.37%, respectively (P<0.05)), a significant reduction in the number of colon tumors (P<0.05), decreases in IL-6, TNF-α, and IL-17 expression in colon tissue by 46.88%, 70.22%, and 20.43% respectively, a 98.55% increase in IFN-γ expression, and significant decreases in NF-κB p65, COX-2, Wnt, and JNK protein expression by 60.91%, 15.57%, 63.31%, and 96.58%, respectively (P<0.05). The abundances of Bacteroides, Turicibacter, Ruminococcaceae UCG-014, Ruminiclostridium_5, Akkermansia, norank f Clostridiales vadinBB60 group, Prevotellaceae UCG-001, and Blautia were significantly altered by mustard leaf GSL, resulting in a restoration of a healthy microbial balance in the gut. In summary, leaf mustard GSL is a potential adjuvant therapy for colon cancer, as evidenced by its capacity to alleviate pathological conditions and modulate inflammatory cytokines, signaling pathways, and gut microbiota in mouse models of colon cancer.