汉黄芩素通过调节肠道菌群抑制结直肠癌的药效机制

Mechanism of antitumor efficacy of Wogonin in suppressing colorectal cancer by regulating gut microbiota

目的:探讨汉黄芩素调节肠道菌群,抑制结直肠癌(CRC)小鼠肿瘤发生的药效机制。方法:分别建立APCmin+基因缺陷小鼠模型和AOM/DSS炎癌转化小鼠模型,并给予0.2%脱氧胆醇(DCA)饮用水进行造模,随机分为正常组、模型组、汉黄芩素高、低剂量组,给予相应药物干预,隔日观察小鼠状态。处死小鼠后取结直肠组织,HE染色观察组织病理形态学变化;免疫荧光染色法检测小鼠瘤组织F4/80、Mannose表达;TUNEL染色观察汉黄芩素对肿瘤细胞凋亡的影响;通过16S rRNA测序技术检测各组小鼠肠道菌群的变化,并进行生物信息学分析。结果:与对照组比较,DCA的干预显著促进了CRC的发生,而汉黄芩素可以显著减少结直肠癌肿瘤的体积(P<0.01),HE及TUNEL染色结果显示,DCA干预减少了肿瘤细胞凋亡,而汉黄芩素促使肿瘤细胞发生凋亡;免疫荧光染色结果显示,DCA干预增加了小鼠瘤组织内F4/80、Mannose的表达,而汉黄芩素给药组小鼠瘤组织内F4/80、Mannose表达显著减少。LEfSe分析提示,经DCA造模后,瘤胃球-UCG-014属、相对丰度下降,拟杆菌、鼠杆菌、相对丰度显著上升。相较于模型组,汉黄芩素高剂量干预后大肠埃氏菌属-志贺氏菌属、副拟杆菌属的丰度显著下调,伊雷杆菌属、瘤胃球-UCG-014属的丰度上调。KEGG富集结果表明,相关肠道菌落调控CRC与碳水化合物代谢、癌症通路、氨基酸代谢密切相关。结论:汉黄芩素能够通过调节肠道菌群抑制DCA介导的结直肠癌的发生。

Objective:To investigate the mechanism of Wogonin in regulating gut microbiota and suppressing colorectal cancer(CRC)tumor formation.Methods:APCimit gene-deficient mouse model and AOM/DSS infammatory cancer transformation mouse model were established,and 0.2%DCA was used for modeling.The mice were randomly divided into normal,model,and high/low-dose Wogonin groups and given the corresponding drugs.Mouse status was observed every other day.After euthanizing the mice,colorectal tissue was obtained to observe the histopathological changes with HE staining,detect F4/80 and Mannose expression in tumor tissue with immunofluorescence staining,and observe the effect of Wogonin on tumor cell apoptosis with TUNEL staining.Changes in gut microbiota of mice in each group were detected by 16S rRNA sequencing technology,and bioinformatics analysis was performed.Results:Compared to the control group,DCA intervention significantly promoted CRC development.However,Wogonin significantly reduced the volume of colorectal cancer tumors(P<0.01).HE and TUNEL staining results showed that DCA intervention reduced tumor cell apoptosis,while Wogonin induced tumor cell apoptosis.Immunofuorescence staining results showed that DCA intervention increased the expression of F4/80 and Mannose in mouse tumor tissue,while the expression of F4/80 and Mannose in the Wogonin treatment group was significantly reduced.LEfSe analysis suggested that after DCA modeling,the relative abundance of Ruminococcaceae_UCG-014 decreased,and Muribaculum significantly increased.Compared to the model group,the high-dose Wogonin intervention significantly downregulated Escherichia-Shigella and Parabacteroides and upregulated Ileibacterium and Ruminococcaceae_UCG-014.KEGG enrichment results indicated that the regulation of CRC-related gut microbiota was closely related to carbohydrate metabolism,cancer pathways,and amino acid metabolism.Conclusion:Wogonin can inhibit the occurrence of colorectal cancer mediated by DCA by regulating intestinal flora.