不同中医证候的结肠癌肠道菌群特点及肝气郁结型结肠癌肠道菌群的促瘤作用

Characteristics of gut microbiota of colon cancer in different traditional Chinese medicine syndromes and the tumor-promoting effect of gut microbiota of liver qi stagnation type colon cancer

目的:探究不同中医证候的结肠癌肠道菌群特点及肝气郁结型结肠癌肠道菌群的促瘤作用。方法:对入组48例患者进行中医辨证,收集患者粪便,通过16S rRNA测序明确菌属多样性与特异性,明确肝气郁结型结肠癌患者肠道菌群与证候量表相关性。通过粪菌移植技术,初步探索肝气郁结型结肠癌促瘤作用机制。构建氧化偶氮甲烷/葡聚糖硫酸钠(AOM/DSS)模型,观察肿瘤增殖情况。结果:各中医证候间肠道菌群组成存在差异。其中,肝气郁结型特异性及多样性较高。肝气郁结型结肠癌患者中Prevotella、Escherichia、Bacteroides、Granulicatella等菌属与证候量表中嗳气、胁肋胀痛、呕逆以及脉弦等存在密切联系。经肝气郁结型结肠癌患者粪菌移植后,小鼠第6、12周体质量下降(P<0.01),生存时间缩短(P<0.05),肠黏膜发生了高级别上皮内瘤变。血清中IL-2、IL-4、IL-10、IL-13、TGF-β及TNF-α炎症因子水平上升(P<0.01)。AOM/DSS模型中,肝气郁结菌液干预小鼠肿瘤数量显著增加(P<0.05),总生存时间减少(P<0.05),Ki-67表达明显增加。结论:不同中医证候的结直肠癌患肠道菌群有明显差异。肝气郁结型结肠癌患者肠道菌群失调后,通过炎症水平增高、肿瘤细胞增殖,促进了结肠癌的发生发展。

Objective:To investigate the characteristics of intestinal flora of colon cancer patients with different traditional Chinese medicine(TCM)symptoms and the tumor-promoting effect of intestinal flora of liver qi stagnation type.Methods:A total of 48 patients were identified by TCM,and the genetic diversity and specificity of the feces were determined by 16S rRNA sequencing,and the correlation between the intestinal flora and the evidence scale of colon cancer patients with liver qi stagnation.The tumor-promoting mechanism of liver qi stagnation colon cancer was initially explored by fecal transplantation.AOM/DSS model was constructed to observe tumor proliferation.Results:There were differences in the composition of intestinal flora among the TCM evidence.Among them,the specificity and diversity were higher in the liver qi stagnation type.Prevotella,Escherichia,Bacteroides,and Granulicatella were closely associated with belching,painful distention of the ribs,vomiting,and chordal pulse in the syndrome scale.After transplantation of fecal bacteria in patients with hepatic colon cancer,the body mass of mice decreased at the 6th and 12th week(P<0.01),the survival time was shortened(P<0.05),and high-grade intraepithelial neoplasia occurred in the intestinal mucosa.Serum levels of inflammatory factors such as IL-2,IL-4,IL-10,IL-13,TGF-βand TNF-αincreased(P<0.01).In the AOM/DSS model,the number of tumors increased(P<0.05),the overall survival time decreased(P<0.05)and Ki-67 expression increased in mice with liver qi fungus solution intervention.Conclusion:There are significant differences in the intestinal flora of colon cancer patients with different TCM syndromes.The dysregulation of intestinal flora in colon cancer patients with liver qi stagnation type promot the development of colon cancer through the increase of inflammation level and proliferation of tumor cells.