基于肠道菌群及代谢组学解析白花蛇舌草干预结肠炎-癌转化作用机制

From the perspective of intestinal flora，the mechanism of traditional Chinese intervention in the evolution of disease has becomes a hotspot in the field of traditional Chinese medicine and microbiology. Previous research has indicated that Hedyotis diffusa as one of the anticancer drugs can regulate cancerous intestinal flora. However, the mechanism of intestinal flora as the intervention target mediated carcinogenesis remains to be explained. Based on this,presumably, Hedyotis diffusa interposes canceration process by optimizing the intestinal flora structure and function, repairing endogenous metabolites, inhibition of inflammatory factors.The model mice of AOM/DSS induced colitis inflammation-cancer which given Hedyotis diffusa are used to be compared the structure of intestinal flora and its metabolites of dynamic change with the normal mice through the Illumina MiSeq high-throughput sequencing platform ,LC-HR-MS/MS ,and metabonomics method during the different stages of sequence evolution "inflammation-cancer". The cytokine antibody array technology was used to monitor the secretory expression of inflammatory cytokines in mice and to construct the interaction network changes in secretion. This research investigated the changes of intestinal microflora, characteristic metabolites and inflammatory factors in the process of carcinogenesis, inorder to reveal the mechanisms for canceration intervened by intestinal flora based on the regulation of Hedyotis diffusa,to provide a new way for the research of the innovation pattern of "intestinal flora +" traditional Chinese medicine.

从肠道菌群角度阐释中药干预疾病演变的作用机制是目前中医药、微生物领域共同关注的研究热点。前期研究表明，抗癌中药白花蛇舌草可调节癌变机体肠道菌群，但肠道菌群作为干预“靶点”介导癌变的作用机制尚待阐释。基于此，推测白花蛇舌草通过优化肠道菌群结构和功能，修复内源性代谢物轮廓、抑制炎症因子干预机体癌变进程。拟采用AOM/DSS诱导的结肠“炎-癌”小鼠模型，给药白花蛇舌草，并在“炎-癌”不同序列演进阶段，采用Illumina MiSeq高通量测序平台结合LC-HR-MS/MS代谢组学方法，比较正常及结肠癌小鼠肠道菌群结构及相应代谢产物的动态变化差异；采用炎症因子抗体芯片等技术，监测小鼠炎症因子分泌表达，构建炎症因子分泌变化交互作用网络图，探讨癌变过程中与此相关性肠道菌群、特征性代谢产物及炎症因子的变化，揭示白花蛇舌草调节肠道菌群干预癌变机制,为“肠道菌群+”中药创新性研究模式提供新思路。

从肠道菌群角度阐释中药干预肿瘤等疾病演变的作用机制是当前中医药和微生物领域十分关注的研究热点。本研究团队前期研究表明，中药白花蛇舌草可调节癌变机体肠道菌群，但以肠道菌群作为干预“靶点”介导癌变的作用机制尚待阐释。基于此，推测白花蛇舌草可通过优化肠道菌群结构和功能，修复内源性代谢物轮廓、抑制炎症因子干预机体癌变进程。项目组首先构建了AOM/DSS诱导结肠炎-癌小鼠模型。主要通过模拟溃疡性结肠炎的癌变过程中肠道黏膜上皮持续的损伤与修复病程，利用化学致癌剂AOM与致炎剂DSS的协同作用，成功构建与人类UC-CRC相似特征的“炎症-不典型增生-癌症”小鼠模型。进一步开展不同序列演变阶段肠道菌群谱研究。主要以构建的AOM/DSS诱导结肠炎-癌小鼠模型为对照，在小鼠模型演变阶段中采取粪便，采用Illumina MiSeq高通量测序平台检测肠道菌群动态变化，分析结肠炎癌转化过程中肠道菌群谱的动态变化，并于正常小鼠比较，探讨肠道菌群组成差异。继续开展白花蛇舌草给药后代谢产物研究。并在不同阶段取血、粪便等生物样本，采用代谢组学方法，基于LC-HR-MS/MS技术检测正常小鼠、模型小鼠及给药小鼠生物样本中代谢产物，并根据代谢谱差异，探讨结肠炎-癌转化的标志性代谢物及白花蛇舌草对代谢物的影响。以AOM/DSS小鼠模型为研究对象，采用炎症因子抗体芯片技术，监测结肠炎-癌小鼠不同演变阶段小鼠炎症因子分泌表达，探讨白花蛇舌草干预炎-癌转化过程对分泌因子变化的影响。研究团队通过本项目的实施，力求揭示白花蛇舌草调节肠道菌群干预癌变机制,为“肠道菌群+”中药创新性研究模式提供新研究模式。