基于高通量测序技术研究芍药汤对溃疡性结肠炎大鼠肠道菌群的影响

目的观察芍药汤对TNBS诱导的溃疡性结肠炎大鼠肠道菌群的影响,从菌群紊乱角度探讨其治疗溃疡性结肠炎的作用机制。方法将50只SD大鼠随机分为正常组、模型组、抗生素组、芍药汤组和美沙拉嗪组,每组10只。采用5%TNBS+50%乙醇联合诱导溃疡性结肠炎模型,第8日开始每日对应给予生理盐水、抗生素、芍药汤、美沙拉嗪灌胃7 d,HE染色观察结肠组织病理学变化,ELISA检测血清炎症因子含量,运用高通量测序技术检测大鼠粪便肠道菌群的变化。结果与正常组比较,模型组大鼠结肠损伤严重,血清白细胞介素(IL)-6、IL-1β、肿瘤坏死因子-α(TNF-α)含量显著升高(P<0.01);与模型组比较,芍药汤组、美沙拉嗪组、抗生素组大鼠结肠损伤减轻,血清IL-6、IL-1β、TNF-α含量显著下降(P<0.01,P<0.05);模型组大鼠菌群丰度、多样性较正常组均降低,芍药汤组及美沙拉嗪组肠道菌群丰度及多样性均升高;与模型组比较,芍药汤组及美沙拉嗪组门分类的厚壁菌门、变形菌门和属分类的梭菌属、颤螺旋菌属、乳酸菌属、普雷沃菌属、瘤胃球菌属、帕拉普氏菌属、罗斯氏菌属丰度均增加,门分类的拟杆菌门和属分类的拟杆菌属丰度均减少。结论肠道菌群与体内细胞因子分泌和表达密切相关,芍药汤可能通过调节肠道菌群的丰度和多样性影响肠道菌群,进而起到减轻肠道炎症和治疗溃疡性结肠炎的作用。

Effects of Shaoyao Decoction on Intestinal Flora of Ulcerative Colitis Rats Based on High-throughput Sequencing Technology

Objective To explore the effects of Shaoyao Decoction on the intestinal flora of rats with ulcerative colitis induced by TNBS;To discuss its mechanism of action in treating ulcerative colitis from the perspective of flora disorder.Methods Totally 50 SD rats were randomly divided into normal group,model group,antibiotic group,Shaoyao Decoction group,mesalazine group,with 10 rats in each group.A model of ulcerative colitis induced by 5%TNBS+50%ethanol.From the 8th day,the rats were treated with saline,antibiotics,Shaoyao Decoction,and mesalazine for 7 days.The histopathological changes of the colon were observed.ELISA was used to detect the content of inflammatory factors in the serum,and high-throughput sequencing technology was used to detect the changes of intestinal flora in rat feces.Results Compared with the normal group,the model group had severe colon damage,and the serum IL-6,IL-1β,and TNF-αlevels in rats significantly increased(P<0.01).Compared with the model group,the colon injury was alleviated in Shaoyao Decoction group,mesalazine group and antibiotic group,and the serum levels of IL-6,IL-1β,and TNF-αdecreased(P<0.01,P<0.05).In the model group,the abundance and diversity of the flora of rats decreased,and the abundance and diversity of intestinal flora increased after the treatment of Shaoyao Decoction and mesalazine.Compared with the model group,Phylum Firmicutes,Proteobacteria and the genus abundance of Clostridium,Oscillospira,Lactobacilli,Prevotella,Rumenococcus,Paraprevotella,and Roseburia increased in Shaoyao Decoction group and mesalazine group,while the abundance of Bacteroides phylum and Bacteroides genus decreased.Conclusion Intestinal flora is closely related to the secretion and expression of cytokines in the body.Shaoyao Decoction may affect the interacting flora by regulating the abundance and changes of the internal flora,and then play a role in reducing intestinal inflammation and treating ulcerative colitis.