溃疡性结肠炎诊治潜在靶点探讨——基于肠道菌群与代谢组学联动分析  

Identification of potential targets for diagnosis and treatment of ulcerative colitis based on linkage analysis of intestinal flora and metabolomics

作　　者：于官正 李鸿 涂星 马焌茹 王楚秀 聂娟 YU Guanzheng;LI Hong;TU Xing;MA Junru;WANG Chuxiu;NIE Juan(Chinese Medicinal Materials Products Quality Supervision and Inspection Center in Wuling Mountainous Area,Enshi,Hubei 445000,China;不详)

机构地区：[1]湖北民族大学武陵山中药材检验检测中心,湖北恩施445000 [2]湖北民族大学医学部 [3]云南中医药大学第一临床医学院

出　　处：《中国微生态学杂志》2024年第12期1406-1416,共11页Chinese Journal of Microecology

基　　金：国家自然科学基金青年科学基金(82304829);恩施州科技项目技术支撑类重点项目(D20220065)。

摘　　要：目的基于UPLC-QE-MS和16S rRNA基因测序技术分析溃疡性结肠炎(UC)小鼠肠道菌群及内源性代谢物之间的联动系统,挖掘关键生物标志物,探讨UC治疗的潜在靶点。方法将雄性C57小鼠(体重18~22 g,8周龄)随机分为正常组与模型组,每组10只,模型组小鼠自由饮用2.5%葡聚糖硫酸钠诱导UC模型。记录小鼠体重变化率和DAI评分;测量结肠长度;HE染色和AB-PAS染色观察肠组织病理变化;ELISA法检测血清IL-1β与IL-10水平;非靶向代谢组学技术和16S rRNA基因测序技术联合分析小鼠内源性代谢物和肠道菌群的变化,挖掘潜在生物标志物。结果与正常组比较,模型组小鼠体重变化率显著下降(t=21.727)、DAI评分显著上升(t=−17.376)、结肠长度显著缩短(t=8.514),均P<0.05;肠组织明显形成溃疡,伴随隐窝与绒毛消融、炎性细胞浸润、黏液层缺失;血清IL-1β水平显著升高(t=−10.744)、IL-10水平显著降低(t=10.191),均P<0.05。肠道菌群分析结果表明,模型组小鼠肠道菌群多样性下降,显著差异菌群有Lachnospiraceae_NK4A136_group、Bacteroides、Muribaculaceae等。代谢组学挖掘出83个UC相关潜在生物标志物,以甘油磷脂、氨基酸、类花生酸成分为主,涉及精氨酸的生物合成等23条代谢通路。联合分析结果表明差异代谢物与差异菌属存在显著关联性。结论UC的发生发展伴随肠道菌群多样性下降与代谢紊乱。调节肠道菌群多样性、平衡氨基酸合成、甘油磷脂代谢和类花生酸代谢可能是诊疗UC的潜在靶点。Objective To explore the interrelationship between intestinal flora and endogenous metabolites in mice with ulcerative colitis(UC),identify key biomarkers and explore potential therapeutic targets for UC using UPLC-QE-MS and 16S rRNA technology.Methods Male C57 mice(Weight 18 to 22 g,8 weeks old)were randomly allocated into the normal group(n=10)and the model group(n=10).The model group freely consumed 2.5%dextran sulfate(DSS)to induce the UC model.The change in body weight and the DAI score of mice were recorded.Colon length was measured.The pathological alterations of intestinal tissue were observed through HE and AB-PAS staining.Serum levels of IL-1βand IL-10 were detected with ELISA.Non-targeted metabolomics technology and 16S rRNA gene sequencing technology were combined to analyze the alterations of endogenous metabolites and intestinal microbiota in mice to explore potential biomarkers.Results In comparison with the normal group,the weight change in the model group significantly decreased(t=21.727,P<0.05), the DAI score significantly elevated (t=−17.376, P<0.05), and the colon length was significantly shortened(t=8.514, P<0.05). The intestinal tissue conspicuously developed ulcers, accompanied by the disintegration of crypts andvilli, infiltration of inflammatory cells, and the absence of mucus layer. The level of IL-1β in the serum significantly increased(t=−10.744, P<0.05), while that of IL-10 significantly reduced (t=10.191, P<0.05). The findings of the intestinalmicrobiome demonstrated that the diversity of the microbiome in the model group was diminished, and there were significantdifferences in the bacterial populations such as Lachnospiraceae_NK4A136_group, Bacteroides, and Muribaculaceae,etc. Metabolomics identified 83 potential biomarkers related to UC, with glycerophospholipids, arachidonic acid, andamino acid components as the main constituents, involving Arginine biosynthesis and other 23 metabolic pathways. Thecombined analysis results indicated that there was a significant correlation b

关 键 词：溃疡性结肠炎 肠道菌群 代谢组学 16S rRNA基因测序 UPLC-QE-MS 

分 类 号：R363.2[医药卫生—病理学]