昼夜节律紊乱对肠道微生态和粪便代谢组学影响的实验研究

Experimental study on the influence of circadian rhythm disturbance on intestinal microecology and fecal metabolites

目的探究昼夜节律紊乱对肠道微生态和粪便代谢物特征的影响。方法选取12只健康C57BL/6J小鼠(雄性,6~8周龄,18~25 g)作为研究对象,随机分为2组,每组6只。采用转换饲养环境光照节律的方法模拟昼夜节律变化,建立昼夜节律紊乱小鼠模型。4个月后,收集小鼠粪便和空肠内容物,采用16S rRNA测序法分析小鼠空肠和结肠内菌群的变化,采用靶向代谢组学分析粪便样品中代谢组学的变化。结果与昼夜节律正常组(Con组,n=6)比较,昼夜节律紊乱组(JL组,n=5)小鼠结肠菌群的丰度和多样性均降低,其中Bacteroidota门细菌丰度显著降低,Firmicutes门和Actinobacteria门细菌丰度均增高。粪便代谢组学结果显示,与Con组比较,JL组小鼠粪便中氨基酸水平显著降低(14.11%比30.04%,P<0.01);两组小鼠粪便中碳水化合物和短链脂肪酸水平的差异均无统计学意义(P均>0.05)。结论昼夜节律改变能够引起小鼠肠道微生态及粪便代谢物的变化。

Objective This study aims to explore the influence of circadian rhythm disorder on intestinal microecology changes and fecal metabolites characteristics.Methods Twelve healthy C57BL/6J mice(male,6 to 8 weeks old,18 to 25 g)were selected and randomly assigned to two groups with 6 mice in each group.The change of circadian rhythm was simulated by changing the light rhythm of the rearing environment,and the mouse model of circadian rhythm disorder was established.After 4 months,the stool samples and jejunal contents were collected.The jejunal contents and microbiota of fecal samples were analyzed by 16S ribosomal RNA sequencing.Targeted metabolomics was used to explore the differences of fecal metabolites between the two groups.Results Compared with the normal circadian rhythm group(Con group,n=6),the abundance and diversity of colonic flora of the mice in the circadian rhythm disorder group(JL group,n=5)are decreased,and the abundance of Bacteroidota is significantly decreased.The bacterial abundance of Firmicutes and Actinobacteria increased.Fecal metabolomics results show that compared with the Con group,the levels of amino acids in the feces of the mice in the JL group are significantly lower(14.11%versus 30.04%,P<0.01).There is no difference in the levels of carbohydrates and short-chain fatty acids in the feces of the two groups of mice.Conclusion Disturbances in the circadian rhythm can indeed trigger changes in the intestinal microecology and fecal metabolites in mice.