肠易激综合征小鼠粪便代谢组学及炒白术干预作用研究

Fecal Metabolic Profiling in a Murine Model of Irritable Bowel Syndrome and the Intervention Role of Prepared Atractylodes Macrocephala

目的通过检测肠易激综合征(IBS)小鼠粪便代谢物产物,以期反映肠道菌群代谢变化,并以中药炒白术作为干预药物,观察药物对肠道菌群代谢的影响。方法 18只雌性C57BL/6小鼠随机分为正常对照组、模型组、白术组,每组6只。除正常对照组外,其余小鼠采用避水应激法(WAS)建立IBS小鼠模型。造模10天后,白术组给予炒白术煎剂灌胃,对照组及模型组给予等体积生理盐水灌胃,持续7天。观察各组小鼠给药后腹肌收缩反射(AWR)评分及粪便性状变化。收集并计数实验开始第0、5、10及17天的小鼠粪便颗粒数。运用气相色谱-质谱(GC-MS)检测以及多变量统计学数据分析,比较正常对照组和模型组,白术组和模型组的粪便代谢产物。结果正常对照组和模型组存在粪便代谢产物差异,共鉴定出14种差异代谢物。苯丙氨酸代谢是最相关代谢通路。白术组较模型组小鼠粪便成形,颗粒数减少(P<0.05),AWR评分降低(P<0.05,P<0.01)。白术组和模型组共鉴定出16种差异代谢物。苯丙氨酸代谢、烟酸和烟酰胺代谢及泛酸和辅酶A生物合成是最相关代谢通路。结论 IBS小鼠存在粪便代谢产物及代谢通路改变,提示肠道菌群代谢紊乱。中药炒白术可改善IBS小鼠内脏敏感性及粪便性状,可能与影响肠道菌群代谢相关。

Objective To observe the metabolic changes of gut microbiota by detecting fecal metabolites of mice with irritable bowel syndrome(IBS), and to observe the effects of prepared Atractylodes Macrocephala(AM), as an interventive drug, on intestinal flora metabolism. Methods Eighteen female C57BL/6 mice were randomly divided into the normal control group, the model group, and the AM group, 6 female in each group. IBS mice model was established using water avoidance stress(WAS) in mice except the normal control group. After 10 days modeling, AM decoction was administered to rats in the AM group by gastrogavage, and equal volume of normal saline was administered to rats in the control group and the model group by gastrogavage. All medication lasted for 7 successive days. Changes of abdominal withdrawal reflex(AWR) and fecal traits were observed after drug administration. The numbers of fecal particles were collected counted on day 0, 5,10, and 17, respectively. Gas chromatography-mass spectrometry(GC-MS) and multivariate statistical data analysis were used to compare the fecal samples between the normal control group and the model group, and between the AM group and the model group. Results There were differences in fecal metabolites between the normal control group and the model group. A total of 14 differential metabolites were identified from these two groups. And phenylalanine metabolism was the most significantly associated metabolic pathway. The fecal traits and defecation were improved(P<0.05), and the visceral hypersensitivity of AWR score decreased(P<0.05, P<0.01) more obviously in the AM group than in the model group. A total of 16 differential metabolites were identified from the AM group and the model group. Phenylalanine, nicotinic acid, and nicotinamide, pantothenate and CoA biosynthesis were the most significantly associated metabolic pathways. Conclusions Changes in metabolites and metabolic pathways existed in feces of IBS mice, indicating abnormal metabolism of intestinal flora. Prepared AM alleviated visceral sensitivity and feces traits, which might be associated with influencing the metabolism of intestinal flora.