白术多糖对抑郁症小鼠动物模型的作用及机制研究

Effect and Mechanism of Atractylodes macrocephala polysaccharide on CUMS Mouse Models

目的整合代谢组学技术和肠道菌群分析,探讨白术多糖(Atractylodes macrocephala polysaccharide,AMP)改善慢性不可预知性刺激(chronic unpredictable mild stress,CUMS)抑郁小鼠模型的作用机制。方法建立CUMS抑郁小鼠模型,AMP低、中、高剂量治疗组给药剂量分别为0.0625、0.125、0.250 g·kg^(-1),氟西汀组为0.015 g·kg^(-1),对照组和模型组给予0.9%氯化钠溶液,所有组给药量均为5 mL·kg^(-1)·d^(-1)。酶联免疫法测定小鼠大脑中五羟色胺(5-hydroxytryptamine,5-HT)的含量,结合糖水偏好试验结果检验CUMS抑郁模型;采用16S rDNA扩增子测序技术对各组小鼠粪便中肠道菌群测序分析,液相色谱-质谱联用(LC-MS)技术对小鼠血清进行非靶向代谢组学测定。结果与对照组比较,CUMS模型小鼠糖水偏好率明显降低,小鼠大脑内的5-HT含量显著降低(P<0.01),提示CUMS抑郁模型建立成功。AMP治疗后,各组小鼠糖水偏好率和5-HT含量均增加(P<0.01);CUMS小鼠肠道菌群丰度具有回调效果;血清代谢物发生显著改变,58种代谢物显著回调;斯皮尔曼(Spearman)相关性分析显示,肠道菌群的改变与代谢物水平变化显著性相关。结论AMP通过调节肠道菌群稳态,上调厚壁菌门与拟杆菌门的比值(F-B),进而调控代谢途径,对CUMS抑郁模型小鼠发挥干预效用。

OBJECTIVE To explore the mechanism of Atractylodes macrocephala polysaccharide(AMP)in improving chronic unpredictable mild stress(CUMS)depression in mice by integrating metabolomics technology and gut microbiota analysis.METHODS CUMS depression mouse model was established.Low,medium and high dose AMP treatment were given at 0.0625,0.125,0.250 g·kg^(-1),respectively,fluoxetine was given at the dosage of 0.015 g·kg^(-1),control group and model group were given 0.9%saline solution,and all groups were given at 5 mL·kg^(-1)·d^(-1) of drug.The content of 5-hydroxytryptamine(5-HT)in the brain of mice was determined by enzyme-linked immunosorbent assay(ELISA),and the CUMS depression model was tested by combining sucrose preference test(SPT)results.The 16S rDNA amplicon sequencing technology was used to analyze the gut microbiota in the feces of mice in each group.LC-MS technology was used to perform non-targeted metabolomics determination of mouse serum.RESULTS Compared with the control group,the sucrose preference rate of CUMS model mice was significantly lower,and the content of 5-HT in the brain of mice was significantly reduced(P<0.01),indicating that the CUMS depression model was successfully established.After AMP treatment,the sucrose preference rate and 5-HT content of mice in each group increased(P<0.01),the gut microbiota of CUMS mice had a regulatory effect,serum metabolites were significantly changed,and 58 metabolites were significantly adjusted.Spearman correlation analysis showed that the changes in gut microbiota were significantly associated with the changes in metabolite levels.CONCLUSION AMP exerts intervention effects on CUMS depression model mice by regulating the stability of gut microbiota,upregulating the F-B ratio,and thereby regulating metabolic pathways.