基于神经内分泌激素和代谢组学关联分析的复方柴归方抗抑郁作用机制研究

Anti-depressive mechanism of Fufang Chaigui prescription based on neuroendocrine hormone and metabolomic correlation analysis

为了阐明复方柴归方的抗抑郁效果及作用机制,并考察其对神经内分泌功能的影响,将SD大鼠进行为期28 d的慢性温和不可预知应激(chronic unpredictable mild stress,CUMS)造模,造模同时给予复方柴归方干预,实验期间观察大鼠行为学变化。用酶联免疫吸附法(ELISA)测定大鼠血清中与下丘脑-垂体-肾上腺皮质轴(HPAA)、下丘脑-垂体-甲状腺轴(HPTA)以及下丘脑-垂体-性腺轴(HPGA)相关的代表性激素浓度。并用1H-NMR代谢组学技术分析大鼠血清内源性代谢产物的变化规律,寻找出潜在的生物标志物。结果显示复方柴归方能明显改善CUMS模型大鼠的行为活动,回调CUMS引起血清中的促肾上腺皮质激素(ACTH)、皮质酮(CORT)、三碘甲状腺原氨酸(T3)浓度升高,促甲状腺激素(TSH)和睾酮(TESTO)浓度的降低,并且调节造模引起的内源性代谢产物乳酸、α-葡萄糖、胆碱、N-乙酰糖蛋白、氧化三甲胺、亮氨酸含量变化,使其趋近于正常对照组。关联分析发现HPTA与糖代谢、氨基酸代谢、胆碱代谢均有一定联系,HPAA与糖代谢和氨基酸代谢相关,而HPGA与糖代谢有关。表明复方柴归方抗抑郁效果明显,其作用机制可能涉及神经内分泌功能的调节以及糖代谢、氨基酸代谢、胆碱代谢等途径的调控。

To elucidate the anti-depressive effect of Fufang Chaigui prescription and its mechanism and investigate its effect on neuroendocrine hormone,rats were included into a chronic unpredictable mild stress（ CUMS） model for 28 d,and drugs were administered at the same time. During the period,rats＇ behaviors were observed and the blood was collected by using ELISA to determine representative hormone concentrations of HPAA,HPTA and HPGA. The changes in endogenous metabolites were analyzed by using1 H NMR metabolomics to seek the potential biomarkers. Results showed Fufang Chaigui prescription could improve the behaviors of CUMS rats obviously,increase contents of ACTH,CORT,T3 and decrease contents of TSH and TESTO and regulate the levels of lactate,α-glucose,choline,N-acetylglycoprotein,trimethylamine oxide and leucine to get closer to the contents of control group. The results of correlation analysis indicated that HPTA was associated with glycometabolism,amino acid metabolism and choline metabolism.And HPAA was related to glycometabolism and amino acid metabolism. However,HPGA was only correlated with glycometabolism. In conclusion,Fufang Chaigui prescription could show an obvious anti-depressive effect and its underlying mechanism might involve regulations of neuroendocrine function and pathways of glycometabolism,amino acid metabolism and choline metabolism.