目的 运用血清代谢组学与网络药理学技术挖掘疏肝和胃汤(Shuganheweitang，SGHWT)治疗抑郁症的相关代谢通路和作用靶点，探究疏肝和胃汤抗抑郁的作用机制。方法 将40只SD雄性大鼠随机选择10只为正常对照组，剩余30只大鼠利用慢性不可预知温和刺激(Chronic unpredictable mild stress，CUMS)复刻抑郁大鼠模型，分为模型组、疏肝和胃汤组(7。34 g·kg-1·d-1)和氟西汀组(1。58 mg·kg-1·d-1)，每组10只，连续造模和灌胃共6周。采用超高效液相色谱-四级杆-飞行时间质谱法(Ultra-performance liquid chromatography-quantitative time-of-flight tandem mass spectrometry，UPLC-Q-TOF-MS/MS)对大鼠血清中内源性差异代谢物进行检测。利用网络药理学方法筛选疏肝和胃汤中的核心成分和关键靶点信息。最后借助Cytoscape软件对差异代谢物及蛋白靶点进行联合分析，运用ELISA实验检测各组大鼠血清中花生四烯酸(Arachidonic acid，AA)、肿瘤坏死因子-α(Tumor necrosis factor-α，TNF-α)、白细胞介素-6(Interleukin-6，IL-6)和环氧合酶(Cyclooxygenase，COX-2)的含量。结果 从血清代谢组中共鉴定得到47个差异代谢物，疏肝和胃汤可回调其中18个差异代谢物，通路富集分析发现主要涉及初级胆汁酸的生物合成、花生四烯酸代谢(impact>0。1)。利用网络药理学手段筛选出疏肝和胃汤71种活性成分和47个关键靶点蛋白，联合分析中花生四烯酸及相关靶点所占比重最大。经过验证发现，与对照组相比，模型组血清中AA(P<0。05)、IL-6(P<0。01)和TNF-α(P<0。01)的含量均显著升高。与模型组相比，疏肝和胃汤组血清中IL-6和TNF-α的含量均显著降低(P<0。01)。结论 疏肝和胃汤有效组分治疗抑郁症具有多靶点和多通路的特性，其作用机制可能与疏肝和胃汤参与调控花生四烯酸代谢途径，抑制炎症因子水平，进而改善慢性应激诱导的抑郁样症状相关。
Exploring the Mechanism of Shuganheweitang in Antidepressant Action based on the Integrated Strategy of Serum Metabolomics and Network Pharmacology
Objective Serum metabolomics combined with network pharmacology were used to explore the related metabolic pathways and action targets of Shuganheweitang(SGHWT)in the treatment of depression,and to explore the anti-depression mechanism of SGHWT.Methods Ten of the 40 SD male rats were randomly selected as normal control group,and the remaining 30 rats were reproduced as depressed rat model by chronic unpredictable mild stress(CUMS).They were divided into model group,SGHWT group(7.34 g·kg-1·d-1)and fluoxetine group(1.58 mg·kg-1·d-1),with 10 animals in each group and ontinuous modeling and gavage lasted for 6 weeks.The endogenous differential metabolites in rat serum were detected by UPLC-Q-TOF-MS/MS.The core components and key target information of SGHWT were selected by network pharmacology.Finally,combined analysis of differential metabolites and protein targets was carried out by Cytoscape software.Serum contents of arachidonic acid(AA),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and cyclooxygenase(COX-2)in each group were detected according to the analysis results.Results A total of 47 different metabolites were identified from the serum metabolome,18 of which could be recovered by SGHWT.Through pathway enrichment analysis,it was found that it was mainly involved in arachidonic acid metabolism and primary bile acid biosynthesis.71 active ingredients and 47 key target proteins of SGHWT were screened by network pharmacological means,and the proportion of arachidonic acid and related targets was the largest in the combined analysis.The ELISA results showed that compared with the control group,the serum contents of AA(P<0.05),IL-6(P<0.01)and TNF-α(P<0.01)in model group were significantly increased.Compared with model group,the contents of IL-6 and TNF-α in SGHWT group were significantly decreased(P<0.01).Conclusion The effective components of SGHWT have the characteristics of multi-target and multi-pathway in the treatment of depression.Further,the mechanism of action may be related to the fact that SGHWT is involved in regulating the metabolic pathway of arachidonic acid,inhibiting the level of inflammatory factors,and then improving the depression-like symptoms induced by chronic stress.
万方数据
