基于网络药理学探究苓桂术甘汤治疗抑郁症的作用机制及实验验证

Mechanism of Linggui Zhugan Decoction on Depression based on Network Pharmacology and Experimental Validation

目的基于网络药理学和实验验证探究复方苓桂术甘汤治疗抑郁症的作用机制。方法运用网络药理学技术筛选苓桂术甘汤中与抑郁症相关的靶点,获取潜在作用靶点后构建PPI网络,利用Metascape平台对潜在作用靶点进行GO和KEGG通路富集分析,采用Cytoscape3.7.1软件构建苓桂术甘汤成分-抑郁症靶点-主要通路网络,并利用分子对接技术验证上述结果。采用皮下注射皮质酮的方法制备抑郁小鼠模型,给予丙咪嗪与苓桂术甘汤干预,通过糖水偏好实验、悬尾实验、强迫游泳实验、开野实验观察各组小鼠抑郁样行为,尼氏染色法检测小鼠海马尼氏小体数量变化。结果网络药理学分析表明,苓桂术甘汤治疗抑郁症的核心成分为槲皮素、美迪紫檀素、β-谷甾醇、山柰酚、刺芒柄花素等,核心靶点有ALB、IL6、AKT1、MAPK3、FOS、VEGFA、TNF、CASP3、NOS3、PTGS2等。苓桂术甘汤发挥抗抑郁作用的主要通路有神经活性配体-受体相互作用信号通路、钙信号通路、癌症通路、糖尿病并发症中的AGE-RAGE信号通路、cAMP信号通路等。苓桂术甘汤发挥抗抑郁作用的主要通路有神经活性配体-受体相互作用信号通路、钙信号通路、癌症通路、糖尿病并发症中的AGE-RAGE信号通路、cAMP信号通路等。体内实验结果表明与皮质酮模型小鼠相比,给予苓桂术甘汤治疗的小鼠糖水偏好率明显增加(P<0.01),强迫游泳及悬尾不动时间均显著减少(P<0.01),开野实验中移行总距离和中央移行距离显著增加(P<0.05,P<0.01),小鼠海马神经元数量增加(P<0.01)。结论苓桂术甘汤能够通过"多成分、多靶点、多通路"治疗抑郁症,改善抑郁小鼠海马内神经元损伤,缓解抑郁样行为。

OBJECTIVE To explore the mechanism of Linggui Zhugan decoction(LGZGD)on depression through network pharmacology and experimental verification.METHODS The targets related to depression in Linggui Zhugan decoction were screened by network pharmacology technology,and the PPI network was constructed after the potential action targets were obtained.The Metascape platform was used to conduct enrichment analysis of the GO and KEGG pathways of the potential action targets.Cytoscape 3.7.1 software was used to construct the Linggui Zhugan decoction component-depression target-main pathway network.The above results were verified by molecular docking technique.The model mice were subcutaneously injected with corticosterone,Imipramine and Linggui Zhugan decoction were administered to intervene.The depression-like behavior of the mice in each group was observed by sucrose preference test(SPT),tail suspension test(TST),forced swimming test(FST)and open field test(OFT),and the morphology and number of Nissl was observed by Nissl staining.RESULTS Network pharmacological analysis showed that the core components of Linggui Zhugan decoction in the treatment of depression were quercetin,medicarpin,β-sitosterol,kaempferol,formononetin,and the core targets were ALB,IL6,AKT1,MAPK3,FOS,VEGFA,TNF,CASP3,NOS3,PTGS2,etc.The main pathways in which Linggui Zhugan decoction played an antidepressant role include neuroactive ligand-receptor interaction,calcium signaling pathway,pathways in cancer,AGE-RAGE signaling pathway in diabetic complications and cAMP signaling pathway.In vivo experiment results showed that compared with the corticosterone-induced mice,the sucrose preference rate of the mice treated with Linggui Zhugan decoction was significantly increased(P<0.01),the forced swimming and tail suspension immobility time were significantly reduced(P<0.01),the total migration distance and central migration distance were significantly increased in the open field test(P<0.05 or P<0.01),and the number of hippocampal neurons was increased(P<0.01).CONCLUSION Linggui Zhugan decoction can treat depression through"multi-component,multi-target and multi-pathway",improve neuron damage in hippocampus of depressed mice,and alleviate depressive-like behavior.