目的 基于网络药理学及动物实验，探讨双夏汤对于慢性不可预知温和应激刺激（CUMS）抑郁样大鼠行为的改善作用及分子机制。方法 采用网络药理学方法收集双夏汤的活性成分和治疗抑郁症的潜在靶点，筛选核心网络靶点。通过动物实验对网络药理学结果进行实验验证，具体方法为连续3周通过CUMS建立大鼠抑郁样模型，给予双夏汤干预2周，给药结束后进行行为学测试，通过旷场实验、强迫游泳实验评估抑郁行为，ELISA法检测大鼠血清白细胞介素（IL）-6水平，蛋白免疫印迹法检测各组大鼠海马组织中低氧诱导因子-1α（HIF-1α）、cAMP反应元件结合蛋白（CREB）表达水平，及其p-蛋白激酶B1（Akt）、p-糖原合成酶激酶3β（GSK3B）蛋白表达水平。结果 网络药理学分析获得245个双夏汤抗抑郁症的潜在靶点，蛋白相互作用（PPI）网络分析发现双夏汤可能通过调控GSK3B、Akt1、磷脂酰肌醇-4,5-二磷酸肌醇-3-激酶（PIK3CA）等靶蛋白发挥抗抑郁作用，涉及神经活性配体与受体的相互作用、HIF-1等信号通路。与模型组比较，双夏汤组大鼠体质量明显上升，强迫游泳测试及旷场测试中大鼠不动时间明显缩短（P＜0.05、0.001），血清IL-6水平明显下降（P＜0.01），HIF-1α、CREB、p-Akt、p-GSK3B蛋白表达明显降低，GSK3B蛋白表达明显升高。结论 双夏汤对于CUMS大鼠的抑郁样行为具有改善作用，其机制可能是通过Akt/GSK3B和HIF-1α/CREB信号通路调节机体昼夜节律及海马组织中葡萄糖稳态而改善海马神经元可塑性。

Objective To investigate the ameliorative effect and molecular mechanism of Shuangxia Decoction on the behaviour of chronic unpredictable mild stress stimulation (CUMS)-like depressed rats based on network pharmacology and animal experiments. Methods A network pharmacology approach was used to collect the active ingredients and potential targets of Shuangxia Decoction for the treatment of depression and to screen the core network targets. The network pharmacology results were experimentally verified through animal experiments, as follows: depression-like modeling in rats by CUMS for 3 consecutive weeks, Shuangxia Decoction and fluoxetine were given to intervene for 2 weeks, and behavioral tests were conducted at the end of drug administration to assess the depressive behaviors through the absent-field experiment and forced-swimming experiment, the serum IL-6 level of the rats was detected by the ELISA method, and the Western blotting was used to detect the protein expression levels of HIF-1α and CREB, as well as the expression levels of p-Akt and p-GSK3B proteins in the hippocampal tissues of rats in each group. Results Network pharmacological analysis obtained 245 potential targets of Shuangxia Decoction for antidepression. PPI network analysis revealed that Shuangxia Decoction might exert antidepressant effects by regulating target proteins such as GSK3B, Akt1 and PIK3CA. It involves neuroactive ligand- receptor interaction, HIF-1 and other signaling pathways. The experimental results showed that compared with the model group, the body mass of rats in the Shuangxia Decoction group were increased significantly, and immobility time of rats in the forced swimming test and open field test was shortened significantly (P < 0.05, 0.001), serum IL-6 was significantly were decreased (P< 0.01). And the protein expression of HIF-1α, CREB, p-Akt, and p-GSK3B was significantly reduced, and that of GSK3B was significantly elevated. Conclusion Shuangxia Decoction can improve the depression-like behavior of CUMS rats, and its mechanism may be to improve the plasticity of hippocampal neurons through the regulation of circadian rhythm and glucose homeostasis in hippocampal tissues through AKT/GSK3B and HIF-1α/CREB signaling pathways.

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国家重点研发计划项目（2017YFC1702700）;山东省中医药科技项目（2021Q077）;山东省高等学校青创科技支持计划（2023RW093）