目的 基于網(wǎng)絡(luò)藥理學(xué)及動(dòng)物實(shí)驗(yàn)，探討雙夏湯對于慢性不可預(yù)知溫和應(yīng)激刺激（CUMS）抑郁樣大鼠行為的改善作用及分子機(jī)制。方法 采用網(wǎng)絡(luò)藥理學(xué)方法收集雙夏湯的活性成分和治療抑郁癥的潛在靶點(diǎn)，篩選核心網(wǎng)絡(luò)靶點(diǎn)。通過動(dòng)物實(shí)驗(yàn)對網(wǎng)絡(luò)藥理學(xué)結(jié)果進(jìn)行實(shí)驗(yàn)驗(yàn)證，具體方法為連續(xù)3周通過CUMS建立大鼠抑郁樣模型，給予雙夏湯干預(yù)2周，給藥結(jié)束后進(jìn)行行為學(xué)測試，通過曠場實(shí)驗(yàn)、強(qiáng)迫游泳實(shí)驗(yàn)評(píng)估抑郁行為，ELISA法檢測大鼠血清白細(xì)胞介素（IL）-6水平，蛋白免疫印跡法檢測各組大鼠海馬組織中低氧誘導(dǎo)因子-1α（HIF-1α）、cAMP反應(yīng)元件結(jié)合蛋白（CREB）表達(dá)水平，及其p-蛋白激酶B1（Akt）、p-糖原合成酶激酶3β（GSK3B）蛋白表達(dá)水平。結(jié)果 網(wǎng)絡(luò)藥理學(xué)分析獲得245個(gè)雙夏湯抗抑郁癥的潛在靶點(diǎn)，蛋白相互作用（PPI）網(wǎng)絡(luò)分析發(fā)現(xiàn)雙夏湯可能通過調(diào)控GSK3B、Akt1、磷脂酰肌醇-4,5-二磷酸肌醇-3-激酶（PIK3CA）等靶蛋白發(fā)揮抗抑郁作用，涉及神經(jīng)活性配體與受體的相互作用、HIF-1等信號(hào)通路。與模型組比較，雙夏湯組大鼠體質(zhì)量明顯上升，強(qiáng)迫游泳測試及曠場測試中大鼠不動(dòng)時(shí)間明顯縮短（P＜0.05、0.001），血清IL-6水平明顯下降（P＜0.01），HIF-1α、CREB、p-Akt、p-GSK3B蛋白表達(dá)明顯降低，GSK3B蛋白表達(dá)明顯升高。結(jié)論 雙夏湯對于CUMS大鼠的抑郁樣行為具有改善作用，其機(jī)制可能是通過Akt/GSK3B和HIF-1α/CREB信號(hào)通路調(diào)節(jié)機(jī)體晝夜節(jié)律及海馬組織中葡萄糖穩(wěn)態(tài)而改善海馬神經(jīng)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.

R285

國家重點(diǎn)研發(fā)計(jì)劃項(xiàng)目（2017YFC1702700）;山東省中醫(yī)藥科技項(xiàng)目（2021Q077）;山東省高等學(xué)校青創(chuàng)科技支持計(jì)劃（2023RW093）