【摘要】 目的：确定复方萱草提取物浓缩液（CHEC）中各草药成分的最佳配比，在斑马鱼和小鼠抑郁模型中评价其抗抑郁作用，并利用网络药理学和分子生物学方法，特别关注磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B(AKT)/脑源性神经营养因子（BDNF）信号通路，阐明其潜在机制。方法：使用利血平诱导的斑马鱼抑郁模型筛选最佳CHEC配方。为了评估神经炎症和单胺类神经递质水平，使用酶联免疫吸附测定试剂盒检测斑马鱼脑组织中白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α、多巴胺和5-羟色胺的水平。通过液相色谱-串联质谱法鉴定活性成分，并使用Swiss Target Prediction数据库预测潜在靶点。构建了中药-活性成分-靶点网络和蛋白质-蛋白质相互作用网络，随后进行基因本体论和京都基因与基因组百科全书富集分析以探索潜在机制。随后，通过慢性束缚应激联合腹腔注射脂多糖建立小鼠抑郁模型。通过行为学测试、单胺类神经递质和炎症细胞因子的定量检测、苏木精-伊红染色海马神经元组织病理学检查以及PI3K/AKT/BDNF通路相关蛋白的蛋白质印迹分析，评估CHEC的抗抑郁效果。结果：行为学评估表明，CHEC能显著减轻慢性束缚应激小鼠的抑郁样行为。CHEC治疗降低了促炎细胞因子水平，恢复了单胺类神经递质浓度。组织学分析显示，海马神经元损伤得到减轻。此外，CHEC显著调节了抑郁样小鼠海马中的PI3K/AKT/BDNF信号通路。结论：CHEC通过抑制炎症细胞因子（包括肿瘤坏死因子-α、IL-6和IL-1β）、提高多巴胺和BDNF水平以及调节PI3K/AKT/BDNF信号通路，发挥抗抑郁样作用。这些发现为CHEC治疗抑郁症的潜力提供了机制性证据。更多还原

【Abstract】 Objective: To determine the optimal dosing ratio of the herbal components within compound Hemerocallis citrina Baroni(H. citrina) extract concentrate(CHEC), evaluate its antidepressant effects in zebrafish and mouse models of depression, and elucidate the underlying mechanisms with a particular focus on the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/brain-derived neurotrophic factor(BDNF) signaling pathway using network pharmacology and molecular biology approaches.Methods: The optimal CHEC formulation was screened using a zebrafish model of reserpine-induced depression. To assess neuroinflammation and monoamine neurotransmitter levels, the levels of interleukin(IL)-1β, tumor necrosis factor(TNF)-α, dopamine, and 5-hydroxytryptamine in zebrafish brain tissues were measured using enzyme-linked immunosorbent assay kits. Active constituents were identified by liquid chromatography-tandem mass spectrometry and potential targets were predicted using the Swiss Target Prediction database. A traditional Chinese medicine–active ingredient–target network and a protein–protein interaction network were constructed, followed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses to explore potential mechanisms. A mouse model of depression was subsequently established using chronic restraint stress(CRS) combined with intraperitoneal lipopolysaccharide injection. The antidepressant effects of CHEC were assessed through behavioral tests, quantification of monoamine neurotransmitters and inflammatory cytokines,histopathological examination of hippocampal neurons using hematoxylin and eosin staining, and Western blot analysis of PI3K/AKT/BDNF pathway-related proteins.Results: CHEC significantly alleviated depressive-like behaviors in CRS mice, as demonstrated by behavioral assessments. Treatment reduced levels of pro-inflammatory cytokines and restored monoamine neurotransmitter concentrations. Histological analysis revealed attenuation of hippocampal neuronal damage. Furthermore, CHEC significantly modulated the PI3K/AKT/BDNF signaling pathway in the hippocampus of depressive-like mice.Conclusion: CHEC exerts antidepressant-like effects by suppressing inflammatory cytokines, including tumor necrosis factor-alpha, IL-6, and IL-1β; enhancing dopamine and BDNF levels; and regulating the PI3K/AKT/BDNF signaling pathway. These findings provide mechanistic evidence supporting the therapeutic potential of CHEC for depression.更多还原