目的 基于网络药理学数据挖掘和实验验证探讨贯叶金丝桃（Hypericum perforatum L.）主要活性成分抗抑郁作用机制。 方法 通过文献检索和中药系统药理学数据库与分析平台等方法获取贯叶金丝桃主要活性成分，利用GeneCards数据库、STRING数据库、DAVID数据库进行网络药理学分析及GO富集分析和KEGG富集通路分析。利用Cytoscape 3.8.2软件构建贯叶金丝桃活性成分－靶点网络，筛选度值≥ 10、介数中心性≥ 0.01的节点进行拓扑分析。同时进行细胞实验和动物实验验证，将小胶质细胞（BV-2）分为正常组（Control）组、脂多糖（LPS）组、金丝桃素（HY）组以及钙调磷酸酶抑制剂（CNIS）组，利用免疫荧光法检测离子钙结合接头分子1（IBA-1）阳性表达和Western blot测定相关蛋白表达量。同时，将C57BL/6J小鼠随机分为Control组、抑郁（DP）组和HY组各15只，利用行为学检测黑白场实验（LDB）、旷场实验（OFT）、悬尾实验（TST）评估金丝桃素对慢性不可预见性应激（CUMS）诱导的DP样行为的改善作用，利用Western blot检测炎症因子肿瘤坏死因子α（TNF-α）和钙离子通路相关蛋白表达水平。 结果 筛选得到贯叶金丝桃中8个活性成分，预测到80个潜在的与抑郁相关的靶点；通过构建有效成分与交集基因可视化分析图发现HY在抗抑郁中发挥较为主要作用，再根据“成分-通路-靶点网络图”网络图发现在抗抑郁中关键的信号通路为钙离子信号通路。细胞实验结果表示，与Control组相比，LPS组IBA-1表达明显上升（P < 0.05）和促炎细胞因子及钙调蛋白（CaM）－钙调磷酸酶蛋白（CN）-活化T细胞因子（NFAT）钙信号通路的蛋白表达量显著升高（P < 0.05）；而与LPS组相比，HY组的IBA-1表达明显降低（P < 0.05）和TNF-α及CaM-CN-NFAT钙信号通路的蛋白表达显著降低（P < 0.05）。动物实验结果表示，与Control组相比，DP组小鼠OFT中央区域停留时间显著缩短（P < 0.05），LDB明暗箱穿梭次数显著减少（P < 0.05），TST不动时间显著延长（P < 0.05），促炎细胞因子（TNF-α）及钙信号通路的蛋白表达水平显著升高（P < 0.05），HY组逆转了上述结果。 结论 通过网络药理分析初步揭示了贯叶金丝桃抗抑郁活性成分作用机制并进行了体内外验证，为进一步的药理机制研究提供了理论依据。

Objective To explore the antidepressant mechanism of the main active ingredients of Hypericum perforatum L. through network pharmacology data mining and experimental validation. Methods The main active ingredients of Hypericum perforatum L. were retrieved from the literature and the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform. Network pharmacology analysis， GO enrichment analysis， and KEGG pathway enrichment analysis were carried out using the GeneCards， STRING， and DAVID databases. The active ingredient–target network of Hypericum perforatum was constructed with Cytoscape 3.8.2 software. Nodes with a degree value of at least 10 and betweenness centrality of at least 0.01 were selected for topological analysis. Meanwhile， cellular and animal experiments were conducted to validate the results. Microglial cells （BV-2） were divided into four groups： Control， lipopolysaccharide （LPS）， hypericin （HY）， and calcineurin inhibitor （CNIS）. Immunofluorescence was employed to detect the positive expression of ionized calcium-binding adapter molecule 1 （IBA-1）， and Western blot was utilized to determine the expression levels of related proteins. Additionally， C57BL/6J mice were randomly assigned to three groups： Control， depression （DP）， and HY groups （n = 15 per group）. Behavioral tests， such as the light-dark box （LDB）， open field test （OFT）， and tail suspension test （TST）， were used to assess the ameliorative effect of hypericin on chronic unpredictable mild stress （CUMS）-induced depression-like behaviors. Western blot was also used to measure the expression levels of the inflammatory factor tumor necrosis factor-α （TNF-α） and proteins related to the calcium signaling pathway. Results Eight active ingredients were screened from Hypericum perforatum， and 80 potential depression-related targets were predicted. Through visualization analysis of the active ingredients and intersecting genes， it was found that HY plays a major role in the antidepressant effect. The “ingredient?pathway?target” network analysis indicated that the calcium signaling pathway was a key pathway. Cell experiments demonstrated that， in comparison with the Control group， the LPS group showed a significant increase in IBA-1 expression （P < 0.05） and a significant increase in the protein expression of pro-inflammatory cytokines and the CaM-CN-NFAT calcium signaling pathway （P < 0.05）. In contrast， compared with the LPS group， the HY group presented a significant decrease in IBA-1 expression （P < 0.05） and a significant decrease in the protein expression of TNF-α and the CaM-CN-NFAT calcium signaling pathway （P < 0.05）. Animal experiments revealed that， when compared with the Control group， the DP group had a significant shortening of the central zone time in the OFT （P < 0.05）， a significant reduction in the number of transitions in the LDB （P < 0.05）， a significant prolongation of immobility time in the TST （P < 0.05）， and a significant increase in the protein expression of TNF-α and calcium signaling pathway proteins （P < 0.05）. All these changes were reversed in the HY group. Conclusion This study has preliminarily revealed the mechanism of antidepressant active ingredients from Hypericum perforatum L. via network pharmacology analysis combined with in vivo and in vitro validation， thus providing a theoretical basis for further pharmacological research.