目的 探究柴胡疏肝散治療抑郁癥的藥效物質及作用機制并進行實驗驗證。方法 應用超高效液相色譜-四極桿飛行時間質譜聯(lián)用（UPLC-Q-TOF-MS/MS）技術對柴胡疏肝散化學成分進行分析；采用中藥系統(tǒng)藥理學數(shù)據(jù)庫及分析平臺（TCMSP）和SwisTargetPrediction數(shù)據(jù)庫獲取并篩選成分靶點；運用Limma包和hclust函數(shù)對GSE19738基因芯片數(shù)據(jù)進行差異基因分析和加權基因共表達網(wǎng)絡（WGCNA）分析，獲取抑郁癥相關靶點，并與成分靶點取交集。繪制成分-靶點網(wǎng)絡圖，計算度值，篩選藥效物質。交集基因通過基因本體（GO）功能富集分析和京都基因與基因組百科全書（KEGG）通路富集分析，隨后進行最小絕對值收斂和選擇算子（LASSO）回歸分析和支持向量機（SVM）分析，得到柴胡疏肝散治療抑郁癥的潛在靶標，并進行ROC生存分析和分子對接驗證以及基于斑馬魚抑郁模型的實驗驗證。結果 共從柴胡疏肝散水提物中鑒定126個成分，“成分-抑郁癥”交集靶點共114個，度值≥11（中位數(shù)）的潛在藥效物質44個，排名前5的成分為柴胡皂苷 A、川陳皮素、異甘草素、洋川芎內酯 A 和柚皮苷查爾酮。獲得 5 個關鍵基因 FOS、TNF、NF-κB1、CXCR2 和IDO1，作為柴胡疏肝散抗抑郁癥的潛在靶標。分子對接結果顯示，5個關鍵成分與 5個潛在靶標均具有良好的結合能，其中與 NF-κB1和 TNF結合能最佳。通過體內實驗研究，在確定柴胡疏肝散抗抑郁有效的前提下，柴胡疏肝散可顯著降低斑馬魚抑郁模型體內促炎因子白細胞介素6（IL-6）和腫瘤壞死因子-α（TNF-α）的表達水平（P＜0.001）。在基因水平上，柴胡疏肝散可顯著降低斑馬魚抑郁模型 NF-κB1、TNF-α和 IL-6 mRNA的表達水平（P＜0.05）。結論 柴胡疏肝散可通過柴胡皂苷A、川陳皮素等多種成分調控TNF-α/NFκB信號通路發(fā)揮抗抑郁作用。

Objective To explore the pharmacodynamic substances and mechanism of Chaihu Shugan Powder (CHSGP) in the treatment of depression and to verify the experimental results. Methods The chemical constituents of CHSGP were analyzed by UPLC-Q-TOF-MS/MS. Traditional Chinese Medicine System Pharmacology Database (TCMSP) and SwisTargetPrediction database were used to obtain and screen the component target. Differential gene analysis and weighted gene co-expression network analysis (WGCNA) analysis were performed on GSE19738 gene chip data using Limma package and hclust function, and depression related targets were obtained and intersected with component targets. The chemical composition-target network diagram was drawn, the degree value was calculated, and the pharmacodynamic substances were screened. The intersection genes were concentrated by GO and KEGG, followed by LASSO regression analysis and SVM analysis, to obtain the potential targets of CHSGP in the treatment of depression, and ROC survival analysis and molecular docking verification as well as experimental verification based on zebrafish depression model. Results A total of 126 chemical constituents were identified from CHSGP water extracts, 114 intersections of "chemical-depression" were identified, and 44 potential pharmacodynamic substances with a degree ≥ 11 (median) were identified. The top 5 components were saikosaponin A, nobiletin, 2',4,4'-trihydroxy-chalcone, ligustrolide A and naringin chalcone. Five key genes, FOS, TNF, NF- κB1, CXCR2 and IDO1, were obtained as potential targets for GHSGP antidepression. Molecular docking results showed that the five key components had good binding energy with five potential targets, and the binding energy with NF-κB1 and TNF was the best. In vivo studies, under the premise of determining the efficacy of CHSGP as an antidepressant, CHSGP can significantly reduce the expression levels of pro-inflammatory factors IL-6 and TNF-α in zebrafish depression models (P <0.001). At the gene level, CHSGP could significantly reduce the mRNA expression levels of NF-κB1, TNF-α and IL-6 in zebrafish depression model (P <0.05). Conclusion CHSGP can regulate the TNF-α/NF-κB signaling pathway through a variety of components such as saikosaponin A and chenorin to play an antidepressant role.

R285.5

黑龍江省重點研發(fā)計劃指導類項目（GZ20210110）；黑龍江省中醫(yī)藥管理局中醫(yī)藥經典普及化專項課題項目（ZYW2022-063）；黑龍江省自然科學基金項目（LH2022H001）；哈爾濱商業(yè)大學產業(yè)化項目支持計劃（XL0079）