目的 探究柴胡疏肝散治療抑郁癥的藥效物質(zhì)及作用機(jī)制并進(jìn)行實(shí)驗(yàn)驗(yàn)證。方法 應(yīng)用超高效液相色譜-四極桿飛行時(shí)間質(zhì)譜聯(lián)用（UPLC-Q-TOF-MS/MS）技術(shù)對(duì)柴胡疏肝散化學(xué)成分進(jìn)行分析；采用中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫(kù)及分析平臺(tái)（TCMSP）和SwisTargetPrediction數(shù)據(jù)庫(kù)獲取并篩選成分靶點(diǎn)；運(yùn)用Limma包和hclust函數(shù)對(duì)GSE19738基因芯片數(shù)據(jù)進(jìn)行差異基因分析和加權(quán)基因共表達(dá)網(wǎng)絡(luò)（WGCNA）分析，獲取抑郁癥相關(guān)靶點(diǎn)，并與成分靶點(diǎn)取交集。繪制成分-靶點(diǎn)網(wǎng)絡(luò)圖，計(jì)算度值，篩選藥效物質(zhì)。交集基因通過(guò)基因本體（GO）功能富集分析和京都基因與基因組百科全書(shū)（KEGG）通路富集分析，隨后進(jìn)行最小絕對(duì)值收斂和選擇算子（LASSO）回歸分析和支持向量機(jī)（SVM）分析，得到柴胡疏肝散治療抑郁癥的潛在靶標(biāo)，并進(jìn)行ROC生存分析和分子對(duì)接驗(yàn)證以及基于斑馬魚(yú)抑郁模型的實(shí)驗(yàn)驗(yàn)證。結(jié)果 共從柴胡疏肝散水提物中鑒定126個(gè)成分，“成分-抑郁癥”交集靶點(diǎn)共114個(gè)，度值≥11（中位數(shù)）的潛在藥效物質(zhì)44個(gè)，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(nèi)酯 A 和柚皮苷查爾酮。獲得 5 個(gè)關(guān)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(biāo)。分子對(duì)接結(jié)果顯示，5個(gè)關(guān)鍵成分與 5個(gè)潛在靶標(biāo)均具有良好的結(jié)合能，其中與 NF-κB1和 TNF結(jié)合能最佳。通過(guò)體內(nèi)實(shí)驗(yàn)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(yú)抑郁模型體內(nèi)促炎因子白細(xì)胞介素6（IL-6）和腫瘤壞死因子-α（TNF-α）的表達(dá)水平（P＜0.001）。在基因水平上，柴胡疏肝散可顯著降低斑馬魚(yú)抑郁模型 NF-κB1、TNF-α和 IL-6 mRNA的表達(dá)水平（P＜0.05）。結(jié)論 柴胡疏肝散可通過(guò)柴胡皂苷A、川陳皮素等多種成分調(diào)控TNF-α/NFκB信號(hào)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(diǎn)研發(fā)計(jì)劃指導(dǎo)類(lèi)項(xiàng)目（GZ20210110）；黑龍江省中醫(yī)藥管理局中醫(yī)藥經(jīng)典普及化專(zhuān)項(xiàng)課題項(xiàng)目（ZYW2022-063）；黑龍江省自然科學(xué)基金項(xiàng)目（LH2022H001）；哈爾濱商業(yè)大學(xué)產(chǎn)業(yè)化項(xiàng)目支持計(jì)劃（XL0079）