通过网络药理学-分子对接的方法，探讨四方木皮抗炎的作用机制，并用斑马鱼炎症模型进行验证。利用网络药理学预测四方木皮抗炎的有效成分、潜在核心靶点和信号通路。用脂多糖(LPS)诱导斑马鱼炎症模型，以细胞凋亡率和活性氧(ROS)生成率等指标来评价四方木皮水提物和 70%乙醇提取物的抗炎活性，用qPCR验证预测的主要靶点。预测发现，四方木皮的潜在抗炎靶点有 121 个，PPI分析显示，四方木皮抗炎主要作用于信号传导和转录激活因子 3(signal transducer and ac-tivator of transcription 3，STAT3)、血管内皮生长因子A(vascular endothelial growth factor A，VEGFA)、表皮生长因子(epidermal growth factor，EGF)、肿瘤坏死因子(tumor necrosis factor，TNF)、肿瘤蛋白p53(tumor protein p53，TP53)、基质金属蛋白酶9(ma-trix metalloprotein 9，MMP9)、c-fos原癌蛋白(c-fos proto-oncogene protein，FOS)、环状素受体 1(estrogen receptor 1，ESR1)、趋化因子 8(c-x-c motif chemokine ligand 8，CXCL8)、白细胞分化抗原 8(cluster of differentiation 8，CD8)等靶点;基因本体(Gene On-tology，GO)分析显示，生物过程主要作用于抑制细胞凋亡过程、基因表达的正调控、细胞增殖的正向调节;京都基因和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes，KEGG)分析表明，MAPK信号通路、PI3K-Akt信号途径、HIF-1信号通路等可能在四方木皮抗炎中起关键作用;分子对接显示，5 种关键成分与核心靶点均具有较强的结合力。斑马鱼动物实验表明，四方木皮可显著抑制炎症导致的ROS形成，降低幼鱼体内细胞死亡，抑制了组织中炎症反应的增强。此外，与空白组相比，模型组中核因子(nuclear factor，NF)-κB、TP53、FOS、衔接蛋白复合物-1(adaptor protein complex-1，AP-1)、丝裂原活化蛋白激酶P38(mitogen-activated protein kinases P38，P38)的转录水平显著上调;与模型组相比，四方木皮水提物和 70%乙醇提取物组的斑马鱼组织中NF-κB、TP53、FOS、AP-1、P38 的mRNA表达明显下调。四方木皮通过多成分、多靶点发挥抗炎作用，且其抗炎作用可能与抑制MAPK信号通路有关。
Anti-inflammatory effect and its mechanism of Saracae Cortex based on zebrafish model and network pharmacology
This paper aims to explore the anti-inflammatory mechanism of Saracae Cortex by using network pharmacology and mole-cular docking methods and verify it through the inflammation model of zebrafish.The effective components,potential core targets,and signaling pathways of Saracae Cortex were obtained by using network pharmacology.A lipopolysaccharide(LPS)-induced inflammation model of zebrafish was established to evaluate the anti-inflammatory activity of aqueous extract and 70% ethanol extract of Saracae Cor-tex with cell apoptosis rate and reactive oxygen species(ROS)production rate as indicators.qPCR was performed to verify the main targets predicted by network pharmacology.The prediction found that there were 121 potential anti-inflammatory targets in Saracae Cor-tex.Protein-protein interaction(PPI)analysis showed that Saracae Cortex mainly acted on signal transducer and activator of transcrip-tion 3(STAT3),vascular endothelial growth factor A(VEGFA),epidermal growth factor(EGF),tumor necrosis factor(TNF),tumor protein p53(TP53),matrix metalloprotein 9(MMP9),c-fos proto-oncogene protein(FOS),estrogen receptor 1(ESR1),c-x-c motif chemokine ligand 8(CXCL8),cluster of differentiation 8(CD8),and other targets.Gene Ontology(GO)analysis showed the biological process mainly acted on the inhibition of apoptosis,the positive regulation of gene expression,and the positive regulation of cell proliferation.Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis showed that the mitogen-activated protein kinase(MAPK)signaling pathway,PI3K-Akt signaling pathway,and hypoxia-inducible factor 1(HIF-1)signaling pathway may play a key role in anti-inflammation of Saracae Cortex.Molecular docking verified that five key compounds had a strong binding force with their corresponding core target.Zebrafish animal experiments showed that Saracae Cortex could significantly inhibit ROS formation and re-duce cell apoptosis in juvenile fish caused by inflammation and inhibit the further enhancement of inflammatory response in tissue.In addition,compared with the blank group,the transcription levels of nuclear factor kappa-B(NF-κB),TP53,FOS,adaptor protein complex-1(AP-1),and mitogen-activated protein kinases P38(P38)were significantly up-regulated in the model group.Compared with the model group,the mRNA expression of NF-κB,TP53,FOS,AP-1,and P38 was significantly down-regulated in zebrafish tis-sue treated with aqueous extract and 70% ethanol extract of Saracae Cortex.Saracae Cortex plays an anti-inflammatory role through multiple components and targets,and its anti-inflammatory effect may be related to the inhibition of the MAPK signaling pathway.
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