背景:H1N1是引起季节性流感的甲型流感病毒(IAV)的主要亚型之一,严重威胁人类健康。临床上使用一种称为清兴颗粒(QX)的中药组合来治疗流行性流感。然而,它的化学成分很复杂,潜在的药理机制仍然未知。
方法:根据两个标准从TCMSP数据库中收集了QX的有效成分:药物相似度(DL≥0.18)和口服生物利用度(OB≥30%)。SwissADME用于预测有效成分的潜在目标,和Cytoscape用于为QX创建“Herb-Component-Target”网络。此外,与H1N1相关的目标是从GeneCards数据库中收集的,OMIM,和GEO。从KEGG检索与自噬相关的靶标,HAMdb,和HADb数据库。QX的交叉点目标,甲型H1N1流感,和自噬使用维恩图进行鉴定。之后,使用利用数据库STRING构建的Cytoscape蛋白-蛋白相互作用网络筛选关键靶标。通过GO分析和KEGG富集分析观察重叠靶标的生物学功能和信号通路。采用高效液相色谱法(HPLC)测定QX的主要化学成分,其次是分子对接。最后,通过动物实验验证了QX治疗H1N1的作用机制。
结果:共鉴定出786个潜在的QX靶标和91个有效组分。有5420个与H1N1相关的靶标和821个与自噬相关的靶标。QX所有目标的交点,H1N1和自噬产生了75个交叉靶标。最终,选择了10个核心目标:BCL2,CASS3,NFKB1,MTOR,JUN,TNF,HSP90AA1,EGFR,HIF1A,还有MAPK3通过HPLC鉴定QX的主要化学成分,在195min内分离出7种标记成分,是苦杏仁苷,葛根素,黄芩苷,连翘苷,沃戈诺赛德,黄芩素,和Wogonin.分子对接结果表明,BCL2、CASP3、NFKB1和MTOR能很好地与化合物结合。在动物研究中,QX通过上调p-mTOR/mTOR和p62的表达和下调LC3的表达来降低H1N1感染小鼠肺组织的退行性改变,从而抑制自噬。
结论:根据本研究的网络药理学分析和实验确认,QX可能通过调节自噬来治疗H1N1感染,降低LC3的表达,增加p62和p-mTOR/mTOR的表达。
BACKGROUND: H1N1 is one of the major subtypes of influenza A virus (IAV) that causes seasonal influenza, posing a serious threat to human health. A traditional Chinese medicine combination called Qingxing granules (QX) is utilized clinically to treat epidemic influenza. However, its chemical components are complex, and the potential pharmacological mechanisms are still unknown.
METHODS: QX\'s effective components were gathered from the TCMSP database based on two criteria: drug-likeness (DL ≥ 0.18) and oral bioavailability (OB ≥ 30%). SwissADME was used to predict potential targets of effective components, and Cytoscape was used to create a \"Herb-Component-Target\" network for QX. In addition, targets associated with H1N1 were gathered from the databases GeneCards, OMIM, and GEO. Targets associated with autophagy were retrieved from the KEGG, HAMdb, and HADb databases. Intersection targets for QX, H1N1 influenza, and autophagy were identified using Venn diagrams. Afterward, key targets were screened using Cytoscape\'s protein-protein interaction networks built using the database STRING. Biological functions and signaling pathways of overlapping targets were observed through GO analysis and KEGG enrichment analysis. The main chemical components of QX were determined by high-performance liquid chromatography (
HPLC), followed by molecular docking. Finally, the mechanism of QX in treating H1N1 was validated through animal experiments.
RESULTS: A total of 786 potential targets and 91 effective components of QX were identified. There were 5420 targets related to H1N1 and 821 autophagy-related targets. The intersection of all targets of QX, H1N1, and autophagy yielded 75 intersecting targets. Ultimately, 10 core targets were selected: BCL2, CASP3, NFKB1, MTOR, JUN, TNF, HSP90AA1, EGFR, HIF1A, and MAPK3. Identification of the main chemical components of QX by
HPLC resulted in the separation of seven marker ingredients within 195 min, which are amygdalin, puerarin, baicalin, phillyrin, wogonoside, baicalein, and wogonin. Molecular docking results showed that BCL2, CASP3, NFKB1, and MTOR could bind well with the compounds. In animal studies, QX reduced the degenerative alterations in the lung tissue of H1N1-infected mice by upregulating the expression of p-mTOR/mTOR and p62 and downregulating the expression of LC3, which inhibited autophagy.
CONCLUSIONS: According to this study\'s network pharmacology analysis and experimental confirmation, QX may be able to treat H1N1 infection by regulating autophagy, lowering the expression of LC3, and increasing the expression of p62 and p-mTOR/mTOR.