Lin Lili, An Li, Chen Hui, Feng Lu, Lu Mengjiang, Liu Yuling, Chu Chu, Shan Jinjun, Xie Tong, Wang Xiaorong, Wang Shouchuan
Jiangsu Key Laboratory of Pediatric Respiratory Disease, Institute of Pediatrics, Medical Metabolomics Center, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China.
Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, Nanjing, China.
Front Pharmacol. 2021 Dec 1;12:777689. doi: 10.3389/fphar.2021.777689. eCollection 2021.
Respiratory syncytial virus (RSV) can cause varying degrees of lung inflammation in children. Qingfei Oral Liquid (QF) is effective in treating childhood RSV-induced lung inflammation (RSV-LI) in clinics, but its pharmacological profiles and mechanisms remain unclear. This study combined network Pharmacology, lipidomics, pharmacodynamics, and pathway validation to evaluate the therapeutic mechanisms of QF. Using Cytoscape (v3.8.2) and enrichment analyses from the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO), a global view of the putative compound-target-pathway network was created. The corresponding lipidomic profiles were then used to detect differently activated lipids, revealing the metabolic pathway, using ultra-high-performance liquid chromatography linked to hybrid Quadrupole-Exactive Orbitrap mass spectrometry (UHPLC-Q-Exactive Orbitrap MS). Meanwhile, the efficiency of QF, the enrichment pathway, and the excessive autophagy inhibition mechanisms were validated in RSV-infected mice models. The network pharmacology results demonstrated 117 active compounds acted directly upon 101 core targets of QF against RSV-LI. The most significantly enriched pathway was the PI3K/Akt/mTOR signaling pathway ( < 0.05). In addition, untargeted lipidomics were performed, and it was revealed that higher lung levels of DAG 30:0, DAG 30:5, DAG 32:0, DAG 16:0_18:0, DAG 17:0_17:0, DAG 34:1, DAG 36:0, DAG 36:1 in the RSV-LI group were decreased after QF administration ( < 0.05, FC > 1.2). Lipin-1, a key enzyme in DAG synthesis, was increased in the RSV-LI mouse model. Animal experiments further validated that QF inhibited the PI3K/Akt/mTOR signaling pathway, with lower lung levels of phosphorylated PI3K, AKT and mTOR, as well as its related proteins of lipin-1 and VPS34 ( < 0.01). Finally, pharmacodynamic investigations indicated that QF reduced airway inflammation caused by excessive autophagy by decreasing lung levels of RSV F and G proteins, Beclin-1, Atg5, and LC3B II, IL-1 and TNF-α ( < 0.05). Lipidomic-based network pharmacology, along with experimental validation, may be effective approaches for illustrating the therapeutic mechanism of QF in the treatment of RSV-LI.
呼吸道合胞病毒（RSV）可导致儿童出现不同程度的肺部炎症。清肺口服液（QF）在临床上对治疗儿童RSV引起的肺部炎症（RSV-LI）有效，但其药理特性和作用机制尚不清楚。本研究结合网络药理学、脂质组学、药效学和通路验证来评估QF的治疗机制。使用Cytoscape（v3.8.2）以及来自京都基因与基因组百科全书（KEGG）和基因本体论（GO）的富集分析，构建了假定的化合物-靶点-通路网络的全局视图。然后使用相应的脂质组学图谱来检测不同激活的脂质，通过超高效液相色谱与混合四极杆-高分辨质谱联用（UHPLC-Q-Exactive Orbitrap MS）揭示代谢途径。同时，在RSV感染的小鼠模型中验证了QF的疗效、富集通路和过度自噬抑制机制。网络药理学结果表明，117种活性化合物直接作用于QF针对RSV-LI的101个核心靶点。最显著富集的通路是PI3K/Akt/mTOR信号通路（<0.05）。此外，进行了非靶向脂质组学分析，结果显示，QF给药后，RSV-LI组中肺组织中DAG 30:0、DAG 30:5、DAG 32:0、DAG 16:0_18:0、DAG 17:0_17:0、DAG 34:1、DAG 36:0、DAG 36:1的水平降低（<0.05，FC>1.2）。DAG合成中的关键酶Lipin-1在RSV-LI小鼠模型中增加。动物实验进一步验证了QF抑制PI3K/Akt/mTOR信号通路，肺组织中磷酸化PI3K、AKT和mTOR以及其相关蛋白Lipin-1和VPS34的水平较低（<0.01）。最后，药效学研究表明，QF通过降低肺组织中RSV F和G蛋白、Beclin-1、Atg5和LC3B II、IL-1和TNF-α的水平，减轻了过度自噬引起的气道炎症（<0.05）。基于脂质组学的网络药理学以及实验验证可能是阐明QF治疗RSV-LI治疗机制的有效方法。
