School of Pharmacy, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, 550025, China.
College of Life Science, Guizhou University, Guiyang, Guizhou, 550025, China.
J Ethnopharmacol. 2021 Jan 10;264:113364. doi: 10.1016/j.jep.2020.113364. Epub 2020 Sep 9.
Lonicerae japonicae flos (LJF, the dried flower bud or newly bloomed flower of Lonicera japonica Thunb.), a typical herbal medicine, targets the lung, heart and stomach meridian with the function of clearing heat and detoxication. It ameliorated inflammatory responses and protected against acute lung inflammation in animal models. Acute lung injury (ALI) is a kind of inflammatory disease in which alveolar cells are damaged. However, a network pharmacology study to thoroughly investigate the mechanisms preventing ALI has not been performed.
In this study, we examined the main active ingredients in LJF and the protective effects of LJF on LPS-induced ALI in rats.
First, the main active ingredients of LJF were screened in the TCMSP database, and the ALI-associated targets were collected from the GeneCards database. Then, we used compound-target and target-pathway networks to uncover the preventive mechanisms of LJF. Furthermore, we assessed the preventive effects of LJF in an LPS-induced rat model with the RNA-Seq technique to validate the possible molecular mechanisms of the effects of LJF in the treatment of ALI.
The network pharmacology results identified 28 main active compounds in LJF, and eight chemical components highly related to the potential targets, which were potential active compounds in LJF. In all, 94 potential targets were recognized, including IL6, TNF, PTGS2, APP, F2, and GRM5. The pathways revealed that the possible targets of LJF involved in the regulation of the IL-17 signalling pathway. Then, in vivo experiments indicated that LJF decreased the levels of proinflammatory cytokines (TNF-, IL-1, and IL-6) in serum and bronchoalveolar lavage fluid, decreased the levels of oxidative stress factors (MDA and MPO) and increased the activities of SOD and GSH-Px in lung tissue. The RNA-Seq results revealed that 7811, 775 and 3654 differentially expressed genes (DEGs) in Ctrl (control group), ALI-LJF (Lonicerae japonicae flos group) and ALI-DXSM (dexamethasone group), respectively. KEGG pathway analysis showed that the DEGs associated with immune response and inflammation signalling pathways and the IL-17 signalling pathway were significantly enriched in LJF. Compared with those in ALI, the expression of CXCL2, CXCL1, CXCL6, NFKBIA, IFNG, IL6, IL17A, IL17F, IL17C, MMP9 and TNFAIP3, which are involved in the IL-17 signalling pathway, were significantly decreased in the LJF group according to the qRT-PCR analyses.
In view of the network pharmacology and RNA-Seq results, the study identified the main active ingredient and potential targets of LJF involved in protecting against ALI, which suggests directions for further research on LJF.
金银花（Lonicerae japonicae flos，忍冬科植物忍冬的干燥花蕾或初开的花）是一种典型的草药，归肺、心、胃经，具有清热解毒的功效。它在动物模型中改善了炎症反应并保护急性肺炎症。急性肺损伤（ALI）是一种肺泡细胞受损的炎症性疾病。然而，尚未进行全面研究预防 ALI 的网络药理学研究。
本研究旨在探讨金银花的主要活性成分及其对 LPS 诱导的大鼠 ALI 的保护作用。
首先，在 TCMSP 数据库中筛选金银花的主要活性成分，并从 GeneCards 数据库中收集与 ALI 相关的靶点。然后，我们使用化合物-靶点和靶点-通路网络来揭示金银花的预防机制。此外，我们使用 RNA-Seq 技术评估了 LPS 诱导的大鼠模型中金银花的预防作用，以验证金银花治疗 ALI 的可能分子机制。
网络药理学结果鉴定出金银花中的 28 种主要活性化合物，以及 8 种与潜在靶点高度相关的化学物质，这些都是金银花中的潜在活性化合物。共有 94 个潜在靶点被识别，包括 IL6、TNF、PTGS2、APP、F2 和 GRM5。途径分析表明，金银花可能通过调节 IL-17 信号通路来发挥作用。然后，体内实验表明，金银花降低了血清和支气管肺泡灌洗液中促炎细胞因子（TNF-α、IL-1 和 IL-6）的水平，降低了氧化应激因子（MDA 和 MPO）的水平，并提高了肺组织中 SOD 和 GSH-Px 的活性。RNA-Seq 结果显示，Ctrl（对照组）、ALI-LJF（金银花组）和 ALI-DXSM（地塞米松组）组分别有 7811、775 和 3654 个差异表达基因（DEGs）。KEGG 通路分析表明，与免疫反应和炎症信号通路以及 IL-17 信号通路相关的 DEGs 在金银花中显著富集。与 ALI 相比，根据 qRT-PCR 分析，金银花组 CXCL2、CXCL1、CXCL6、NFKBIA、IFNG、IL6、IL17A、IL17F、IL17C、MMP9 和 TNFAIP3 等参与 IL-17 信号通路的基因表达显著降低。
基于网络药理学和 RNA-Seq 结果，本研究鉴定了金银花防治 ALI 的主要活性成分和潜在靶点，为进一步研究金银花提供了方向。
