key: cord-0994055-cnx1attk authors: Li, Yuan; Chu, Fuhao; Li, Ping; Johnson, Nadia; Li, Tao; Wang, Yan; An, Rongxian; Wu, Dantong; Chen, Jiena; Su, Zeqi; Gu, Xiaohong; Ding, Xia title: Potential effect of Maxing Shigan decoction against coronavirus disease 2019 (COVID-19) revealed by network pharmacology and experimental verification date: 2021-01-26 journal: J Ethnopharmacol DOI: 10.1016/j.jep.2021.113854 sha: 954553e025b10b4f60a85bde5ce8a0d1f8e6bb4a doc_id: 994055 cord_uid: cnx1attk ETHNOPHARMACOLOGICAL RELEVANCE: Since the occurrence of coronavirus disease 2019 (COVID-19) in Wuhan, China in December 2019, COVID-19 has been quickly spreading out to other provinces and countries. Considering that traditional Chinese medicine (TCM) played an important role during outbreak of SARS and H1N1, finding potential alternative approaches for COVID-19 treatment is necessary before vaccines are developed. According to previous studies, Maxing Shigan decoction (MXSGD) present a prominent antivirus effect and is often used to treat pulmonary diseases. Furthermore, we collected 115 open prescriptions for COVID-19 therapy from the National Health Commission, State Administration of TCM and other organizations, MXSGD was identified as the key formula. However, the underlying molecular mechanism of MXSGD against COVID-19 is still unknown. AIM OF THE STUDY: The present study aimed to evaluate the therapeutic mechanism of MXSGD against COVID-19 by network pharmacology and in vitro experiment verification, and screen the potential components which could bind to key targets of COVID-19 via molecular docking method. MATERIALS AND METHODS: Multiple open-source databases related to TCM or compounds were employed to screen active ingredients and potential targets of MXSGD. Network pharmacology analysis methods were used to initially predict the antivirus and anti-inflammatory effects of MXSGD against COVID-19. IL-6 induced rat lung epithelial type Ⅱ cells (RLE-6TN) damage was established to explore the anti-inflammatory damage activity of MXSGD. After MXSGD intervention, the expression level of related proteins and their phosphorylation in the IL-6 mediated JAK-STAT signaling pathway were detected by western blot. Molecular docking technique was used to further identify the potential substances which could bind to three key targets (ACE2, Mpro and RdRp) of COVID-19. RESULTS: In this study, 105 active ingredients and 1025 candidate targets were selected for MXSGD, 83 overlapping targets related to MXSGD and COVID-19 were identified, and the protein-protein interaction (PPI) network of MXSGD against COVID-19 was constructed. According to the results of biological enrichment analysis, 63 significant KEGG pathways were enriched, and most of them were related to signal transduction, immune system and virus infection. Furthermore, according the relationship between signal pathways, we confirmed MXSGD could effectively inhibit IL-6 mediated JAK-STAT signal pathway related protein expression level, decreased the protein expression levels of p-JAK2, p-STAT3, Bax and Caspase 3, and increased the protein expression level of Bcl-2, thereby inhibiting RLE-6TN cells damage. In addition, according to the LibDock scores screening results, the components with strong potential affinity (Top 10) with ACE2, Mpro and RdRp are mainly from glycyrrhiza uralensis (Chinese name: Gancao) and semen armeniacae amarum (Chinese name: Kuxingren). Among them, amygdalin was selected as the optimal candidate component bind to all three key targets, and euchrenone, glycyrrhizin, and glycyrol also exhibited superior affinity interactions with ACE2, Mpro and RdRp, respectively. CONCLUSION: This work explained the positive characteristics of multi-component, multi-target, and multi-approach intervention with MXSGD in combating COVID-19, and preliminary revealed the antiviral and anti-inflammatory pharmacodynamic substances and mechanism of MXSGD, which might provide insights into the vital role of TCM in the prevention and treatment of COVID-19. profuse expectoration. Shigao is effective in febrile diseases due to exogenous 34 affection with high fever and dire thirst, asthma and cough caused by heat in the lung. 35 Gancao mainly used to modulate the pharmaceutical activity of different herbal 36 medicines, and it can also moisten lung and relieve cough. Four Chinese medicines 37 cooperate with each other to play the main effect of ventilating lung, and relieving 38 cough and asthma. Above all, it is reasonable to believe that MXSGD is the core 39 combination of most clinically effective formulations and has application value. 40 However, the mechanism of MXSGD in the treatment of COVID-19 is still unclear. Therefore, in this study we used network pharmacology to screen active ingredients 59 and potential targets of MXSGD, and to reveal its potential medicinal substance and 60 mechanism in antivirus and anti-inflammatory. Molecular docking was used to further 61 identify the potential substances which could bind to three key targets of COVID-19 62 (ACE2, Mpro and RdRp). Table 305 1, according to the LibDock scores screening results, the components with strong In order to reveal the interaction between the candidate compounds and COVID-19 318 related targets, the binding sites of the compounds and targets were further analyzed. According to the LibDock scores of the compounds and targets in Supplementary 8, 320 amygdalin was selected as the optimal candidate component for ACE2, Mpro, and 321 RdRp. The binding atoms, distance, and interaction category of amygdalin with 322 COVID-19 related targets are presented in Table 3 . 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