Investigation of the Mechanism of Action of JinHuangJieDu against SARS-CoV-2 Infection and Inflammation Based on Network Pharmacology and Experimental Validation
Listed in
This article is not in any list yet, why not save it to one of your lists.Abstract
Objective
Since the emergence and widespread of coronavirus disease (COVID-19) in 2019, traditional Chinese medicine (TCM) and its formulations have been extensively utilized in China for the prevention and treatment of COVID-19, demonstrating significant clinical efficacy. However, there is an ongoing need for novel TCM preparations. The aim of this study was to assess the potential and mechanism of a de novo designed TCM preparation: JinHuangJieDu (JHJD), which is composed of “ JIN YIN HUA ”, “ HUANG QIN ”, “ GUANG HUO XIANG ”, “ BAI ZHI ”, “ PU GONG YING ” and “ BO HE ”, for the treatment of COVID-19 using network pharmacology analysis and in vitro experimental validations.
Methods
First, enzyme linked immunosorbent assay (ELISA) was established to validate whether JHJD inhibits the binding of wild type and mutant receptor binding domain (RBD) of spike protein to angiotensin-converting enzyme 2 (ACE2). Inhibition of SARS-CoV-2 infection by JHJD was assessed with pseudovirus neutralization assay using SARS-CoV-2 pseudovirus and COS7-hACE2 cells. Network pharmacology analysis using Encyclopedia of Chinese Traditional Medicine (ETCM) and Traditional Chinese Medicine Systems Pharmacology databases (TCMSP) were further performed to identify the meridional network, active ingredients and targeted genes of JHJD. Interaction networks were constructed and gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis were performed using Cytoscape (3.7.1) software with ClueGO and CluePedia plugins. Vital genes were identified using CytoHubba and MCODE plugins. In vitro experiments were performed to confirm one of the targeted crucial gene and to validate whether JHJD may mitigate inflammation caused by SARS-CoV-2.
Results
JHJD effectively blocked the binding of both wild type and Omicron RBD of SARS-CoV-2 to human ACE2 and completely inhibited the infection of SARS-CoV-2 pseudovirus to ACE2-positive host cells in vitro. Network pharmacology analysis uncovered that JHJD comprises a total of 378 active ingredients and 339 targeted genes, with its primary actions likely focused on lung, stomach and spleen. A total of 63 significant KEGG pathways were enriched, predominantly related to viral infection and inflammation pathways. Two pivotal genes, CCND1 and HIF1A , were identified by intersecting the MCODE module genes with different node algorithms in CytoHubba. In vitro validation confirmed that JHJD could target HIF-1α, reducing virus-induced inflammation, which may potentially mitigate the symptoms of COVID-19.
Conclusion
The present study demonstrated that a de novo designed TCM JHJD could effectively inhibit the infection of SARS-CoV-2 through blocking of the interaction between RBD and ACE2. The inhibitory effect appears to be particularly potent against Omicron variants, as JHJD demonstrated enhanced efficacy in inhibiting the RBD-ACE2 interaction for several Omicron variants. Furthermore, JHJD could mitigate the virus-induced inflammation by targeting HIF-1α. Collectively, this study suggested that JHJD preparation holds promise as a potential therapeutic candidate for COVID-19, as it not only inhibits the viral entry into host cells but also reduces the inflammatory responses triggered by the infection.
