Based on network pharmacology and molecular docking, explore the mechanism of Lianhua Qingwen capsule’s blood components intervening in cytokine storm to prevent and treat novel coronavirus pneumonia
Novel coronavirus disease 2019 (COVID-19) is caused by coronavirus infection. Its main clinical characteristics are fever, fatigue and dry cough. Some patients quickly become severe one week after the onset of the disease. Severe cases can further develop into acute respiratory distress syndrome (ARDS), septic shock, metabolic acidosis that is difficult to correct, and multiple organ failure (MODS). It is worth noting that a small number of early-stage mild patients may suddenly worsen in the later stage, leading to multiple organ failure and death, mainly due to the triggering of cytokine storms.
Cytokine storm, also known as inflammation storm and hypercytoplasmic syndrome, is mainly caused by the rapid and massive release of various cytokines after immune system dysfunction, leading to extensive edema of lung tissue and acute lung injury. It is closely related to the poor prognosis of COVID-19 and is an important turning point in disease progression, ultimately developing into ARDS, MODS, and even death. It can be seen that anti-inflammatory treatment may be an effective method to curb cytokine storm and prevent COVID-19.
Lianhua Qingwen Capsule (hereinafter referred to as “Lianhua Qingwen Capsule”), as the representative of traditional Chinese patent medicines and simple preparations for public health emergencies of respiratory infectious diseases, is mainly made from Yinqiao Powder and Maxi Ganshi Decoction, including 13 traditional Chinese medicines such as honeysuckle, forsythia suspensa, rhubarb, etc., which has the effect of clearing away the plague, detoxifying, purging the lung and relieving fever. Previous studies have confirmed that Lianhua Qingwen can significantly improve symptoms such as fever, cough, sputum production, shortness of breath, and fatigue in patients by regulating the expression of inflammatory cytokines, leading to a significant decrease in the rate of weight loss. This is of great significance for the treatment of COVID-19. This study will be based on the blood components of Lianhua Qingwen, and explore the possible mechanism of intervention in cytokine storm prevention and treatment of COVID-19 through network pharmacology and molecular docking, in order to provide theoretical guidance for clinical application.

This study identified 17 effective blood entering components of Lianhua Qingwen through literature review, and used network topology analysis to determine the core components of emodin, formononetin, rutin, gallic acid, and liquiritigenin; Obtained 47 potential targets for the intervention of cytokines storm in the prevention and treatment of COVID-19 using the blood components of Lianhua Qingwen, including 22 core targets such as AKT1, IL-6, TP53, JUN, TNF, CASP3, IL1B, EGF, etc. In order to better interpret the functions involved in the targets, this study conducted GO enrichment analysis and KEGG pathway analysis on 47 targets. The results showed that the mechanism of action of the blood components of Lianhua Qingwen mainly enriched RNA polymerase II to initiate positive regulation of transcription from RNA polymerase II promoter, inflammatory response, positive regulation of NF – κ B transcription factor activity, and other inflammation related biological processes; KEGG pathway analysis mainly involves pathways in cancer, TNF signaling pathway, MAPK signaling pathway, PI3K Akt signaling pathway, HIF-1 signaling pathway, NF – κ B signaling pathway, etc. Molecular docking showed that emodin, formononetin, rutin, gallic acid, and liquiritigenin compounds in the blood components of Lianhua Qingwen have good binding ability to AKT1, IL-6, TP53, JUN, and TNF core targets. It is speculated that Lianhua Qingwen’s blood components may regulate relevant targets, inhibit the secretion of inflammatory cytokines, block the binding of SARS-CoV-2 virus and ACE2 protein, prevent SARS-CoV-2 from infecting human cells, and thus exert the effect of intervening in cytokine storms.
In order to further analyze whether the components of Lianhua Qingwen entering the bloodstream have a regulatory effect on multi organ tissue injury and immune injury, multiple organ tissue injury and immune injury related targets were retrieved from the database, and the intersection was taken with the targets of Lianhua Qingwen entering the bloodstream. The results showed that the proportion of intersecting targets of Lianhua Qingwen capsules entering the bloodstream with multi organ tissue injury and immune injury was 1.6% to 2%. Due to the high homology between SARS virus and coronavirus, the intersection between the blood components of Lianhua Qingwen and SARS virus was taken, and the ratio of the intersection target between Lianhua Qingwen and SARS virus was found to be 4.3%. The blood components of Lianhua Qingwen have the potential to regulate immunity, protect tissues and organs, and resist viruses. The KEGG pathway analysis results showed that multi organ tissue damage, immune damage, and SARS mainly involve cancer pathways, PI3K Akt, MAPK, TNF, HIF-1, and other signaling pathways. Therefore, based on target prediction and KEGG pathway analysis, this study will explore the intervention of Lianhua Qingwen’s blood components in cytokine storm prevention and treatment of COVID-19 from three aspects: clearing antigens, regulating the immune system, and protecting tissues and organs.
Clearing the source of resistance – antiviral. Novel coronavirus is the pathogen of COVID-19. The State Key Laboratory of Respiratory Diseases, the First Affiliated Hospital of Guangzhou Medical University found that Lianhua Qingwen could significantly inhibit the activity of novel coronavirus in vitro, reduce the virus content in cell membrane and cytoplasm, and inhibit the over activation of cytokines. Research has found that ACE2 is a receptor for SARS-CoV-2 host cells, and the binding of SARS-CoV-2 to ACE2 is an important link in viral infection of human cells. After SARS-CoV-2 infection, it enters alveolar cells and replicates a large amount of cytokines in a short period of time, ultimately forming a cytokine storm that triggers inflammation. Ho et al. found that emodin inhibits SARS-CoV infection by blocking the binding of SARS-CoV protein to ACE2 receptors, thereby exerting antiviral effects. Previous studies have shown that when rutin is 200mg/kg, the inhibition rate of lung index in influenza mice is 27.3%, indicating a significant anti influenza virus effect. The inhibitory effect on influenza virus may occur during the release and diffusion stages. MAPK is a known major cellular signaling pathway that can be activated by various viruses. Previous reports on human coronavirus infection have shown that the p38MAPK pathway is activated, and its downstream regulatory protein phosphorylation is enhanced, promoting the production of pro-inflammatory cytokines and leading to cytokine storms. Wang et al. found through in vivo and in vitro experiments that rhein can inhibit the adsorption and replication of influenza A virus (IAV), possibly through the inhibition of MAPK and NF – κ B signaling pathways. It has been confirmed that glycyrrhetinic acid, the main component of licorice, can inhibit the replication of SARS CoV virus in Vero cells cultured in vitro, and also inhibit the adsorption and permeation of the virus in the early stages of the replication cycle. The above indicates that the blood components of Lianhua Qingwen have broad-spectrum antiviral effects, which may further suppress cytokine storms from the source by inhibiting virus reproduction.
Immune regulation – resisting inflammatory storms. Immune disorders are the core of cytokine storms. After the virus enters the body, it disrupts the immune regulatory network, leading to an imbalance between pro-inflammatory cytokines and anti-inflammatory cytokines, causing the body to enter a highly inflammatory state and triggering cytokine storms. Through the Venn diagram, it was found that the components of Lianhua Qingwen entering the bloodstream have 133 common targets with immune damage, indicating that the components of Lianhua Qingwen entering the bloodstream may inhibit inflammatory storms through immune regulation. According to existing research, peripheral blood CD4+and CD8+T lymphocytes in COVID-19 patients are significantly reduced, but the Th17 subset of CD4+T lymphocytes is increased, and CD8+T lymphocytes contain higher concentrations of cytotoxic particles, indicating that to some extent, the patient’s immune system is severely damaged. IL-6 is an inflammatory cytokine. The secretion of IL-6 helps to evaluate the severity of COVID-19, and the dynamic change of IL-6 level can be used as an indicator for monitoring the condition of patients with severe COVID-19. TNF – α is mainly a small molecule protein secreted by mononuclear macrophages, which releases inflammatory factors to participate in the inflammatory response, thereby exacerbating the occurrence and development of diseases. Previous studies have found that activating the NF – κ B pathway leads to overexpression of pro-inflammatory cytokines such as TNF – α and IL-6. Blocking the NF – κ B signaling pathway not only inhibits virus transmission, but also suppresses the development of related inflammation. Glycyrrhetinic acid can play an immune regulatory role in sheep Mycoplasma pneumoniae (MO) infection by increasing macrophage proliferation after MO infection. Emodin, as one of the main components of rhubarb, can not only inhibit early inflammatory cytokines, but also suppress the inflammatory cascade reaction and reduce the expression of “secondary” inflammatory cytokines. By inhibiting NF – κ B p65 phosphorylation, it can reduce the expression of IL-6 and TNF α in peripheral blood, inhibit LPS induced macrophage cytokine release, and have ideal anti-inflammatory effects. Previous studies have shown that mangiferin can reduce the expression of inflammatory factors in brain tissue, mainly by controlling the activation of the SphK1/SIP signaling pathway, thereby inhibiting the NF – κ B signaling pathway and further improving the damage of inflammatory factors to brain tissue. Liu et al. observed that salidroside can significantly reduce pro-inflammatory cytokines and lung inflammation in serum, significantly inhibit pro-inflammatory cytokines such as TNF – α, IL-1 β, and IL-6, and significantly increase the level of anti-inflammatory cytokine IL-10. The above indicates that the blood components of Lianhua Qingwen inhibit the NF – κ B signaling pathway, downregulate the expression of pro-inflammatory cytokines, upregulate the levels of anti-inflammatory cytokines, and balance between pro-inflammatory cytokines and anti-inflammatory cytokines, thereby intervening in cytokine storms.
Organ and tissue protection. The consequence of cytokine storm is multiple organ failure, where excessive cytokine release can lead to tissue damage and, in severe cases, death. In clinical biochemical testing indicators, it was found that COVID-19 mainly damages the lungs, followed by varying degrees of damage to the heart, liver, and kidneys. According to the Venn diagram created based on the components of Lianhua Qingwen entering the bloodstream and the targets of various organ injuries, it can be seen that Lianhua Qingwen entering the bloodstream has intersecting targets for various organ injuries, which may alleviate tissue and organ damage and enhance the body’s immune function through these targets. PI3K belongs to intracellular phosphatidylinositol kinase, and Akt is a very important downstream active target of PI3K, belonging to a serine/threonine protein kinase. The PI3K/Akt signaling pathway is upregulated in LPS induced chronic liver injury, significantly affecting the NF – κ B signaling pathway and leading to transcriptional changes in cells, ultimately resulting in liver dysfunction; Studies have shown that amygdalin reduces the expression of PI3K and phosphorylated AKT in cells, indicating that amygdalin enhances anti-inflammatory ability and has a significant protective effect against liver injury by inhibiting the PI3K/AKT and NF – κ B signaling pathways. Hu et al. found that LPS induced inflammation of TNF – α, IL-1, and IL-6 is a key factor in the deterioration of acute lung injury (ALI). Therefore, inhibiting the excessive production and expression of pro-inflammatory cytokines to reduce LPS induced ALI cannot be ignored. Emodin plays a protective role in severe acute pancreatitis (SAP) lung injury rat model by downregulating the inflammatory expression of NF – κ B, TNF – α, and IL-1 β, thus protecting SAP lung injury and the body. The components of Lianhua Qingwen entering the bloodstream have the effect of reducing tissue and organ damage by inhibiting the PI3K/AKT and NF – κ B inflammatory signaling pathways.
In summary, exploratory studies on the components of Lianhua Qingwen entering the bloodstream through network pharmacology and molecular docking have shown that Lianhua Qingwen entering the bloodstream has the characteristics of multi-component, multi-target, and multi pathway. It can clear antigens, regulate immunity, and protect tissues and organs by regulating relevant cytokines and signaling pathways. This study provides a material basis and theoretical basis for the effective prevention and treatment of COVID-19 with Lianhua Qingwen Capsules.