Exploring the active ingredients and pharmacological mechanisms of the oral intake formula Huoxiang Suling Shuanghua Decoction on influenza virus type A based on network pharmacology and experimental exploration

Abstract

ObjectiveTo investigate the active ingredients, underlying anti-influenza virus effects, and mechanisms of Huoxiang Suling Shuanghua Decoction (HSSD).Materials and methodsThe therapeutic effect of HSSD were confirmed through the survival rate experiment of H1N1-infected mice. Then, the HSSD solution and the ingredients absorbed into the blood after treatment with HSSD in rats were identified by UPLC/Q-TOF MS, while the main contents of ingredients were detected by high performance liquid chromatography (HPLC). Next, a systems pharmacology approach incorporating target prediction, gene ontology (GO) enrichment, kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and molecular docking were performed to screen out the active compounds and critical pathways of HSSD in treating influenza. According to prediction results, real-time quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry assay were used to detect the mRNA and protein expression levels of critical targets in H1N1-infected mice lungs.ResultsHuoxiang Suling Shuanghua Decoction improved the survival rate of H1N1-infected mice and prolonged the mice’s lifespan. Besides, HSSD exerts an antivirus effect by decreasing the levels of hemagglutinin (HA) and nucleoprotein (NP) to inhibit the replication and proliferation of H1N1, reducing the lung pathological state, inhibiting the cell apoptosis in the lung, and regulating the abnormal responses of peripheral blood, including GRA, LYM, white blood cell (WBC), PLT, and hemoglobin (HGB). Then, 87 compounds in the HSSD solution and 20 ingredients absorbed into the blood after treatment with HSSD were identified. Based on this, combined with the network analysis and previous research on antivirus, 16 compounds were screened out as the active components. Moreover, 16 potential targets were predicted by network pharmacology analysis. Next, molecular docking results showed stable binding modes between compounds and targets. Furthermore, experimental validation results indicated that HSSD regulates the contents of Immunoglobulin A (IgA), Immunoglobulin M (IgM), and Immunoglobulin G (IgG) in serum, modulating the levels of IFN-γ, IL-6, IL-10, MCP-1, MIP-1α, and IP-10 in the lung tissue, and significantly decreasing the mRNA and protein expressions of TLR4, CD14, MyD88, NF-κB p65, HIF1 α, VEGF, IL17A, and IL6 in the lung tissue.ConclusionHuoxiang Suling Shuanghua Decoction exerts an anti-influenza effect by affecting the expressions of mRNA and protein including TLR4, CD14, MyD88, NF-kB p65, HIF-1α, VEGF, IL17A, IL6, and inhibiting the accumulation of inflammation. Our study provided experimental pieces of evidence about the practical application of HSSD in treating influenza.

Boosting the immunogenicity of the CoronaVac SARS-CoV-2 inactivated vaccine with Huoxiang Suling Shuanghua Decoction: a randomized, double-blind, placebo-controlled study

“…Several herbal tonics used in TCM have shown to enhance immunity with little or no toxicity or side effects, and may potentially be used as vaccine adjuvants to promote the secretion of immune-activating factors, activate B cells and T cells, and enhance the body’s humoral and cellular immunity ( 25 ). Our previous research showed that HSSD may aid the treatment of influenza A virus infection by eliciting anti-inflammatory and immunoregulatory effects ( 18 ). Our proof-of-concept study in mice immunized with SARS-CoV-2 S-RBD recombinant protein showed that HSSD supplementation significantly enhanced serum anti-RBD IgG titers, increased the percentage of CD4 + T cells and boosted the secretion of related cytokines, and regulated adaptive immunity.…”

“…In our previously study, four compounds of HSSD were identified by the HPLC profile, including chlorogenic acid, caffeic acid, scutellarin, and rosmarinic acid, and the contents of two signature ingredients in the HSSD were determined by HPLC method as chlorogenic acid (0.63 mg/g), and hesperidin (3.13 mg/g), respectively. More details on the ingredients of HSSD were published previously ( 19 ). Product standard code of HSSD was GB/T 29602, and its Production License Number was No.…”

“…Metabolomics uses modern detection techniques combined with bioinformatics analysis methods to perform semi-quantitative or quantitative detection of low molecular weight metabolites in blood, urine, feces, and other biological samples after exposure to pathophysiological stimuli, environmental changes, or drug interventions ( 17 ). In a preclinical study, we recently demonstrated that Huoxiang Suling Shuanghua Decoction (HSSD) can treat influenza A virus infection by preventing inflammation and improving immunity ( 18 ). Therefore, before conducting the present study, we tested whether HSSD supplementation was able to boost specific serum IgG antibody titers in mice immunized with recombinant SARS-CoV-2 S-RBD protein.…”