ABSTRACT
Objective: The objective of this study was to characterize the chemical compounds of a Hanshi-Yufei formulation (HSYF;a modified formulation of a traditional Chinese medicine used for treating COVID-19) to elucidate the mechanism of action and to evaluate potential anti-inflammatory effects of HSYF. Materials and Methods: The chemical constituents of HSYF extract were characterized using UPLC-Q-TOF/MS. Subsequently, a set of TCM network pharmacology methods was applied to identify disease-associated genes and to predict target profiles and pharmacological actions associated with the constituents of HSYF. Then, the antiviral effects of HSYF on H1N1 were assessed in RAW264.7 cells using MTT assays. Expression levels of pro-inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-α following infection of RAW264.7 cells with H1N1 were measured using an enzyme-linked immune sorbent assay (ELISA), and expression levels of inflammatory-related factors were detected using western blotting. Results: In total, 165 chemical constituents (including glycosides, tannins, volatile oils, amino acids, triterpenoids, polyphenols, phenylpropanoids, sesquiterpenes, alkaloids, and flavonoids, among others) were tentatively identified in HSYF. Network pharmacology demonstrated that HSYF can regulate immunomodulatory- and anti-inflammatory-related targets of multiple pathways through its active ingredients, suggesting potential anti-COVID-19 effects. Furthermore, cell viability assays and ELISA showed that HSYF significantly inhibited H1N1 replication in RAW64.7 cells and markedly reduced expression of pro-inflammatory cytokines TNF-α and IL-6 at the proteins level. Conclusions: The results of the present study help improve our understanding of the therapeutic effects of HSYF in COVID-19 treatment from multi-level perspectives.
ABSTRACT
To analyse the differential indicators of COVID-19 in severe and mild cases and to study the factors affecting the immune function of patients and the time required for oropharyngeal swabs to become negative. Age, albumin (ALB) levels, prealbumin (PAB) levels, high-sensitivity C-reactive protein (hs-CRP) levels, platelet counts, lymphocyte counts, neutrophil counts, CD3+, CD4+, CD8+ T cell counts and the time for oropharyngeal swabs to become negative were collected from 37 patients with COVID-19;the neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) were calculated as indicators of inflammation. An independent-sample t test was used to analyse differences between the severe and mild groups, and factors affecting the CD3+, CD4+ and CD8+ T cell counts and the time for the nucleic acid tests of oropharyngeal to convert to negative were identified by single-factor and multifactor analyses. Lymphocyte, ALB, PAB, CD3+, CD4+ and CD8+ T cell levels in the severe group were lower than those in the mild group, the P values were 0.048, 0.004, 0.033, 0.033, 0.015 and 0.013, respectively. The neutrophil count and PLR were higher in the severe group compared with that in the patients of mild group;the P values were all 0.000. Single-factor analysis showed that age, ALB level, PAB level, hs-CRP level, platelet count, the NLR, the PLR and the time to a negative nucleic acid test were the main factors influencing CD3+ T cells;the P values were 0.001, 0.031, 0.001, 0.010, 0.005, 0.002, 0.000 and 0.048, respectively. Age, ALB level, PAB level, hs-CRP level, platelet count, the NLR, the PLR and time to a negative nucleic acid test were the main factors influencing CD8+ T cells;the P values were 0.000, 0.012, 0.000, 0.005, 0.002, 0.004, 0.005 and 0.003, respectively. Age, PAB level, hs-CRP level, platelet count, the NLR and the PLR were the main factors influencing CD4+ T cells;the P values were 0.001, 0.006, 0.030, 0.041, 0.005 and 0.001, respectively. Age, ALB level, PAB level, hs-CRP level, platelet count, the NLR, CD3+ T cell count and CD8+ T cell count were the main factors influencing the time to a negative nucleic acid test in oropharyngeal swabs, and the P values were 0.032, 0.043, 0.013, 0.016, 0.042, 0.049, 0.048 and 0.003, respectively. Multivariate analysis showed that the PLR and platelet count were the main factors influencing CD3+ T cells. The P values were all 0.000. The PLR and platelet count were the main factors influencing CD4+ T cells. The P values were 0.000 and 0.001, respectively. The PLR and platelet count were also the main factors influencing CD8+ T cells. The P values were 0.004 and 0.001. CD8+ T cells affected the time to a negative nucleic acid test in oropharyngeal swab samples, and the P value was 0.002. There were differences in the PLR, PAB level, ALB level and T cells between the severe and mild groups. The platelet count and PLR were the main factors influencing the immune function of patients with COVID-19, and CD8+ T cells influenced the negative conversion time of the nucleic acid test in oropharyngeal swabs.