Network pharmacology and experimental validation of Maxing Shigan decoction in the treatment of influenza virus-inducedferroptosis

Influenza is an acute viral respiratory infection that has caused high morbidity and mortality worldwide.Influenza A virus(IAV)has been found to activate multiple programmed cell death pathways,including ferroptosis.Ferroptosis is a novel form of programmed cell death in which the accumulation of intracellular iron promotes lipid peroxidation,leading to cell death.However,little is known about how influenza viruses induce ferroptosis in the host cells.In this study,based on network pharmacology,we predicted the mechanism of action of Maxing Shigan decoction(MXSGD)in IAV-induced ferroptosis,and found that this process was related to biological processes,cellular components,molecular function and multiple signaling pathways,where the hypoxia inducible factor-1(HIF-1)signaling pathway plays a significant role.Subsequently,we constructed the mouse lung epithelial(MLE-12)cell model by IAV-infected in vitro cell experiments,and revealed that IAV infection induced cellular ferroptosis that was characterized by mitochondrial damage,increased reactive oxygen species(ROS)release,increased total iron and iron ion contents,decreased expression of ferroptosis marker gene recombinant glutathione peroxidase 4(GPX4),increased expression of acyl-CoA synthetase long chain family member 4(ACSL4),and enhanced activation of hypoxia inducible factor-1α(HIF-1α),induced nitric oxide synthase(iNOS)and vascular endothelial growth factor(VEGF)in the HIF-1 signaling pathway.Treatment with MXSGD effectively reduced intracellular viral load,while reducing ROS,total iron and ferrous ion contents,repairing mitochondrial results and inhibiting the expression of cellular ferroptosis and the HIF-1 signaling pathway.Finally,based on animal experiments,it was found that MXSGD effectively alleviated pulmonary congestion,edema and inflammation in IAV-infected mice,and inhibited the expression of ferroptosis-related protein and the HIF-1 signaling pathway in lung tissues.

Network pharmacology research of mechanism of Maxing Shigan Decoction in treating COVID-19

Background:The COVID-19 has a huge negative impact on people’s health.Traditional Chinese Medicine(TCM)has a good effect on viral pneumonia.It is of great practical significance to study its pharmacology.Methods:The ingredients and targets of each herb in Maxing Shigan Decoction which obtained from Traditional Chinese Medicine Systems Pharmacology(TCMSP)database,and the related targets of COVID-19 were screened by GeneCards database based on the network pharmacology.Venn was used to analyze the intersection target between active ingredients and diseases.Cytoscape software was used to construct an active ingredient-disease target network.The protein-protein interaction network was constructed by STRING database and Cytohubba was used to screen out the key targets.Gene Ontology(GO)functional enrichment analysis and KEGG pathway analysis were performed by DAVID database.Results:In this study,a total of 134 active ingredients and 229 related targets,198 targets of COVID-19 and 48 common targets of drug-disease were chosen.Enrichment items and pathways were obtained through GO and KEGG pathway analysis.The predicted active ingredients were quercetin,kaempferol,luteolin,naringenin,glycyrol,and the key targets involved IL6,MAPK3,MAPK8,CASP3,IL10,etc.The results showed that the active ingredients of Maxing Shigan Decoction acted on multiple targets which played roles in the treatment of COVID-19 by regulating inflammation,immune system and other pathways.Conclusions:The main contribution of this paper is to use data to mine the principles of the treatment of COVID-19 from the pharmacology of these prescriptions,and the results can be provided theoretical reference for medical workers.

Different Effects of Mahuang Decoction and Maxing Shigan Decoction on Animal Temperature Tropism and Correlation to Differences of Cold and Hot Nature of Chinese Materia Medica

Objective To establish an objective method for evaluating the intrinsic characteristics between cold and hot nature of Chinese materia medica (CMM) through the different effects of Mahuang decoction (MHD) and Maxing Shigan decoction (MSD) on animal temperature tropism. Methods The equipment with cold/hot pads was used to investigate the variety of the temperature tropism between two groups of mice treated by MHD and MSD, respectively. Meanwhile, the activities of adenosine triphosphatase (ATPase), superoxide dismutase, succinate dehydrogenase, and malondialdehyde were measured. Results After treated by MHD, the macroscopic behavioral index of remaining rate on warm pad (40 ℃ ) of mice decreased significantly (P < 0.05), suggesting the enhancement of cold tropism, meanwhile, the internal indices of ATPase activity and oxygen consuming volume increased significantly (P < 0.05), suggesting the enhancement of energy metabolism. On the other hand, the above-mentioned indices in MSD group changed on the inverse way. Conclusion The relative drug nature of MHD and MSD revealed in this study is consistent with the theoretical prognostication or definition. It indicates that the internal cold and hot nature of CMM could be reflected in ethological way on the changes of animal temperature tropism which might be internally regulated by body energy metabolism.