Study On Antiviral Activity And Mechanism Of Three Main Active Ingredients From Zedoary Turmeric Oil Against H1N1

Highly pathogenic influenza viruses pose a serious public health threat to humans.Although vaccines are available,antivirals are needed to efficiently control disease progression and virus transmission due to the emergence of drug-resistant viral strains.In this study,curcumol,curdione,germacrone are the three main active ingredients from Zedoary Turmeric oil.We found that they could inhibit the replication of H1N1 influenza virus effectively.Based on the comparison and analysis of structural active sites of three compounds,we try to find the possible target and related pathways to explain the antiviral mechanism in vitro or vivo,so as to provide a new evidence for influenza treatment.In vitro,we tested the drug effects on cell proliferation by CCK-8kit,and found that there was non-toxic effect on cells in a certain range of drug concentration.By using DNA electrophoresis,semi-quantitative PCR,real time PCR,and report gene assay,we detected whether the compounds had broad spectrum antiviral effects.The results show that in the 5 kinds of virus,drug shave a stabilizing effect on H1N1 virus in a dose-dependent manner.The drugs were estimated to play an effect on the entry and replication stage of the virus detected by real time PCR.In the tests of cell immunofluorescence and virus titer,we found that these compounds could inhibit the virus NP protein expression and decrease virion generation in a time-dependent manner.Moreover,the drugs have a protective effect in vivo by reducing the number of virus titer bothin serum and blood cells and alleviating lung tissue lesions.Especially in the early stage of infection,germacrone show the most obvious protective effect on mice.Furthermore,the technology of gene expression microarray and quantitative protein spectrum analysis were applied to screen the differential genes,and proteins to find the possible mechanism which was interfered by germacrone.The methods of real time PCR and WB were used to verify the expression of the differential genes and proteins.Three compounds,curcumol,curdione,and germacrone belonging to sesquiterpenoids,can effectively fight against influenza virus,among which germacrone has the best effect.These compounds can affect the virus' s stage of entry and replication,inhibit the expression of viral NP protein and reduce virus virion.Mass spectrum data analysis show that germacron can significantly down-regulate TAP1 which is induced by virus to negatively regulate the antiviral innate immune response.The results of the study preliminarily reveal the mechanism of the anti-H1N1 influenza virus.Germacrone provides some theoretical support for the development of clinical application of influenza virus.