Study On The Anti-influenza Virus Mechanisms Of Compounds Santin And DMO-CAP

Influenza pandemic caused by influenza viruses causes tens of thousands of deaths every year,and poses a serious threat to social stability.The two main measures to treat influenza are the prevention of influenza vaccine and antiviral drugs.However,due to the different influenza virus strains each year,the effect of influenza vaccine on influenza prevention is not significant.In addition,the high mutagenicity and recombination rate of influenza viruses lead to resistance of anti-influenza drugs targeting the virus.Therefore,it is more and more urgent to develop new anti-influenza drugs.Artemisia rupestirs L.is a herbal plant of Compositae.It mainly distributes in the northern Xinjiang of China.It is a traditional medicine used by ethnic groups to treat common cold.As one of the main active ingredients of Artemisia rupestirs L.,total flavonoids of Artemisia rupestirs L.has good anti-influenza activity.In this study,we determined the anti-influenza activity of 50% ethanol eluent from Artemisia rupestirs L.,and further studied its anti-influenza mechanism.Objective: To determine whether several isolated monomer compounds of 50% ethanol elution from Artemisia rupestirs L.have anti-influenza virus activity in vitro,and to explore the mechanism of action of compounds with anti-influenza virus activity.Methods: Guided by antiviral pharmacology,the antiviral activity of several isolated monomer compounds from 50% ethanol eluent of Artemisia rupestirs L.was screened by CPE screening method.Secondly,Cytopathic effect(CPE)assay,viral titer assay,western blot,real-time Quantitative real-time RT-PCR and Immunofluorescence assay were used to further verify the anti-influenza virus activity of the selected monomer compounds at the cellular and molecular levels.Finally,the anti-influenza mechanism of the compounds was studied by hemagglutinin inhibition assay,neuraminidase inhibition assay,siRNA gene silencing and Luciferase assay.Result: Preliminary screening by CPE showed that Santin and DMO-CAP had better anti-influenza activity in vitro.The anti-influenza effect of the two drugs was further verified at the cellular and molecular level.In the study of the anti-influenza mechanism of Santin and DMO-CAP,we obtained the following results:In vitro evaluation of anti-influenza activity showed that Santin had anti-influenza activity but not broad-spectrum anti-influenza effect,while DMO-CAP had broad-spectrum anti-influenza effect.In the mechanism study,compound Santin inhibits influenza virus replication through regulating MAPKs and NF-?B signaling pathways.Compound DMO-CAP inhibits influenza virus replication by activating by activating heme oxygenase-1-mediated IFN response.Conclusion: The results showed that Santin and DMO-CAP isolated from 50% ethanol elution of Artemisia rupestirs L.have anti-influenza activity in vitro,with different anti-influenza mechanisms.Therefore,our study provides more theoretical basis for the vacancy of the current research on the antiviral mechanism of Artemisia rupestirs L.