The Effect Of Different Treatment Methods On RLH Signaling Pathway In The Mice Infected By Influenza A Virus (FM1 Strain)

ObjectiveTo study the effects of oseltamivir,berberine and three kinds of Chinese herbal medicine,such as new Elsholtzia drink,Gui-ma-ge-ban tang and yingtao powder,on the RLH signaling pathway in the lungs of mice infected with influenza A virus（FM1 strain）.To explore the mechanism of antiviral effect of these drugs,which will provide the experimental basis for anti-influenza virus drugs discovery and development in future.Methods1.To determine viral LD50 after amplifying influenza A virus（FM1 strain）,and establish the RIG-I-/-mice and normal wide-type C57BL/6 mice influenza modes.2.To treat mice with oseltamivir,,monomeric berberine and three kinds of traditional Chinese medicine by gavage.To daily record and observe the general condition,mortality,weight of mice and so on.3.On the seventh day after virus infection,to detect the changes of lung index,analyze the survival rate,and take the pathological in each group of mice.4.To detect the ratio of Th1/Th2,Th17/Treg in mice by flow cytometry.5.To detect the expression of RIG-I,NF-κB and MAVS in RLH signaling pathway of mouse lung immune cells at the gene and protein levels by RT-PCR and Western-blot.Results1.To obtain the suspension of influenza A virus（FM1 strain）by chicken embryo chorioallantoic cavity expansion method.And the LD50 of FM1 for C57BL/6 mice was 2^-2.83/50μl.2.Oseltamivir,berberine and Gui-ma-ge-ban tang could enhance the survival rate of normal wild type C57BL/6 mice infected by FM1,and reduce their lung index by alleviating lung inflammation of normal wild type C57BL/6 mice infected by FM1.3.Influenza virus infection can lead to the increased ratio of Th1/Th2 or Th17/Treg,which enhance inflammatory response and weaken anti-inflammatory function.Oseltamivir berberine and Gui-ma-ge-ban tang could reduce the ratio of Th1/Th2 and Th17/Treg.4.After RLH signaling pathway activated by Influenza virus,oseltamivir and berberine down-regulated the expression of RIG-I,MAVS and NF-κB in RLH signaling pathway.ConclusionAfter mice infected with influenza virus,RLH signal transduction pathway was activated in their lungs immune cells,which would cause lung inflammation response,increase lung index,and up-regulate the Th1/Th2 and Th17/Treg ratio.And Oseltamivir,berberine and Gui-ma-ge-ban tang could effectively reduce the indicators of normal wide-type C57BL/6mice infected with influenza virus.The berberine had good effect on prohibit influenza A virus as oseltamivir,which could regulate RLH signaling pathway of the infection.