| PrefaceInfluenza is the acute virus air tube disease which is originated by influenza virus o Influenza is the very important infection diseases for human.The disease incidences reach the first place of all the infection diseases.So far,the most effect ways to prevent influenza is by influenza vaccine.To anticipate the periodicity of influenza for sure is very difficult.So it brings many difficult for the production of influenza vaccine.Then it is the most important thing to invest the material of anti-influenza virus.Methionine Enkephalin(MEK) is the natural substance secreted by adrenal gland and drived from pre hormone.It has been reported that MEK takes part in immune regulation and exerts broad affects on both immune system and endocrine system.Investigation indicated that MEK can markedly increase both cellular and humeral immunity on the rats infected with influenza B virus.MEK in combination with other antivirus drug can also prolong the survival of rats with virus pneumonia.In order to look at insight and effects on influenza B virus in vitro and in vivo by MEK and provide data to the development of MEK as potential vaccine we performed this study.Materials and methods一,Materials1.Agents and Drugs:MEK:Product made in Penta-biotech co..USA,Batch No.63231,MW:574.2.Influenza virus:Influenza B virus(titers 1:64):A strain preserved and passed by Liaoning CDC.3.Chicken embryos:Chicken embryo of 8-11 days were provided by company of breeding in Liaoning.Chaoyang city.4.Medin-Darby Canine Kidney(MDCK):The MDCK monolayer cells with 17-18 passages are from Liaoning CDC and grow every well for 24 hours before use.二,Methods1.Applying MEK on MDCK infected with influenza B virus:Dividing the experiment into 3 groups:1,normal control group,using the virus maintaining liquid,2, virus control group,Putting in 100TCID50 influenza B virus,3 testing group,Putting MEK with 10-5,10-6,10-7,10-8,10-9,10-10,10-11,10-12,10-13,10-14mol/L and 100 TCID50 influenza B viruses in different orders in MDCK cell cultivating wells,at 34℃5%CO2,incubate for 72 hours and look at the CPE under microscope,let it stand at -70℃and test HA titers.2.Applying MEK on chick embryo infected with influenza B virus:Dividing the experiment into 3 groups:1,normal control group,using 0.9%NS,2,virus control group,Putting in 100EID50 influenza B virus,3 testing group,Putting MEK with 10-12mol/L and 100EID50 influenza B virus in different orders in chick embryo,at 34℃5%CO2,incubate for 72 hours,let it stand at -20℃for 1 hour and test HA titers.Results1.Applying MEK on MDCK infected with influenza B virusThere is no CPE observed on normal control group while virus control and testing group appears equally cell lesion.In the HA titer experiment,the normal control group gives negative HA,however the virus control and testing group give positive HA titer 1:64.2.Applying MEK on chick embryo infected with influenza B virusIn the HA titer experiment,the normal control group gives negative HA,however the virus control give positive HA titer 1:64,the group which add MEK first then incubate for 24 hours and add influenza B viruses later,give positive 6.8±1.93,has the statistical compare with the virus control group.DiscussionsMEK is the important regulatory substance in nervous system,endocrine system and immune system.Investigations prove there are MEK receptors in lymphocytes, macrophage and natural killer cells.The suitable dosage of MEK can activate the immune cells through binding with the receptors. Through the studying data we got on influenza B virus by MEK we know that, MEK does not have the direct effect on virus in vitro,but can inhibit growth of influenza B virus in vivo.This means that MEK have indirect effect in vivo.Through the immune system to secret cytokines such asγ-IFN,interleukin or other functional proteins to inhibit or block virus life or replication etc.Further study to reveal concrete mechanisms is still on process.Our study provides a meaningful mode of action to the MEK as non specific,preventive or protective vaccine for human being in the future.Conclusions1.There is no any direct anti virus effect for MEK while working on MDCK against influenza B virus.2.MEK can exert anti virus activity against influenza B virus in chicken embryo model. |
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