Preliminary Study On Mechanism Of Cork Water Extract Against Herpes Simplex Virus

Objective Herpes simplex virus(HSV)is the earliest and most common herpes virus found in humans.According to the difference in antigenicity,it can be divided into herpes simplex virus Ⅰ(HSV-1)and herpes simplex virus Ⅱ(HSV-2).HSV-1 often causes facial and oral skin and mucous membrane infections,such as herpes labialis,keratitis,etc.In addition,studies have shown that Alzheimer’s disease and HIV(human immunodeficiency virus)transmission are related to HSV-1 infection.There is currently no effective vaccine worldwide.Commonly used therapeutic medications,such as acyclovir,famciclovir,and ganciclovir,have a certain effect on reducing clinical symptoms,but only for the complete eradication of herpes simplex virus infection.It is believed that the effects of these medications are minimal.Besides,due to the widespread use of traditional antiviral medications,the potential drug-resistant strains has increased.In view of the above reasons,it is urgent to develop new anti-HSV-1 drugs.In recent years,studies have confirmed that Cork inhibits HSV-1,but the mechanism is not clear.This study investigated the anti-herpes simplex virus type Ⅰ activity of water extract of Cork by in vitro experiments,preliminary explored the mechanism of action Cork extract against herpes simplex virus type Ⅰ,and provided a new therapeutic strategies and medicine targets for diseases caused by herpes simplex virus type Ⅰ infection.Methods1.He La cells were used to amplify HSV-1 virus,and the virus virulence was identified using Median Tissue Culture Infectious Dose(TCID50).2.The Tetramethylazozolium salt(MTT)colorimetric method was adopted detect the toxicity of Cork extract to Hela cells.3.He La cells were inoculated with HSV-1 after a dose-dependent Cork extract stimulation.The virus proliferation was detected by fluorescence emission of the virus through fluorescence microscope.4.Q-PCR was performed to detect the transcriptional expression of the surface proteins g B and g D of the virus capsule membrane.5.Changes in the transcription level of viral nuclear transcription factor κB(NF-κB)were detected by Q-PCR,and Western-blot and immunofluorescence were performed to detect the protein expression NF-κB,in order to explore the antiviral action mechanism of cork water extract.Results1.Virulence was identified after virus amplification,TCID50 = 10-6.7.2.The MTT assay of the inhibitory rate of Cork extract on Hela cells showed that medication concentration at 2,10,and 50 ng / ml did not affect Hela cell proliferation.3.Fluorescence expression of the virus was observed by fluorescence microscope.This result suggested that the virus proliferation was significantly reduced by adding 10 and 50 ng / ml.4.The Q-PCR results proved that at medication concentrations of 10 and 50 ng / ml,the expression level of g D on the viral envelope membrane protein decreased,but g B transcription level increased.5.The transcription factor NF-κB transcription level increased at 10 and 50 ng / ml,but the expression level of NF-κB protein in western-blot and immunofluorescence showed a downward trend.Conclusion1.The water extract of Phellodendron amurense has anti-HSV-1 effects in vitro.2.The antiviral mechanism of the water extract of Phellodendron chinensis may be related to the inhibition of the transcription of viral envelope protein g D.3.Cork water extract can effectively regulate the activation of NF-κB to inhibit virus replication.