Research On The Mechanism Of Hsp90 Inhibitors Inhibit The Entry Of HSV-1 Infects Nerve Cells By Promoting The Polymerization Of Microfilaments

Research background: Herpes simplex virus type Ⅰ(HSV-1)is an enveloped virus with double-stranded DNA,belonging to the alpha subfamily of the herpesvirus family,as a common human pathogenic virus that can cause herpesvirus encephalitis.At present,the commonly used anti-HSV-1 drugs in clinic are mainly nucleoside drugs represented by acyclovir,the treatment drugs are single and the resistance is becoming more and more serious.Therefore,it is urgent to find new antiviral drug targets.Hsp90 plays an important role in the life cycle of HSV-1,and Hsp90 inhibitors have significant antiviral activity.Therefore,this research mainly focuses on the antiviral efficacy and mechanism of Hsp90 inhibitors in HSV-1 infected nerve cells.To provide a theoretical basis and experimental basis for Hsp90 as a new anti-HSV-1 drug target.Methods and Results: In this study,the effects of two Hsp90 inhibitors,17 AAG and AT533,on the titer of HSV-1 were detected by virus titer experiments,and inhibition of the Hsp90 inhibitors on the proliferation of HSV-1/F strains and three ACV resistant virus strains were detected through q RT-PCR experiments.The effects of Hsp90 inhibitors and si RNA knockdown of Hsp90α and Hsp90β on the entry stage of HSV-1 infected neurons were investigated by Western blot,q RT-PCR and plaque reduction experiments,then it was verified on the primary nerve cells.Finally,the function and molecular mechanism of Hsp90 in the entry stage of HSV-1 infected nerve cells were explored through immunofluorescence,flow cytometry and Western blot experiments.The experimental results show that Hsp90 inhibitors have obvious antiviral effects on both HSV-1/F strains and ACV-resistant virus strains.Hsp90 inhibitors inhibit the entry of HSV-1 infected neurons,specifically,Hsp90 Inhibitors promoted the attachment of HSV-1 and inhibited the penetration of HSV-1.Hsp90 inhibitors caused microfilament polymerization.Hsp90 inhibitors caused the abnormal localization of microfilament depolymerization factor Cofilin.Conclusion and Significance: Hsp90 plays an important role in the life cycle of HSV-1 infected nerve cells.Hsp90 inhibitors can inhibit HSV-1 infection of nerve cells and have antiviral effects on ACV resistant virus strains,and have a good antiviral effect on ACV resistant strains.Hsp90 inhibitor may be caused by abnormal intracellular localization of microfilament depolymerization factor Cofilin,leading to microfilament polymerization,which is conducive to virus attachment,but excessive polymerization of microfilament hinders the penetration of HSV-1,eventually,Hsp90 inhibitors inhibit HSV-1 from entering nerve cells.This study explored the antiviral efficacy and mechanism of Hsp90 inhibitors in nerve cells,and provided a theoretical basis for Hsp90 as a new anti-HSV-1 drug target point.