Study On The Mechanism Of Sulforaphane Up-regulation Of P53 Protein To Inhibit The Replication Of Enterovirus EV-D68

Background:Enterovirus EV-D68 belongs to the genus Enterovirus of the small RNA virus family.It replicates rapidly and mainly causes respiratory symptoms in newborns and infants.Mild symptoms are mainly similar to fatigue and cough caused by influenza,and severe symptoms include acute flaconic myelitis(AFM).So far,there is no specific drug or vaccine,so the research of anti-EV-D68 virus drug is particularly important.The first is to determine whether enterovirus EV-D68 infects cells and causes cytopathic changes,injuries or even death.Secondly,we used EV-D68 virus structural protein VP1 as the detection index to search for traditional Chinese medicine monomer compounds that could inhibit viral replication.Then,after determining the ability of sulforaphane monomer compound to inhibit the replication of enterovirus EV-D68,the possible antiviral mechanism of sulforaphane was analyzed.Finally,the upregulation of p53 protein was determined to be an important factor in the viral ability of sulforaphane,and the mechanism of inhibiting enterovirus EV-D68 by sulforaphane was further analyzed.Method:RD cells(human striated muscle cells),He La cells(human cervical epithelium He La cells)and H1299 cells(human non-small cell lung cancer cells)were studied.Firstly,morphological observation and flow cytometry were used to determine the effect of enterovirus EV-D68 on host cells.Secondly,using VP1,the structural protein of EV-D68 virus,as the detection index,Western Blot and other experiments were conducted to search for traditional Chinese medicine monomer compounds that could inhibit the replication of viral protein.After finding a method to inhibit the replication of enterovirus EV-D68,According to the antiviral mode of other drugs,the antiviral mode and pathway of the traditional Chinese medicine monomer compound were screened.Finally,taking this pathway and target as the object,q PCR and TCID50 experiments were used to further analyze the inhibitory effect and mechanism of the TCM monomer compound on the replication of enterovirus EV-D68.Results & Conclusions:1.Sulforaphane can alleviate the morphological damage caused by enterovirus EV-D68,while reducing virus replication at the protein level,genome level,and virus particle count.2.p53 protein can inhibit viral replication of enterovirus EV-D68.3.Sulforaphane can upregulate the content of p53 protein and inhibit the replication of enterovirus EV-D68.When the cell does not contain p53 gene,Sulforaphane cannot play an antiviral role..Innovations:1.Sulforaphane can inhibit the replication of enterovirus EV-D68 virus.2.Overexpression of P53 protein can play a role in inhibiting enterovirus EV-D68.3.Sulforaphane can inhibit the replication of enterovirus EV-D68 by upregulating the content of p53 protein.