A Preliminary Study On The Interaction Between Hepatitis E Virus And Filamin A

Hepatitis E Virus(HEV)is a viral hepatitis infection that is spread by the fecal-oral route and poses a major health risk to humans.Each year,roughly 20.1million people are infected worldwide,with 70,000 people dying.The link between HEV and the host,on the other,is not fully understood.Filament protein A(FLNa)is an actin binding protein that facilitates viral entrance by binding the HIV-1 receptor to the actin cytoskeleton.Influenza A virus(IAV)by interacting with FLNa disrupts the host FLNa-TRAF2 relationship,activating the JNK pathway,which promotes IAV replication.However,no research on the link between FLNa and HEV have been published.The interaction between HEV and FLNa was investigated at the clinical patient,animal model,and cell levels in this work,and the following conclusions were reached:(1).HEV infection inhibited FLNa expression(1)q RT-PCR was used to detect FLNa expression of the serum of HEV RNA-positive patients and healthy people.The results revealed that FLNa expression of HEV-infected patients serum was suppressed.(2)A correlation study revealed that HEV inhibited FLNa expression.(3)FLNa expression was considerably suppressed in the liver tissues of hepatocellular carcinoma patients infected with HEV,according to immunohistochemical data.(4)FLNa expression was detected using q RT-PCR in BALB/c mice and theses monkeys infected with HEV.HEV infection decreased FLNa expression,according to the findings.(5)Immunohistochemistry,q RT-PCR and western blot were used to detect FLNa expression in Hep G2 cells infected with HEV.The findings revealed that HEV infection inhibited the expression of FLNa.(2).Interaction between HEV and FLNa(1)A confocal cell assay revealed that HEV could co-locaize with FLNa.(2)Western blot analysis revealed that the HEV Hel domain could have a role in FLNa regulation.(3)The HEV Hel domain was found to be involved in control of FLNa in Co-IP and confocal cell assays.(4)The FLNa knockdown plasmid was produced successfully,and the knockdown was confirmed using q RT-PCR and western blot.(5)Using cell immunofluorescence and q RT-PCR,researchers discovered that knocking down FLNa promotes virus multiplication,and immunofluorescence data revealed that HEV infection inhibits FLNa expression.(3).HEV regulates NF-κB signaling pathway by inhibiting FLNa expression(1)After HEV infection and FLNa knockdown,RT-PCR was utilized to detect the expression of interferon stimulating genes(ISGs).Both HEV infection and FLNa knockdown resulted in aberrant expression of several interferon boosting genes,according to the findings.(2)q RT-PCR was used to detect the expression of the NF-κB gene.The results showed that following infection and FLNa knockdown in HEV,the NF-κB gene was considerably suppressed.(3)The inhibitory impact on the NF-κB inhibitor QNZ was detected by Western blot,and the NF-κB signaling a pathway was significantly decreased.Both HEV infection and FLNa knockdown were found to severely suppress the NF-κB signaling pathway.(4)A fluorescence test was used to look at the expression of apoptosis and autophagy following HEV infection and FLNa knockdown.Both HEV infection and FLNa knockdown were observed to cause enhanced inflammatory responses such apoptosis and autophagy.(5)The ubiquitination level of proteins in HEV infection and FLNa knockdown was determined using western blot and immunohistochemistry analysis.The findings revealed that ubiquitination occurred in HEV infection and FLNa knockdown cells,and that the level of ubiquitination was suppressed.