Objective To determine the anti-rotavirus(RV)effect of piceatannol(PIC)in vitro and related mechanism.Methods The RV-Wa and RV-SA11 strains were used to infect the MA 104 cell model,and MTT was used to assess the anti-RV adsorption,direct RV inhibition,and anti-RV biosynthesis effects of piceatannol.TCID50 was used to detect the change of virus titer before and after piceatannol treatment.Molecular docking was used to predict the degree of binding of piceatannol to the VP8*protein of the RV-Wa strain.The expression of VP6 mRNA was detected by RT-qPCR.Indirect immunoassay was used to detect the expression of VP6 protein.The expression of VP6,IκBα,NF-κBp65 and p-NF-κBp65 proteins was detected by Western blot.The levels of IL-1p,IL-6,and TNF-α in the cell supernatants were evaluated by ELISA.Results Piceatannol had a dual effect of direct RV inhibition and anti-RV biosynthesis,without anti-RV adsorption.The direct inhibition of RV by piceatannol was studied,which showed that the virus titer decreased greatly after piceatannol treatment.The molecular docking revealed that piceatannol and RV-Wa VP8*protein had a better binding capacity.The anti-RV biosynthesis effect of piceatannol was studied.RT-qPCR showed that piceatannol down-regulated the expression of VP6 mRNA.Western blot showed that RV infection treated with piceatannol down-regulated the expression of VP6 and p-NF-κBp65 proteins,and up-regulate the expression of IκBα protein.ELISA showed that piceatannol reduced the levels of IL-1β,IL-6,and TNF-α in the cell supernatants.Conclusion Piceatannol has a dual effect of direct RV inhibition and anti-RV biosynthesis.The anti-RV effect may be achieved by limiting RV invasion into cells by association with the RV-Wa VP8*protein and lowering the phosphorylation of NF-κBp65,which attenuates the inflammatory responses generated by RV infection.
万方数据
维普
