Effect And Mechanism Of Curcumol On Encephalomyocarditis Virus

Objective: sixteen traditional Chinese medicine compounds were screened for anti-encephalomyocarditis virus(EMCV)activity in vitro.Then,the mechanism of drug anti-EMCV was studied in vitro,which provided new ideas for clinical treatment of EMCV infection.Methods: The maximal non-toxic concentration(MNTC),cytotoxic concentration 50(CC50),maximal inhibition rate(MIR)and effective concentration 50(EC50)against EMCV were measured using MTT and antiviral assays on baby hamster syrian kidney(BHK-21)cells.On this basis,The drugs with MIR﹥50%and SI﹥3 were screened.Through EMCV load,the anti-viral effect of compounds weres quantitatively determined using real-time quantitative PCR.The effect of compounds on the expression of IFN-β was checked using real-time quantitative PCR and ELISA.Western blot was used to determine the amounts of MDA5,MAVS,TANK,IRF3 and P-IRF3 proteins in HEK-293 T cells infected with EMCV.Results: The MTT assay was used to investigate the cytopathic rate of the compounds to find out its MNTC and CC50 values.With the exception of geniposide and glycyrrhizic acid,all other drugs exhibit an“S” type dose-dependent relationship.The CC50 of the tested compounds were ranged from 27.8 to 4000μg/m L,and their MNTC values ranged from 12.5 to 4000 μg/m L.These results showed that these compounds possessed different cytotoxicity for BHK-21 cells.The results showed that on BHK-21 cells,compared with the positive drug ribavirin,the MIR of curcumol,matrine and baicalin was greater than that of ribavirin,but the SI value was much smaller than the latter.Real-time PCR results showed that curcumol,matrine,and baicalin could significantly reduce viral load(p﹤0.05).The MNTC and CC50 values of curcumol on HEK-293 T cells were 25 and 124 μg/m L,respectively,which were less than the MNTC and CC50 values of ribavirin.Curcumol and ribavirin have MIR﹥50%and SI﹥3.Real-time quantitative PCR results showed that curcumol could significantly reduce viral load(p﹤0.05).The results showed that curcumol had significant anti-EMC activity in different cells.q PCR and ELISA results showed that curcumol can significantly increase the expression of IFN-β in HEK-293 T cells infected with EMCV(p﹤0.05).Western blot results showed that curcumol can promote the expression of MDA5 and inhibit EMCV-mediated TANK protein.Degradation,promote the expression of p-IRF3,while the expression of MAVS and IRF3 remains unchanged.Conclusions: Curcumol,matrine and baicalin have anti-EMC infection in BHK-21 cells in vitro,and curcumol has the best effect.Curcumol can alleviate the degradation of TANK protein by EMCV and increase the expression of p-IRF3,thereby significantly increasing the expression of IFN-β and exerting its antiviral effect.