In Vitro Antiviral Activity Of Valeriana Jatamansi Jonse Extract Against Rotavirus

Objective:Taking Wa strain of rotavirus as the research object,the antiviral activity of Valeriana jatamansi Jonse extract against rotavirus was studied in vitro,which provided theoretical basis for the application of Valeriana Jatamansi Jonse against rotavirus.Methods:MA104 cells were cultured in vitro to amplify human rotavirus Wa strain.The VP6 gene of rotavirus was selected to construct the standard plasmid.The virus copies number of clinical rotavirus positive stool samples was detected by RT-qPCR with SYBR method to verify the standard plasmid.Different concentrations of Valeriana jatamansi Jonse extract respectively acted on MA004 cells cultured in vitro,then OD value was measured by CCK-8 method after 72 hours of culture,and the cell survival rate was calculated.The titer of virus was detected by TCID50 method The antiviral effect of Valeriana jatamansi Jonse extract against rotavirus was studied by comprehensive antiviral experiments in vitroResults:The standard plasmid of rotavirus VP6 gene was successfully constructed.The standard curve equation was y=-0.2542x+11.547,R2=0.9993,R2>0.99,and the linearity of Ct value was good in the range of 13.67～33.56.The amplification curve was stable and the Melt curve was single.The copies number of clinical rotavirus positive stool samples were as follows:the original sample 574790.0995 copies/ul,the 10-1 dilution sample 46977.73704 copies/ul,and the 10-2 dilution sample 2600.2913 copies/ul.The titer of rotavirus was TCID50=10-5 6/0.1 ml.The maximum non-toxic concentration of Valeriana jatamansi Jonse extract,Patchouli alcohol,DMSO and Ribavirin on MA104 cells were 1.17 mg/ml,15.63 ug/ml and 0.5%and 0.12 mg/ml,respectively.Ribavirin group and Valeriana jatamansi Jonse extract group could block the adsorption of rotavirus,and the difference was statistically significant(P<0.05).Patchouli alcohol group could block the adsorption of rotavirus,but the difference was not statistically significant(P>0.05).Valeriana jatamansi Jonse extract group had stronger effect on blocking the adsorption of rotavirus than Ribavirin group,and the difference was statistically significant(P<0.05).Ribavirin group,Valeriana jatamansi Jonse extract group and Patchouli alcohol group could direct kill rotavirus,but the difference was not statistically significant(P>0.05).Ribavirin group,Valeriana jatamansi Jonse extract group and Patchouli alcohol group all inhibited the proliferation of rotavirus,and the difference was statistically significant(P<0.05).The ihibiting rotavirus proliferation effect of Valeriana jatamansi Jonse extract group was stronger than that of Patchouli alcohol group,and the difference was statistically significant(P<0.05);The ihibiting rotavirus proliferation effect of Ribavirin group was stronger than that of Patchouli alcohol group,and the difference was statistically significant(P<0.05),The ihibiting rotavirus proliferation effect of Valeriana jatamansi Jonse group was stronger than that of Ribavirin group,but the difference was not statistically significant(P>0.05)Conclusion:The standard plasmid was constructed based on VP6 gene of rotavirus,which was high specificity and sensitivity,used for detecting the copies number of rotavirus for absolute quantification.Valeriana jatamansi Jonse extract could block the adsorption of rotavirus and inhibit proliferation of rotavirus,Patchouli alcohol could inhibit proliferation of rotavirus.