Preliminary Study On Antiviral Effects Of Plant Polyphenols Polysaccharides In Vitro

Objective: Plant polyphenols and polysaccharides have anti-oxidation,anti-tumor,antibacterial,anti-virus and scavenging free radical and other biological functions.Antiviral experiments was tested in vitro by apple polyphenols,blackcurrant anthocyanin,lycium ruthenicums anthocyanins,blueberry anthocyanins and apple polysaccharides on Classical Swine Fever Virus(CSFV)in this study.Methods:(1)Morphological observation and MTT colorimetric assay were used to determined the effects of different concentrations of polyphenol polysaccharides on the proliferation of PK-15 cells and their maximum safety concentration.(2)To study the antiviral effects of plant polyphenols and polysaccharides,the cellular active and proliferation of CSFV on PK-15 cells was identified by MTT colorimetry and RT-PCR in this experiment.(3)The following treatments were performed on the cultured PK-15 cells to research the antiviral action of polyphenol polysaccharides in different treatment ways: First,the cells were dropwise added the polyphenols polysaccharides then inoculated virus to inhibit adsorption of virus.Second,the cells were inoculated virus then dropwise added the polyphenols polysaccharides to inhibit replication of virus.Third,the cells were inoculated virus that had been dropwise added the polyphenols polysaccharides to direct inactivated virus.(4)The mouse macrophages were used to understand the effect of different polyphenol polysaccharides on cytokines,such as IFN-γ,TNF-α and IL-1α cytokines were detected by ELISA Kit.Results:(1)Maximum safe concentrations of each polyphenol polysaccharide was detected by morphological observation and MTT colorimetric assay: apple polyphenols 20μg / m L,apple polysaccharide 20μg / m L,black currant anthocyanin 40μg / m L,blueberry anthocyanin 50μg / m L,lycium ruthenicum anthocyanin 100μg / m L,the results of MTT and OD T-test manifest that is no difference with the control group obviously(p> 0.05).(2)The proliferation of CSFV on PK-15 cells was determined by MTT colorimetric method and RT-PCR,which provided experimental conditions for the antiviral activity of plant polyphenols and polysaccharides.(3)According to the results determined by MTT method and the OD value T test analysis showed that apple polyphenol,black wolfberry anthocyanin,blueberry anthocyanin,apple polysaccharide to directly killing effect of CSFV,therapeutic index respectively16,16,16,80,16,was significantly difference with control(p <0.05);blueberry anthocyanin,black currant anthocyanin,anthocyanins black wolfberry and other CSFV having adsorption inhibition effect,the therapeutic index was 80,16,16,the control group were significantly different(p<0.05);wolfberry anthocyanin and black apple polyphenol can inhibit the replication of CSFV,therapeutic indices are 16,16,and the control group were significantly different(p<0.05).(4)The result of Cytokine ELISA kit showed Apple polysaccharide(20μg / m L)in mouse macrophages can promote TNF-α,IFN-γ was produced,black currant anthocyanin(12.5μg / mL)promotes small produce IL-1α murine macrophage cells,and the control group were significantly different(p <0.05).Conclusion:(1)The results of antiviral tests in vitro showed that blueberry anthocyanin,black anthocyanin,and black galenic anthocyanins have the effect of inhibiting virus adsorption;apple polyphenols,black gallon anthocyanins,black anthocyanins,Blueberry anthocyanin,apple polysaccharide have the effect of directly killing viruses;black anthocyanins,apple polyphenol have the effect of inhibiting virus replication.(2)Apple polysaccharide can promote the secretion of TNF-α and IFN-γ from mouse macrophages.Black gallon anthocyanins can promote the secretion of IL-1α from mouse macrophages.