Researches Of Yupingfeng Polysaccharides Antiviral Mechanisms In Vitro

Chinese medicine in livestock and poultry disease prevention and treatment plays an important role,especially for some immunosuppressive viral diseases and stress diseases have unique advantages.As the antiviral chemical treatment drugs have been banned in food animals,and thus carry out anti-viral Chinese medicine research highlights its importance and urgency.YuPingFeng polysaccharides(YuPingFeng polysaccharides,YPF-P)is an effective part of the total polysaccharide of Yupingfeng powder,which has the function of enhancing the immunity and inhibiting the virus infection.At present,a large number of studies have studied the mechanism of polysaccharide from Yupingfeng from humoral immunity and cellular immunity,but the anti-virus mechanism of Yupingfeng polysaccharide is still rarely reported,and it is urgent to strengthen its research.In this project,the concept of serum pharmacology was introduced.The antiviral study of Yupingfeng polysaccharide and its serum was carried out at home and abroad for the first time.The chick embryo and chicken embryo fibroblasts were infected with Newcastle disease virus(NDV)-P polysaccharide and its serum-containing antiviral mechanism.1.YPF-P was extracted by water extraction and alcohol precipitation,purified by macroporous resin and freeze-dried by vacuum.2.Preparation of serum containing serum and cytotoxicity:The optimal dose of YPF-P was administered to SPF rabbits,and whole blood was collected at 1,3,5 and 7days to prepare different concentrations of serum.Primary SPF chicken fibroblasts were cultured and different Concentration of YPF-P and YPF-P containing serum,the cytotoxicity test,observed cell lesions,detection of cell activity.3.Anti-NDV effect of serum containing HCVF-P and YPF-P:The anti-NDV effect was detected by three different modes of administration.The blocking,inhibition and inhibition of YPF-P and YPF-P sera were observed respectively.Direct killing effect;culture of primary SPF chicken fibroblasts,inoculated NDV virus before and after the same time,respectively,cytotoxicity experiments to observe the cell lesions,MTT assay cell activity.4.To investigate the effect of cryopreservation on the apoptosis and apoptosis of CEF cells:To investigate the effect of cryopreserved cells on the expression of Caspase-3,P53,Bcl-2/Bax,TNF?and other apoptotic regulatory genes.The purification rate of YPF-P polysaccharide was 88.9%.NDV chicken embryo half of the lethal concentration of 1:10~7.Compared with the normal control group,chick embryo fibroblast activity of different concentrations of YPF-P(0.625,1.25,2.5,5,10 mg/mL)and YPF-P-containing serum(0,1,3,5,7 d)(P<0.05 or 0.01),and the cells grew well.(P<0.01).The cell viability of YPF-P-containing serum was lower than that of normal and positive control group(P<0.01),and the activity of YPF-P was significantly higher than that of NDV control group(P<0.01),the cell viability of the drug-containing serum group was significantly higher than that of the NDV control group(P<0.01),and the difference was significant.YPF-P can block the occurrence of apoptosis by inhibiting the expression of apoptotic proteins.YPF-P polysaccharide and YPF-P-containing serum had no toxic and side effects on cells.YPF-P could block the adsorption and entry of NDV,and had good protective effect on chicken embryo fibroblasts.YPF-P-containing serum not only blocked the adsorption and invasion of NDV on chicken embryo fibroblasts,but also inhibited the toxicity of virus and disrupts the synthesis and release of NDV.The anti-NDV effect in vitro was higher than that of YPF-P.