Effect Of Probiotic Increased Galactose On The Inhibition Of RV Infection

The glycoprotein or glycolipids on the surface of eukaryotic cells contain galactosylated glycans.Studies have shown that galactose is not only extensively involved in the synthesis of various sugar chains,but also acts as terminal residues and galactose residues,Adhesion,migration and growth.Rotaviruses(RVs)need to recognize cell surface glycans in the process of attachment to intestinal epithelial cells,so galactose is closely related to the process of rotavirus-infected cells.To clarify whether the supernatant of probiotics can increase the effect of anti-RV infection by inducing the increase of galactose on the surface of intestinal mucosa cells.In this study,three different probiotic supernatants were used to incubate the small intestinal epithelial cells(IPEC-J2)to induce three differentiated IPEC-J2 s.The role of probiotics in anti-RV infection was tested with differentiated IPEC-J2.At the same time,the use of D-galactose instead of high glucose DMEM culture medium in the culture of IPEC-J2 cells to obtain a new differentiation type,and probiotics induced differentiation type of cell surface galactosylation degree and anti-RV ability.In this study,the cells were divided into five groups: Lactobacillus casei(group g),Bacillus subtilis induced group(k),Lactobacillus amylovorus induction group(s group),galactose induced group(gal group),and Untreated cells.The above treatment group was divided into porcine rotavirus(PRV)infected cells and uninfected cells.The amount of galactose on the surface of IPEC-J2 cells was observed by agglutinin fluorescence technique.Using the Reed-Muench method detected each infected cell group's half of the amount of tissue infection of rotavirus.The amount of rotavirus and ?-galactosidase and?-galactosyltransferase in each infected group were measured by radioimmunoassay(ELISA)at12h,24 h,48h and 72 h after infection.The untransfected cells of each group were used as control.From the lectin fluorescence diagram,it can be seen that the supernatant of three probiotics and the incubation of galactose alone could alter the content of galactose on the cell surface.Half of the infection test results show that when the rotavirus infection 72 h,the untreated cells p RV TCID50 / 0.1 m L of 10-7.15 ± 0.14,g group p RV TCID50 / 0.1m L was 10-3.875 ± 0.125,s group pRV TCID50 / 0.1 mL was 10-4.125 ± 0.138,k group pRV TCID50 / 0.1m L was 10-4.16 ±0.144;virus quantitative test results showed that RV from untreated cells proliferated slowly in12 h to 24 h after infection,RV proliferated rapidly from 24 h to 48 h and the virus quantity reached the peak,from 48h-72 h virus number decreased.The RV group was significantly lower than that of any group of cells 24 hours after RVinfection.It is suggested that the antiviral effect of galactose is most obvious during viral replication.The RV content of the three groups of probiotics was significantly lower than that of the untreated group within 48 hours of RV infection.There was no significant difference in RV content between the three groups(P?0.05).The concentration of ?-galactosidase in normal cells was higher than that in group g,k and s(P <0.05)in uninfected cells.S group and k group were significantly lower than normal cells and g group at 12 th hour.group s and k were significantly lower than those in normal and g groups at24 th hour..The concentration of enzyme in group s and g was significantly lower than that in normal and k at 48 th hour.The s and k groups were significantly lower than those in normal and g groups at 72 h after RV infection(P <0.05)There was no difference between groups k and s(P ?0.05).Before RV infection,the concentration of galactose transferase in normal cell was significantly lower than those in group g,group k and group s(P<0.05).In 12 hours after RV infection,the concentration of galactose transferase of group k,group g and group s were significantly higher than that of normal cells(P<0.05).In 24 hours after RV infection,The concentration of group g,group k and group s group were significantly higher than that in normal group(P<0.05).In 48 hours after RV infection,the concentrations of group g and group s were significantly higher than that of normal cells(P<0.05).As mentioned above,Lactobacillus casei supernatant,Lactobacillus amylovorus supernatant,Bacillus subtilis supernatant can increase the galactose content of IPEC-J2 cell surface to prevent the adhesion of RV to cells and enhance the ability of anti-RV cells,thus anti-RV infection that can protect the cells.