Studies On The Effect Of Curcumin Prevention And Treatment On RABV Virus

Objective:With the attempt to study the preventing and treating effect of Curcumin on BD06 isolate, we used vivo and vitro experiments to explore its immune mechanisms. Provide theoretical basis for clinical application. Method:Vitro cell experiment and vivo animal experiment were used in this study. In vitro trials, MTT was used to investigate the effect of curcumin on the profilation of BHK-21; the preventing and treating effect were determined by direct immunofluorescence assay(DFA) after interfered with different concentrations of Curcumin. In vito trials, 14-16 g Kunming mice were randomly divided into eight groups: control, model, DMSO, Vaccine, Cur 25mg/kg, Cur 50mg/kg, Cur 100mg/kg, Vaccine+Cur 50mg/kg. Influence of curcumin on the incubation period of mice infected by BD06 isolate was infirmed by DFA; the gene level of IFN-β, GAPDH in the peripheral blood were detected by Real time-PCR in the 2 days; the growth characteristics of BD06 were detected by spinal cord and brain tissue RT-PCR in the 3 days, 5 days. Result:(1) Effects of curcumin on the profiration of BHK-21. The results showed that with the increase of concentration, 60mM, 80mM can significantly inhibit cell viability(P<0.05) in contrast with the control. However, the cell viability in the other groups were not significantly changed(P>0.05).(2) Treating effect of Curcumin on BD06 isolate in vitro. Compared with the control, fluorescence intensity changed slightly, which means DMSO may have no antiviral effect. In the group of Cur 5mM, 10mM, 20mM, 40mM, the fluorescence intensity receded gradually, especially in the Cur 40mM. The differences between control group and Cur 2.5mM、Cur 5mM were no significant.(3) Preventing effect of Curcumin on BD06 isolate in vitro. The DFA results showed that a number of infected cells can be detected in the control group and DMSO group, indicating that pretreatment with DMSO had no effect on virus infection. The fluorescence intensity were strong and clear cell morphology could be seen in the Cur 2.5mM. However, in the group of Cur 5mM, 10mM, 20mM, 40mM, the intensity weaken gradually, indicating that the number of infected cells gradually decreased.(4) Effect of curcumin on the incubation period of mice infected by BD06 isolate. The results showed that compared with the model group, Cur50, Cur100, Vac+Cur50 can obviously delay the incubation period(P<0.01), while the Cur25 and Vac can not prolong incubation period significantly(P>0.05). Compared with the Cur50, The incubation period between Vac+Cur50 and Cur100 had no significant different(P>0.05).(5) Influence of curcumin on the growth characteristics of BD06. The results of RT-PCR showed that the spinal cord can detected RABV in the model, Vac, Cur25, but RABV in the Cur50, Vac+Cur50 cannot be detected by RT-PCR in 3 days after intramuscular. The corresponding strap cannot be detected in the cerebellum after RT-PCR in each group.Agarose gel electrophoresis showed that in 5 days after intramuscular, except the Cur50, spinal cord in the each group can detected the DNA strap. Compared with the results in 3 days, DNA srap can be detected distinctly in 5 days, especially on the sample 12 th of control(6) Effect of curcumin on the level of IFN-β in the mice infected with BD06. Real time-PCR showed that in the second day after infection, the DMSO group cannot effect the gene level of IFN-β. However, in contrast with the Vac, the gene level of IFN-β in Vac+Cur50 and Cur50 significantly improved(P<0.05); however, compared with the Cur25, the gene level of IFN-β significantly improved in Cur50(P<0.05). Conclusion:(1).A certain dose of curcumin have prevention and treatmen effect on the RABV in vitro experiment, especially in the dose of 40mmol/L;(2). Curcumin can prolong the incubation period of mice infected by RABV to 10 days, delaying the onset time of 2-3 days;(3). Curcumin can improve the gene level of IFN-β in the mice infected by RABV.