| The swine pseudorabies(PR)caused by Pseudorabies virus(PRV)is characterized by reproductive disturbance,dyspnea,neurological symptoms,and so on.It is susceptible to pig herds at all ages and has caused huge losses to the pig industry.Because of the emergence of PRV mutant strains in recent years,the pigs can not be completely protected from PRV infection by immuning with conventional vaccines.So it is important to study the antiviral drugs.In this study,the antiviral effect and working time of hydroquinone on PRV in vitro and was studied on PK-15 and N2a cells.Finally,the effect of hydroquinone was confirmed in mice through inoculating with PRV ZJ01.This study preliminarily revealed the antiviral effect of hydroquinone on PRV,and provided some reference for the prevention and treatment of PRV.Specific research contents are as follows:1.Screening of anti pseudorabies virus drug and its effect in vitroIn order to screen the anti-PRV drugs,44 synthetic chemicals in FDA-approved drug library were used on PK-15 cells and screened by IFA.The results showed that hydroquinone had antiviral effect on PRV.IC50 of hydroquinone and SI of PRV on PK-15 and N2a cells were measured.The results showed that the maximum no-cytotoxic concentration of drug,IC50 and SI were 30μM,5.113μM,18.78 in PK-15 cells and 18μM,8.338μM,11.14 in N2a cells.To confirm the function of hydroquinone,the working dose,time and the anti-different PRV strain function of hydroquinone were detected on PK-15 and N2a cells by IFA,Western-blot and qPCR.The results showed that there was a dose-dependent increase in anti-PRV effect of hydroquinone and the working time of hydroquinone was different on PK-15 and N2a cells.The hydroquinone could inhibit virus adsorption on PK-15 cell and virus adsorption and cell entry on N2a cells.This study firstly revealed that the hydroquinone has antiviral effect on PRV in vitro.2.Effect of hydroquinone on pseudorabies virus in vivoIn order to study the anti-PRV effect of hydroquinone in vivo,ICR mice were used in the study.Fifty-eight mice were randomly divided into three groups:drug-treated group,non-drug-treated group and blank control group.The mice of the administration group and the non-administration group were injected with 5LDso PRV ZJ01 on the back,and the mice of blank control group was injected with the same amount of DMEM.The mice in the administration group were injected with 50 mg/kg hydroquinone intraperitoneally 8 hours after challenge,and then given same dose of drug every 12 hours.The mice in the other two groups were injected with the same amount of saline at the same time.The symptoms of each group of mice were observed,and the time of death and mortality was recorded.Seven mice in each group were killed 48 hours and 60 hours after challenge.The pathological changes were observed and the viral load in tissues and organs was measured by qPCR.The results showed that the mice in the non-administration group began to die on the 3rd day,the mortality rate was 37.5%,the 4th day was 80%,and all the mice died on the 5th day.The death time of mice in the administration group was delayed to the 4th and 5th day.On the 4th day,the mortality rate of mice in this group was 50%,and on the 5th day,the mortality rate was 50%,and two mice survived in the end of experiment.Pathological changes showed that severe hemorrhage,congestion and inflammatory infiltration were found in the brain tissue of mice in the non-administration group under the pathological microscope.While the brain tissue of mice in the administration group was normal but only slight hemorrhage.At 48h and 60h after challenge,the viral load in the brain of mice in the drug-treated group was significantly lower than that in the non-drug-treated group,and the difference was most obvious at 60h.These results indicated that hydroquinone has anti-PRV function in ICR mice. |
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