| Bovine viral diarrhea virus(BVDV),bovine enterovirus(BEV)and equine herpesvirus-1(EHV-1)induce different degrees of symptoms in sick animals,affecting the healthy development of breeding industry,and there are no specific therapeutic drugs for virus infection.Berbamine(BBM)have the effects of antiinflame and anti-cancer,regulating blood pressure,revitalizing necrotic tissue and other effects.As a calmodulin antagonist,BBM can significantly inhibit the proliferation of tumor cells and regulate myocardial infarction induced by isoproterenol by improving mitochondrial function.However,the application of BBM in antiviral research has not been reported.In this study,BBM was applied in antiviral therapy,to explore the effects of BBM on BVDV,BEV,EHV-1 replication,which provides new technology and means for BVDV,Bev,EHV-1 prevention and treatment.Method: MTT method was applied to detect the effect of different concentration of BBM on the proliferation of MDBK cells,and determined the optimal concentration of BBM.MDBK cells were treated with different concentrations of BBM,and following infected with BVDV and BEV,the accumulation of double strand RNA(ds RNA)was detected by immunofluorescence staining.The effects of BBM on BVDV and BEV RNA,EHV-1 DNA replication,titer alter,cytopathic effect(CPE)were detected by quantitative real-time PCR(q RT-PCR),Reed-Muench method and inverted microscopy.The effect of BBM on BVDV and BEV infection in BALB/c mice was divided into the preventive protection test and the therapeutic test.BALB/c mice(3 ～4 months)were randomly divided into two groups,and the control group was given 0mg/kg BBM every day,while the experimental group was given 50 and 100 mg/kg BBM every day by gavage.After gavage for 3 d,BVDV or BEV was used to infect BBM treated BALB/c mice for three times.After virus infection,BBM was administered by gavage every 24 h.At 5 and 10 d after infection,the pathological tissues were dissected and made into sections,and following with H&E staining.q RT-PCR was performed to detect the replication of viral RNA in mice.In the experimental group,BALB/c mice(3 ～ 4 months)were randomly divided into two groups,After BVDV or BEV infection for 3 d,the control group was given 0mg/kg BBM per day,while the experimental group was given 100 mg/kg BBM per day.At 0,3,6 and 10 d after virus infection,the tissues were dissected and made into sections,and histopathologic changes were observed after H&E staining.The replication of viral RNA in mice was detected by q RT-PCR.Result: In vitro the m RNA or DNA levels of BVDV,BEV or EHV-1 in host cells treated with BBM were50-fold lower than those in the control group,and progeny virus titer were significantly decreased.The change of cell morphology showed that BBM could attenuate CPE induced by BVDV,BEV or EHV-1infection,indicating that BBM treatment was significantly prevented BVDV,BEV,EHV-1 replication in vitro.The result of in vivo experiment showed that BBM treatment significantly inhibited BVDV,BEV replication in the preventive protection test and the therapeutic test,and BBM treatment significantly inhibited the histopathologic changes and reduced the viral load of each tissue.Conclusion: BBM significantly reduced BVDV,BEV and EHV-1 replication,which is expected to be a potential drug for the prevention and treatment of calf diarrhea and equine herpes viral abortion. |
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