小檗胺影响牛病毒性腹泻病毒体外复制的研究

This study was conducted to investigate whether small molecule drug Berbamine (BBM) affected Bovine viral diarrhea virus (BVDV) replication in vitro. Preparations of BBM at 0, 1, 5, 8, 10, 15 and 20 μmol/L concentrations were added to MDBK cell cultures and the cells were infected with BVDV 8 h later. MTT assay was performed to detect the effect of BBM on the proliferation of MDBK cells and optimal concentration. The accumulation of BVDV double strand RNA (dsRNA) was tested in BBM treated MDBK cells in immunofluorescence staining. At the same time, quantitative real-time PCR and Reed-Muench method inverted microscope were applied to determine the effect of BBM on BVDV RNA replication, viral titer alteration and CPE. The results showed as low as 10 μmol/L of BBM significantly inhibited the accumulation of BVDV dsRNA, and the cell toxicity was small at this concentration. Quantitative real-time PCR and titer assay showed that the level of 5'UTR mRNA and viral titer in MDBK cells treated with BBM were significantly lower than control at 72 h post inflection and the most difference of virus titers between BBM treated or non-treated cells was presented at 24 h and 36 h post infection. In addition, BVDV infection caused severe CPE in non-treated MBDK cells but less CPE in BBM-treated cells. In conclusion, BBM treatment significantly inhibited BVDV replication in cells.