鬼针草水提物抗伪狂犬病毒作用研究

Effect of aqueous extract of Bidens pilosa L.on Pseudorabies virus

【目的】研究鬼针草水提物(BPE)的体外抗伪狂犬病毒(Pseudorabies virus,PrV)作用。【方法】制备640 mg/mL(以生药计)的BPE,检测其对仓鼠肾上皮细胞(BHK-21)的毒性(半数中毒浓度(TC_(50)))及对PrV体外抗性(半数抑制浓度(EC_(50)))和治疗指数(TI)。采用CCK-8法检测BPE抗PrV的机制。采用间接免疫荧光法检测BPE对gB蛋白表达和PrV所致细胞凋亡的影响,采用实时荧光定量PCR方法检测BPE对PrV gB基因表达的影响。【结果】BPE对BHK-21细胞的TC_(50)为28.7 mg/mL,对PrV的EC_(50)为5.16 mg/mL,TI为5.56。BPE抗PrV的机制为抑制PrV增殖和灭活PrV。BPE可抑制PrV诱导的BHK-21细胞凋亡,抑制PrV gB基因和蛋白的表达。【结论】BPE具有明显的体外抗PrV活性。

【Objective】This study aimed to investigate the protective effect of aqueous extract of Bidens pilosa L.(BPE)on Pseudorabies virus(PrV)in vitro.【Method】The cytotoxicity and anti-PrV activity of BPE at concentration of 640 mg/mL on BHK-21 cells were detected by CCK-8,and the anti-PrV mechanism of BPE was investigated by CCK-8.The TC_(50),EC_(50) and TI were determined.The indirect immunofluorescence was used to detect the effect of BPE on PrV-induced apoptosis and expression of PrV gB protein,and the real-time fluorescence quantitative PCR was used to detect the expression of PrV gB gene.【Result】The TC_(50) of BPE in BHK-21 cells was 28.7 mg/mL,the IC_(50) of PrV was 5.16 mg/mL,and the TI was 5.56.The mechanism of anti-PrV was based on inhibiting PrV proliferation and inactivating PrV.BPE might protect PrV-infected BHK-21 cells through suppressing expression of PrV gB gene and protein(P<0.01)and inhibiting PrV-induced apoptosis of BHK-21 cells.【Conclusion】BPE had significant anti-PrV activity in vitro.