金丝桃素体外抗传染性支气管炎病毒的研究  

Study on antiviral effect of hypericin against infectious bronchitis virus in vitro

作　　者：陈惠杰 刁磊[2] 张岳[1] 刘海鑫 李伟群 黄小丹 张瑞莉[1] 李广兴[1] CHEN Hui-jie;DIAO Lei;ZHANG Yue;LIU Hai-xin;LI Wei-qun;HUANG Xiao-dan;ZHANG Rui-li;LI Guang-xing(College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China;College of Pharmaceutical Engineering,Jilin Agriculture Science and Technology University,Jilin City 132101,China)

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]吉林农业科技学院制药工程学院,吉林吉林132101

出　　处：《中国兽医科学》2018年第11期1415-1422,共8页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31172295,31272569)

摘　　要：将不同浓度的金丝桃素和鸡传染性支气管炎病毒（IBV）以三种方式加入鸡胚肾细胞（CEK）培养体系中,采用细胞病变效应（CPE）法、MTT法、病毒滴度法、实时荧光定量PCR以及间接免疫荧光技术检测金丝桃素体外抗IBV作用,同时检测其对IBV诱发细胞凋亡的影响。结果显示,金丝桃素在CEK细胞感染IBV前、后及同时加入,都有明显抑制CPE的作用;随着添加药物浓度的增加,CEK细胞活力明显增高,病毒TCID50滴度、病毒核衣壳蛋白RNA相对表达量和CEK细胞中病毒阳性绿色荧光信号显著降低或减少（P〈0.01）,并且呈现明显的剂量依赖性。三种给药方式以药物和病毒互作后同时加入效果最好。IBV可以引起CEK细胞凋亡的发生,添加金丝桃素能够抑制病毒引起的凋亡。上述试验结果表明,金丝桃素具有很好的抗IBV作用,其作用可能是通过抑制感染细胞凋亡产生的,这可能是金丝桃素抗IBV的作用机制之一。Hypericin of different concentrations and IBV were added to the culture system of chicken embryo kidney（CEK） cells in three ways,cytopathic effect（CPE） observation,virus titer assay,MTT method,real-time fluorescence quantitative PCR and indirect immunofluorescence were used to detect the anti-IBV effect of hypericin in vitro.At the same time,the apoptotic morphological changes of CEK cells post-addition of hypericin to IBV were detected with apoptosis kit.In result,hypericin could significantly inhibit the CPE in all three ways of adding hypericin before,after or simultaneously with IBV into CEK cells.With the increase of hypericin concentration,the viability of CEK cells increased remarkably,the virus titer of TCID50,the relative expression of IBV N gene,and the positive fluorescence signal of the virus in CEK cells decreased significantly（P〈0.01）.The anti-viral effect of hypericin was obviously concentration dependent.Among three ways of addition of the drug,the most effective way was addition of hypericin into IBV simultaneously post-incubation.Hypericin could inhibit the apoptosis caused by IBV.In conclusion,hypericin had good anti-IBV effect and its function may be mediated by inhibiting the apoptosis of infected cells,which may be one of its anti-IBV mechanisms.

关 键 词：金丝桃素 鸡传染性支气管炎病毒 抗病毒活性 CEK细胞 抗IBV 

分 类 号：S852.659.6[农业科学—基础兽医学]