绞股蓝皂苷及其磷酸化修饰物体外抗DHAV作用的比较  

Comparison of gypenosides and its phosphorylated derivative against DHAV in vitro

作　　者：白景英 韩开顺 杜红旭[1] 熊文 张伟[1] 明珂[1] 王金丽 袁文娟 刘家国[1] BAI Jing-ying1 , HAN Kai-shun2 ,DU Hong-xu1 ,XIONG Wen1 ,ZHANG Wei1 ,MING Ke1 ,WANG Jin- li1 , YUAN Wen-juan1 , LIU Jia-guo1(1. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China; 2. Pingyuan Animal Husbandry and Veterinary Bureau, Dezhou, Shandong 253100, Chin)

机构地区：[1]南京农业大学动物医学院,江苏南京210095 [2]山东省平原县畜牧兽医局,山东德州253100

出　　处：《中国兽医学报》2018年第5期978-985,共8页Chinese Journal of Veterinary Science

基　　金：中央高校基本科研业务费资助项目(Y0201700441);国家自然科学基金资助项目(31572557,31772784);公益性行业(农业)科研专项(201303040-4,201403051);江苏高校优势学科建设工程项目(PAPD)

摘　　要：采用三聚磷酸钠和三偏磷酸钠法制备磷酸化绞股蓝皂苷（pGP），然后运用体外细胞培养法比较研究绞股蓝皂苷（GP）和pGP抗DHAV感染鸭胚肝细胞（DEH）的作用。通过MTT法测定了GP和pGP在DEH上的安全质量浓度，并采用先加药物后接种病毒、先接种病毒后加药物和药物与病毒同时感作3种加药方式观察了GP和pGP对DHAV（duck hepatitis A virus）感染DEH的影响。为了进一步验证药物的抗病毒效果，采用实时荧光定量PCR方法比较分析了最有效药物浓度的GP和pGP对DHAV在DEH上的吸附、复制和释放的影响。结果显示：GP和pGP在一定质量浓度下均能促进DEH的生长；GP和pOP都具有较好的体外抗DHAV的作用，且在先加药后加毒和先加毒后加药作用方式下，pGP的抗DHAV的作用效果优于GP；在先加药后加病毒作用方式时，GP和pGP均能有效抑制DHAV的吸附；在先加病毒后加药作用方式时，GP和pGP对病毒吸附鸭胚肝细胞没有明显的作用，但能有效抑制DHAV的复制和释放，且pGP的抑制效果优于GP。结果表明：磷酸化修饰显著提高了GP的抗DHAV作用，pGP有希望被开发成一种新型抗DHAV药物。In order to research and develop a new effective drug against duck hepatitis A virus （DHAV）. Phosphorylated derivative of gypenosides （pGP） was prepared by the sodium trime- taphpsphatesodium tripolyphosphate method, then antiviral activity against DHAV was studied by duck embryonic hepatocytes （DEH） culture in vitro. MTT colorimetric assay was used to detect the effects of gypenosides （GP） and pGP on the growth of DEH. The saponins and DHAV were added into DEH by the following three ways.adding saponins firstly,adding virus firstly and adding saponins and virus simultaneously to observe their anti-virus functions. To further verify the antiviral effect of the drugs,the real-time polymerase chain reaction（ real-time PCR） was ap plied to study the antiviral mechanism of adsorption, replication and release. The results showed that GP and pGP within safe concentrations could promote the proliferation of DEH. Both GP and pGP within safe concentrations had good anti-DHAV effect in vitro ,and pGP performed more ef fectively than GP when adding saponins firstly and adding virus firstly, respectively; Both GP and pGP could effectively inhibit the adsorption of DHAV when adding saponins firstly; when adding virus firstly,GP and pGP did not show any influence on the adsorption of DHAV,but they could effectively inhibit the replication and release of DHAV, and pGP was more effective than GP. Phosphorylated modification significantly improves the anti-DHAV effect of GP,and pGP is promising to develop into a novel anti-DHAV drug.

关 键 词：DHAV 绞股蓝皂苷 磷酸化修饰 抗病毒活性 

分 类 号：S852[农业科学—基础兽医学]