摘要
为了研究空心莲子草醇提物[Alternanthera philoxeroides(Mart.)Griseb.alcohol extract,AAE]的体外抗H9N2型禽流感病毒(avian Influenza virus,AIV)的效果,探索空心莲子草体外抗病毒的作用方式,试验测定了AIV-H9N2对DF-1细胞的TCID50;采用观察细胞病变联合CCK-8检测的方法,测定AAE对DF-1细胞的毒性作用;通过计算各试验组(空白对照组、病毒对照组、给药组)细胞的存活率,观察3种作用方式(预防感染作用、抑制复制作用及直接杀灭病毒作用)下,不同浓度AAE体外抗AIV-H9N2的效果,并计算治疗指数(TI)。经尿囊腔接种病毒,测定鸡胚的半数胚致死量(ELD50),测定AAE对鸡胚的最大安全浓度,然后对鸡胚进行攻毒用药试验,统计各组存活情况及血凝效价,观察胚体的发育情况。结果表明:收获的AIV-H9N2病毒液TCID50为1×10^(-4.23)/0.1 mL;AAE对DF-1细胞作用24,48小时时的CC50浓度分别为(191.52±8.73)μg/mL、(156.96±8.79)μg/mL;3种作用方式下,药物对细胞的保护率范围分别在40%~58%、4%~34%及12%~18%之间,AAE治疗指数预防病毒感染作用(3.42±0.71)大于影响病毒复制作用(1.74±0.11),AAE不具有直接杀灭病毒作用。鸡胚的ELD50为1×10^(-2.84)/0.2 mL,AAE对鸡胚的最大安全浓度为32.5 mg/mL,AAE能显著改善AIV-H9N2感染胚胎的存活情况(P<0.05),降低病毒感染的血凝效价,减弱病毒对胚体发育的影响。说明AAE在DF-1细胞和鸡胚中对AIV-H9N2具有潜在的抑制作用,可以抑制AIV-H9N2的活性。
In order to research the in vitro effect against avian Influenza A virus H9N2(AIV) of Alternanthera philoxeroides(Mart.) Griseb. alcohol extracts(AAE), and explore the in vitro antiviral mode of action of Alternanthera philoxeroides(Mart.) Griseb., TCID50of AIV-H9N2 on DF-1 cells was determined in the experiment. The toxic effect of AAE on DF-1 cells was determined by using observation of cytopathic lesions combined with CCK-8 assay. By calculating the cell survival rates of each test group(cell blank control group, virus control group, drug administration group), the effects of different concentrations of AAE against AIV-H9N2 in vitro under three modes of action(preventive effect on virus infection, affecting virus replication, direct inactivation of virus) were observed, and the therapeutic index(TI) was calculated. The virus was inoculated via the urinary bladder cavity, and the ELD50value of chicken embryos was determined;the maximum safe concentration of AAE on chicken embryos was determined;then the chicken embryos were subjected to a challenge-medication test, and the survival and hemagglutination titer of each group were counted;the development of embryos was observed. The results showed that the TCID50of the harvested AIV-H9N2-virus solution was 1×10^(-4.23)/0.1 mL;the CC50concentrations of AAE on DF-1 cells at 24 and 48 h were(191.52±8.73),(156.96±8.79) μg/mL, respectively. Under the three modes of action, the cell protection rates of the drugs ranged from 40% to 58%, 4% to 34% and 12% to 18%, respectively. The AAE therapeutic index preventing viral infection(3.42±0.71) was more than affecting viral replication(1.74±0.11), and AAE did not have a direct virucidal effect. The ELD50value of chicken embryo was 1×10^(-2.84)/0.2 mL, and the maximum safe concentration of AAE for chicken embryo was 32.5 mg/mL. AAE significantly improved the survival of AIV-H9N2-infected embryos(P<0.05), reduced the hemagglutination titer of viral infection, and attenuated the effect of the virus on embryo development. The results suggested that AAE had a potential inhibitory effect on AIV-H9N2 in DF-1 cells and chicken embryos, which could inhibit the activity of AIV-H9N2.
作者
郑新
戴琪
李永华
方佳慧
刘焱文
张秀桥
ZHENG Xin;DAI Qi;LI Yonghua;FANG Jiahui;LIU Yanwen;ZHANG Xiuqiao(College of Pharmacy,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Provincial Key Laboratory of Resources and Chemistry of Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430065,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2023年第5期117-122,138,共7页
Heilongjiang Animal Science And veterinary Medicine
基金
湖北中医药大学2021年“青苗计划”项目(2021ZZX020)。
作者简介
郑新(1999-),女,硕士研究生,研究方向为中药资源品质评价与开发,1941249583@qq.com;通信作者:张秀桥(1965-),女,教授,博士,研究方向为中药资源品质评价与开发,qiaoxzh@163.com.
