摘要
阿瑞吡坦具有抗肿瘤和抗HIV-1活性。为了探究与非洲猪瘟病毒(ASFV)D1133L亲和力较高的阿瑞吡坦是否具有抗ASFV作用,利用MTT方法测定阿瑞吡坦对PAMs细胞的毒性,然后将荧光蛋白重组ASFV感染PAMs细胞后观察荧光变化和TCID_(50)、同时用RT-qPCR和Western-blot分析阿瑞吡坦对ASFV复制及D1133L、P30和P72转录和翻译的影响。结果表明,当DMSO处理PAMs时,细胞存活率未达到100%,对PAMs细胞具有一定毒性;当浓度为20和40μmol/L的阿瑞吡坦处理PAMs后,PAMs细胞存活率均接近100%,当浓度为80μmol/L的阿瑞吡坦处理PAMs时,细胞存活率达到70%以上:当阿瑞吡坦处理感染荧光蛋白重组ASFV的PAMs细胞后,ASFV的荧光值、基因组拷贝数和TCID_(50)被显著降低,ASFV D1133L、P30和P72转录和翻译被显著抑制。结论:阿瑞吡坦在体外对PAMs细胞的毒性较小,通过靶向ASFV编码的D1133L蛋白显著抑制ASFV的体外复制,对ASFVP30和P72的转录和翻译也具有抑制作用。
Aprepitant has anti-tumor and anti-HIV-1 activities.In order to explore whether aprepitant,which has high affinity with African swine fever virus(ASFV)D1133L,has anti-ASFV effect,the cytotoxicity of aprepitant was determined by MTT method,and then the fluorescence changes and TCID_(50)were observed after cells were infected with fluorescent protein recombinant ASFV,and RT-q PCR and Western-blot experiments were used to analyze the effect of aprepitant on ASFV replication,D1133L,P30 and P72 transcription and translation.The results showed that the cell survival rate did not reach 100%when DMSO treated PAMs,mean DMSO had a certain degree of toxic to PAM cells.The cell survival rate was close to 100%under 20 and 40μmol/L aprepitant condition,and the cell survival rate was more than70%under 80μmol/L aprepitant condition.When the PAM cells infected with fluorescent protein recombinant ASFV were treated with aprepitant,the fluorescence value,genome copy number and TCID_(50)of ASFV were significantly reduced,and the transcription and translation of ASFV D1133L,P30 and P72 were significantly inhibited.In conclusion,aprepitant was less toxic to PAM cells in vitro,and can significantly inhibited ASFV replication in vitro by targeting the D1133L protein encoded by ASFV,and also inhibited the transcription and translation of ASFV P30 and P72.
作者
崔卉梅
史喜绢
张大俊
杨行
杨博
杨金柯
郝雨
陈玲玲
闫文倩
刘湘涛
郑海学
张克山
CUI Hui-mei;SHI Xi-juan;ZHANG Da-jun;YANG Xing;YANG Bo;YANG Jin-ke;HAO Yu;CHEN Ling-ling;YAN Wen-qian;LIU Xiang-tao;ZHENG Hai-xue;ZHANG Ke-shan(State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Animal Virology,Ministry of Agriculture and Rural Affairs/National Foot-and-Mouth Disease Reference Laboratory/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou,730046,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2022年第10期1207-1214,共8页
Chinese Veterinary Science
基金
国家重点研发计划项目(2021YFD1801300)
甘肃省科技重大专项(20ZD7NA006)
中国农业科学院基本科研业务费项目(Y2020XK09)
