人副流感病毒2型感染恒河猴肾细胞后内参基因的筛选  

Screening of internal reference genes after rhesus monkey kidney cells are infected with human parainfluenza virus 2

作　　者：袁明铭 周静[1] 夏介英[1] 董涵 洪杨[1] 张磊[1] YUAN Mingming;ZHOU Jing;XIA Jieying;DONG Han;HONG Yang;ZHANG Lei(Sichuan Academy of Chinese Medicine Sciences,Chengdu,Sichuan 610041,China)

机构地区：[1]四川省中医药科学院,成都610041

出　　处：《重庆医学》2020年第17期2861-2866,共6页Chongqing medicine

基　　金：四川省公益性科研院所基本科研业务专项项目(2016ysky0086);四川省科技基础条件平台项目(2018TJPT0044)。

摘　　要：目的评价阳性药物利巴韦林治疗人副流感2型病毒(HPIV-2)感染恒河猴肾细胞(LLC-MK2)后甘油醛-3-磷酸脱氢酶(GAPDH)、次黄嘌呤磷酸核糖转移酶1(Hprt1)、β-肌动蛋白(β-actin)、肽基脯氨酰异构酶A(Ppia)、β2微球蛋白(B2M)5种内参基因的表达稳定性。方法HPIV-2感染LLC-MK248 h和72 h,采用real-time PCR法测定病毒组、利巴韦林治疗组和正常细胞对照组中上述5种内参基因的表达水平,利用geNorm程序对它们的稳定性进行评价。结果5种内参基因均能稳定表达,其中GAPDH和Hprt1在感染后48 h稳定度相同,且最稳定;Ppia和Hprt1在感染后72 h稳定度相同,且最稳定。结论Hprt1、GAPDH和Ppia是抗HPIV-2药物体外筛选和研究HPIV-2与LLC-MK2相互作用最理想的内参基因。Objective The purpose of this study is to analysis the expression stability of glyceraldehyde-3-phosphate dehydrogenase(GAPDH),Hypoxanthine phosphoribosyl transferase 1(Hprt1),β-actin,peptidylprolyl isomerase A(Ppia),β2 microglobulin(B2M)genes in Rhesus kidney cells(LLC-MK2)infected by Human parainfluenza virus 2(HPIV-2)and treated by ribavirin.Methods LLC-MK2 were infected with HPIV-2 for 48 and 72 hours,then the expression levels of the above five internal reference genes in LLC-MK2 cells of the viral group,the ribavirin treatment group and the normal cell control group were determined by real-time PCR method,geNorm program was used to evaluate their stability.Results Five kinds of internal reference genes could be stably expressed after infection.Among them,GAPDH and Hprt1 had the same stability at 48 h after infection and were the most stable;Ppia and Hprt1 had the same stability at 72 h after infection and were the most stable.Conclusion Hprt1,GAPDH and Ppia are the most ideal reference genes for the screening of anti-HPIV-2 drugs and studying the interaction between HPIV-2 and LLC-MK2 in vitro.

关 键 词：副流感病毒2型 人 内参基因 稳定表达 实时聚合酶链反应 恒河猴肾细胞 

分 类 号：R186[医药卫生—流行病学]