慢病毒介导短发夹RNA干扰技术建立小鼠肺组织TRIF沉默模型  

Establishment of murine lung TRIF-silencing model by lentivirus-mediated short hairpin RNA interference

作　　者：谢军[1,2] 龙晓茹 任洛[1,2] 刘恩梅[1,2] 谢晓虹[1,2] XIE Jun;LONG Xiao-ru;REN Luo;LIU En-mei;XIE Xiao-hong(Liangjiang Department of Respiratory Medicine,Children′s Hospital of Chongqing Medical University,Chongqing 400014,China;Ministry of Education Key Laboratory of Child Development and Disorders/National Clinical Research Center for Child Health and Disorders/China International Science and Technology Cooperation Base of Child Development and Critical Disorders/Chongqing Key Laboratory of Pediatrics,Chongqing 400014,China;Liangjiang Department of Infectious Disease,Children′s Hospital of Chongqing Medical University,Chongqing 400014,China)

机构地区：[1]重庆医科大学附属儿童医院两江呼吸科,重庆市400014 [2]重庆医科大学附属儿童医院儿童发育疾病研究教育部重点实验室、国家儿童健康与疾病临床医学研究中心、儿童发育重大疾病国家国际科技合作基地、儿科学重庆市重点实验室,重庆市400014 [3]重庆医科大学附属儿童医院两江感染科,重庆市400014

出　　处：《广西医学》2021年第14期1704-1707,共4页Guangxi Medical Journal

基　　金：国家自然科学基金项目(81800009,81300007)。

摘　　要：目的通过慢病毒介导的短发夹RNA(shRNA)干扰技术建立小鼠肺组织β干扰素Toll/白细胞介素1受体结构域衔接蛋白(TRIF)沉默模型。方法构建TRIF的siRNA干扰质粒及慢病毒载体。将18只Balb/c小鼠随机分为空白组、实验组、对照组,每组6只。分别将含TRIF-shRNA、对照慢病毒载体的病毒液经鼻滴入转染实验组、对照组小鼠的肺组织。第8天处死小鼠取肺组织检测绿色荧光蛋白表达情况,取肺脏、肝脏、脾脏、肾脏、心脏组织检测TRIF mRNA和蛋白的表达水平。结果实验组肺组织中可见明显绿色荧光蛋白表达,且肺组织中的TRIF mRNA和及蛋白表达水平均低于空白组及对照组(均P<0.05)。3组肝脏、脾脏、肾脏、心脏组织中的TRIF mRNA表达水平差异无统计学意义(均P>0.05)。结论使用滴鼻方法可有效建立肺组织局部TRIF沉默小鼠模型。Objective To establish a murine lung Toll/interleukin 1 receptor domain containing adaptor inducing interferonβ(TRIF)-silencing model by lentivirus-mediated short hairpin RNA(shRNA)interference.Methods siRNA interfered TRIF plasmid and lentiviral vector were constructed.Eighteen Balb/c mice were randomized to blank group,experimental group or control group,with six mice in each group.By nasal instillation,viral suspension containing TRIF-shRNA or control lentiviral vector was tansfected into murine lung tissues of the experimental and control groups respectively.The mice were sacrificed on the 8th day to harvest their lung tissues for detecting green fluorescent protein expression,and the lung,liver,spleen,kidney and heart tissues were removed to determine the expression levels of TRIF mRNA and protein.Results In the experimental group,obvious green fluorescent protein expression was observed,and lower lung tissue TRIF mRNA and protein expression levels were found as compared with those in the blank and control groups(all P<0.05).There was no statistically significant difference in expression level of TRIF mRNA in liver,spleen,kidney or heart tissues among the three groups(all P>0.05).Conclusion The murine lung local TRIF-silencing model can be successfully established by nasal instillation.

关 键 词：β干扰素Toll/白细胞介素1受体结构域衔接蛋白 慢病毒 干扰技术 肺组织 滴鼻 小鼠模型 

分 类 号：R392-33[医药卫生—免疫学]