程序性细胞死亡因子4基因在体干扰大鼠模型的构建  

Construction of the knockdown rat model of programmed cell death 4 in vivo

作　　者：钟波[1] 侯伟[1] 吕社民 王慧渊 耿妍[1] 杨旭东 薛丽[3] ZHONG Bo;HOU Wei;LV She-min;WANG Hui-yuan;GENG Yan;YANG Xu-dong;XUE Li(Department of Pediatrics, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004;Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an 710061;Clinical Laboratory, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, China)

机构地区：[1]西安交通大学第二附属医院小儿内科,陕西西安710004 [2]西安交通大学医学部基础医学院生物化学与分子生物学系,陕西西安710061 [3]西安交通大学第二附属医院检验科,陕西西安710004

出　　处：《西安交通大学学报（医学版）》2019年第5期789-793,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81800017);陕西省自然科学基础研究计划(No.2017JQ8045);西安交通大学基本科研业务费自由探索与创新项目(No.1191329843);西安交通大学第二附属医院院基金[No.YJ(QN)201504]资助项目~~

摘　　要：目的构建大鼠程序性细胞死亡因子4(programmed cell death 4,Pdcd4)基因在体干扰模型,为进一步研究该基因的功能提供条件。方法通过给予抗原诱导肺部炎症模型大鼠,鼻腔滴入Pdcd4的干扰质粒,以下调其体内Pdcd4的表达。收集大鼠支气管肺泡灌洗液细胞,提取总RNA,RT-qPCR检测Pdcd4基因mRNA的表达水平。结果在模型大鼠鼻腔滴入Pdcd4的干扰质粒,大鼠支气管肺泡灌洗液细胞中的Pdcd4的mRNA表达明显下调。结论成功构建了Pdcd4基因的在体干扰大鼠模型,为进一步研究Pdcd4基因的功能和作用机制奠定基础。Objective To construct the rat Pdcd4 knockdown model of programmed cell death 4 (Pdcd 4) in vivo for further study of the gene s functions.Methods E3 rats were intranasally instilled with saline or plasmids during antigen induced pulmonary inflammation model induction.The rats were anaesthetized and killed on day 21.The lungs in each group were lavaged by instillation and withdrawal of 2 mL ice-cold PBS for three times through the tracheal route, and the bronchoalveolar lavage fluid (BALF) was collected.After centrifugation, the BALF cell pellet was used to isolate the total RNA.And the mRNA level of Pdcd4 was assessed by real-time quantitative PCR.Results Pdcd4 expression decreased significantly in BALF cells at the mRNA level after Pdcd4 RNAi in vivo .Conclusion Rat Pdcd4 knockdown model in vivo was constructed successfully, which can lay foundation for further studies on the functions and mechanism of Pdcd4 gene.

关 键 词：程序性细胞死亡因子4 基因模型构建 在体干扰 支气管哮喘 

分 类 号：Q789[生物学—分子生物学]