敲低多房棘球蚴let-7-5p可抑制BALB/c小鼠腹腔巨噬细胞极化和虫体生长  

Knockdown of let-7-5p from Echinococcus multilocularis Inhibited the Peritoneal Macrophages Polarization and Worm Growth in BALB/c Mice

作　　者：王立群 吴易璇 蒲桂婷 曹珊菱 王得先 刘婷丽 李红 AMUDA Tharheer Oluwashola 郭小腊 殷宏[1,3] 骆学农 WANG Liqun;WU Yixuan;PU Guiting;CAO Shanling;WNAG Dexian;LIU Tingli;LI Hong;AMUDA Tharheer Oluwashola;GUO Xiaola;YIN Hong;LUO Xuenong(State Key Laboratory for Animal Disease Control and Prevention/Key Laboratory of Veterinary Parasitology of Gansu Province,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Life Science and Engineering College of Northwest University of Nationalities,Lanzhou 730030,China;Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonoses,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]中国农业科学院兰州兽医研究所动物疫病防控全国重点实验室/甘肃省动物寄生虫病重点实验室,兰州730046 [2]西北民族大学生命科学与工程学院,兰州730030 [3]扬州大学江苏省动物重要疫病与人兽共患病防控协同创新中心,扬州225009

出　　处：《畜牧兽医学报》2024年第6期2619-2628,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(32072889)。

摘　　要：旨在明确敲低多房棘球蚴emu-let-7-5p对小鼠腹腔巨噬细胞极化和虫体生长的影响。构建表达多房棘球蚴emu-let-7-5p海绵序列的重组腺相关病毒载体,并在293T细胞上验证其抑制效果和特异性。70只清洁级BALB/c小鼠分别经尾静脉注射AAV8-si-let-7-5p、AAV8-control和PBS后两周,每组随机选取2只小鼠分别用Western blot和qRT-PCR分析肝组织中GFP、let-7-5p及其靶基因C/EBP δ的表达。随后,所有小鼠每只腹腔接种1 000个原头蚴,3个月后qRT-PCR检测腹腔巨噬细胞emu-let-7-5p、C/EBP δ、M1型(IL-1β、iNOS和IL-6)和M2型(Arg-1、IL-4和IL-10)相关分子的转录水平,并统计各组小鼠多房棘球蚴的包囊重量和原头蚴数量。结果表明,注射后第15天,AAV8-si-let-7-5p成功靶向小鼠肝脏,并引起emu-let-7-5p的敲低和C/EBP δ的上调表达。感染后3个月,AAV8-si-let-7-5p组小鼠腹腔巨噬细胞emu-let-7-5p和M2型相关分子Arg-1均显著下调,而C/EBP δ和M1型相关分子IL-1β和iNOS均显著上调。此外,AAV8-si-let-7-5p组IL-4和IL-6下调表达,而IL-10上调表达,且小鼠肝脏包囊重量和原头蚴数量显著低于对照组。敲低多房棘球蚴感染小鼠体内emu-let-7-5p,可促进C/EBP δ的表达,进而抑制M2型极化和虫体生长。The purpose of this study was to determine the effects of knockdown of Echinococcus multilocularis emu-let-7-5p on mouse peritoneal macrophages polarization and worm growth.A recombinant adeno-associated virus vector expressing emu-let-7-5p sponge sequence of E.multilocularis was constructed,and its inhibitory effect and specificity were validated in 293T cells.Seventy clean-grade BALB/c mice were injected intravenously(tail vein)with AAV8-si-let-7-5p,AAV8-control,and PBS,respectively.After two weeks,two mice in each group were randomly selected,and the expression of GFP,emu-let-7-5p and its target C/EBPδin liver were detected by Western blot and qRT-PCR.At 3 months after intraperitoneal inoculation of 1000 protoscolexs in each mouse,the transcriptional levels of emu-let-7-5p,C/EBPδ,M1 type(IL-1β,iNOS and IL-6)and M2 type(Arg-1,IL-4 and IL-10)related molecules in peritoneal macrophages were detected by qRT-PCR,and cysts weight and protoscolexs number of E.multilocularis in each group were analyzed.The results showed that,on the 15th day after injection,AAV8-si-let-7-5p successfully targeted the mouse liver and caused knockdown of emu-let-7-5p and up-regulation of C/EBPδ.At 3 months after infection,the expressions of emu-let-7-5p and M2 related molecule(Arg-1)in peritoneal macrophages were significantly down-regulated in AAV8-si-let-7-5p group,while C/EBPδand M1 related molecules(IL-1βand iNOS)were significantly up-regulated.In addition,in AAV8-si-let-7-5p group,the expressions of IL-4 and IL-6 were down-regulated,while the expression of IL-10 was up-regulated,and cysts weight and protoscolexs number in AAV8-si-let-7-5p group were significantly lower than those in the control group.The above results suggested that knockdown of emu-let-7-5p in mice infected with E.multilocularis can promote the expression of C/EPBδ,and thus inhibit M2 macrophage polarization and worm growth.

关 键 词：多房棘球蚴 emu-let-7-5p 腹腔巨噬细胞极化 虫体生长 

分 类 号：S852.734[农业科学—基础兽医学]