外泌体包裹的miR-19b抑制EV71复制的影响
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摘要：
[目的]探讨外泌体包裹的miR-19b对EV71病毒复制的影响,为EV71病毒的miRNA防治药物的开发提供参考.[方法]将游离的miR-19b-cy3.5分子和转染miR-19b-cy3.5的HT29细胞分泌的外泌体分别处理T98G细胞,观察进入细胞的效率;分别提取未处理的、转染mimic-n.c.和转染miR-19b的HT29细胞分泌的外泌体,并给予T98G细胞孵育后,检测细胞内miR-19b的表达;分别以60 mg/L转染mimic-n.c.的HT29细胞分泌的外泌体、60 mg/L和40 mg/L转染miR-19b的HT29细胞分泌的外泌体孵育T98G细胞并感染EV71病毒,检测病毒RNA载量;将携带包裹的miR-19b-cy3.5的HT29细胞来源的外泌体,通过尾静脉注射小鼠检测miR-19b在小鼠各个器官中的表达.[结果]miR-19b-cy3.5能够通过外泌体的形式进入T98G细胞中,而游离的miR-19b-cy3.5无法进入细胞;转染miR-19b的HT29细胞分泌的外泌体在处理T98G细胞后,胞内miR-19b高表达,同时,外泌体处理后的T98G细胞中EV71病毒RNA复制量显著降低;动物实验中,外泌体包裹的miR-19b能够在小鼠的脑、肝及肺有效表达.[结论]外泌体包裹的miR-19b能够有效进入靶细胞中,并且可以抑制靶细胞中EV71病毒的复制;同时,外泌体包裹的miR-19b能够有效进入小鼠脑、肝以及肺等组织中并表达.
[Objective]The paper aims to examine exosome mediated miR-19b effect on the replication of Enterovirus 71 in T98G cells.[Methods]T98G cells were treated with miR-19b molecμles and exosomes secreted by miR-19b-cy3.5 transfected HT29 cells to determine their entry mode.T98G cells were respectively treated with exosomes secreted by normal HT29 cells,mimic-n.c.transfected HT29 cells and miR-19b transfected HT29 cells to detect miR-19b expression.60 mg/L exosomes secreted by mimic-n.c.transfected HT29 cells,60 mg/L and 40 mg/L exosomes secreted by miR-19b transfected HT29 cells were respectively co-cμltured with EV71 to detect the expression of viral RNA.[Results]The expression of miR-19b in mice was detected by intravenous injection of exosomes secreted by miR-19b-cy3.5 transfected HT29 cells.miR-19b mediated by exosomes coμld enter T98G cells.miR-19b was highly expressed in T98G cells treated with exosomes secreted by miR-19b transfected HT29 cells.The virus RNA significantly decreased in T98G cells treated with the exosomes secreted by miR-19b transfected HT29 cells.[Conclusion]miR-19b mediated by exosomes was expressed in the brain,liver and lungs of mice.exosomes mediated miR-19b was highly expressed in T98G cells,and concentration dependent inhibition of EV71 replication.It is expressed in mice brain,liver and lung.
作者：
李艳梅 程侠菊 王燕
Li Yanmei;Cheng Xiaju;Wang Yan(Institute of Biology and Medical Sciences,Soochow University,Suzhou 215123,China;Suzhou Municipal Hospital,Suzhou 215008,China)
机构地区：
苏州大学生物医学研究院 苏州市立医院
出处：
《betway官方app 学报：自然科学版》 CAS 北大核心 2023年第2期115-120,共6页
Journal of Henan Normal University(Natural Science Edition)
基金：
国家自然科学基金(81902913) 江苏高校优势学科建设工程资助项目(PAPD).
关键词：
miR-19b EV71 外泌体 MICRORNA
miR-19b EV71 exosomes microRNA
分类号：
R392 [医药卫生—免疫学]
