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Tat抗微小隐孢子虫防御机制研究

发表时间:2017-09-15  浏览量:1726  下载量:263
全部作者: 周蕊,刘晓洁
作者单位: 武汉大学基础医学院
摘 要: 研究探讨Tat蛋白对miRNA介导的上皮细胞微小隐孢子虫感染的影响。通过实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)、Western blotting及荧光素酶活性检测等实验技术研究发现,使用含有HIV Tat的质粒转染细胞可以增强微小隐孢子虫的感染。并且发现在HIV Tat作用下微小隐孢子虫感染后抗病原生物相关因子诱导型一氧化氮合成酶(inducible nitric oxide synthase,iNOS)的表达量下降。在对其分子机制的研究中发现,Tat蛋白通过调节miR-27b的表达,从而影响KH型剪接调控蛋白(KH-type splicing regulatory protein,KSRP)的表达。KSRP作为一种RNA结合蛋白,可以与3'-非编码区(3'-UTR)区域中的ARE结构结合,调控iNOS mRNA的稳定性,从而影响上皮细胞抗病原生物防御机制,导致微小隐孢子虫感染加重。该分子机制的阐明也可以有助于解释艾滋病人更易感染其他机会性致病病原体这一普遍现象。
关 键 词: 医学寄生虫学;HIV Tat蛋白;诱导型一氧化氮合成酶(iNOS);KH型剪接调控蛋白(KSRP);抗微小隐孢子虫防御
Title: Investigation of Tat impairing anti-Cryptosporidium parvum defense mechanism
Author: ZHOU Rui, LIU Xiaojie
Organization: School of Basic Medical Sciences, Wuhan University
Abstract: The purposes of this paper are to investigate the effects of HIV Tat protein on miRNA-mediated epithelial anti-Cryptosporidium parvum defense. Using quantitative real-time PCR (qRT-PCR), Western blotting and luciferase assay, we demonstrated that HIV Tat decreased the expression of inducible nitric oxide synthase (iNOS) and induced epithelial expression of KH-type splicing regulatory protein (KSRP) (also known as KHSRP) through down regulation of selected miRNAs, such as miR-27b. As an RNA-binding protein, KSRP could recognize AU-rich element (ARE) within the 3'-untranslated region (3'-UTR) of iNOS mRNA and control their half-life time in the cytoplasm which impairs the defense of epithelial cell to C. parvum infection. Results from our studies could provide a rational basis for the implementation of new immunotherapeutic strategies for opportunistic infections in AIDS patients.
Key words: medical parasitology; HIV Tat protein; inducible nitric oxide synthase (iNOS); KH-type splicing regulatory protein (KSRP); anti-Cryptosporidium parvum defense
发表期数: 2017年9月第17期
引用格式: 周蕊,刘晓洁. Tat抗微小隐孢子虫防御机制研究[J]. 中国科技论文在线精品论文,2017,10(17):1942-1949.
 
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