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AIR对布鲁氏菌感染引发的炎症反应的影响
引用本文:徐亚芳,陈创夫,王浩,付强,史慧君,孙志华,张辉,郭飞.AIR对布鲁氏菌感染引发的炎症反应的影响[J].中国畜牧兽医,2016,43(6):1437-1445.
作者姓名:徐亚芳  陈创夫  王浩  付强  史慧君  孙志华  张辉  郭飞
作者单位:1. 石河子大学动物科技学院, 石河子 832003;2. 新疆农业大学动物医学学院, 乌鲁木齐 830000;3. 海南大学热带生物资源教育部重点实验室, 海口 570228;4. 石河子大学医学院, 石河子 832002
基金项目:国家国际科技合作项目(2013DFA32380);国家科技支撑计划子课题(2013BAI05B05);国家自然科学基金(31502067)
摘    要:试验旨在研究布鲁氏菌侵染宿主细胞过程中AIR对由布鲁氏菌感染引发的炎症反应的影响。本试验分别对膜融合蛋白Tecpr1中AIR结构域进行干扰(I-A)、过表达(O-A)、干扰后回补(OA-IA),建立羊种布鲁氏菌16M侵染细胞的模型。以二氯荧光素(DCFH-DA)为探针,利用激光共聚焦显微镜观察 ROS产生的变化及各组细胞线粒体分布的动态变化;通过qRT-PCR技术检测16M侵染宿主细胞引起NLRP3、 ASC和Caspase-1表达量的变化;通过ELISA方法检测IL-18、IL-1β和Caspase-1炎性因子表达量的变化。结果表明,16M能以时间依赖性方式诱导RAW264.7细胞产生ROS,I-A组和 O-A组线粒体异常集聚程度高;qRT-PCR及ELISA检测结果表明,16M侵染宿主细胞能够引起不同处理组细胞中炎性相关基因NLRP3、ASC和Caspase-1及炎性因子IL-18、IL-1β和Caspase-1表达量的变化。AIR缺失后,ROS 释放量发生变化,线粒体出现异常集聚,且AIR与炎性小体的活化、炎症反应的发生密切相关。

关 键 词:布鲁氏菌  AIR  ROS  线粒体  炎性小体  
收稿时间:2015-10-08

Effect of AIR on Inflammatory Reaction Infected by Brucella
XU Ya-fang,CHEN Chuang-fu,WANG Hao,FU Qiang,SHI Hui-jun,SUN Zhi-hua,ZHANG Hui,GUO Fei.Effect of AIR on Inflammatory Reaction Infected by Brucella[J].China Animal Husbandry & Veterinary Medicine,2016,43(6):1437-1445.
Authors:XU Ya-fang  CHEN Chuang-fu  WANG Hao  FU Qiang  SHI Hui-jun  SUN Zhi-hua  ZHANG Hui  GUO Fei
Institution:1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;2. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830000, China;3. Key Laboratory for Tropical Biological Resources, Ministry of Education, Hainan University, Haikou 570228, China;4. School of Medicine, Shihezi University, Shihezi 832002, China
Abstract:In order to investigate the effect of AIR on inflammatory reaction infected by Brucellamelitensis (16M), the AIR domain of Tecpr1 gene of murine macrophages RAW264.7 were knocked down (I-A), overexpressed (O-A) and reversed (OA-IA). Using the chlorine fluorescein (DCFH-DA) as a probe, we detected the variation of ROS production and mitochondria distribution by confocal laser scanning microscopy. We observed the expression changes of NLRP3, ASC and Caspase-1 by qRT-PCR and the expression changes of IL-18,IL-1β and Caspase-1 in host cells by ELISA. The results showed that 16M could stimulate RAW264.7 cells to produce ROS by time-dependent pathway, and I-A group and O-A group showed more abnormal accumulation of mitochondrial. The results of qRT-PCR and ELISA suggested that it had effect on the expression levels of NLRP3, ASC,Caspase-1 and IL-18, IL-1β and Caspase-1 in cells of different groups. Those results indicated that with AIR gene deletion, the release amount of ROS changed, mitochondrial clustered abnormally, and AIR was closely related to the activation of inflammasomes and induction of inflammatory reactions.
Keywords:Brucella  AIR  ROS  mitochondria  inflammasome  
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