| E3泛素连接酶Nrdp1和SOCS-1对布鲁氏菌诱导细胞凋亡的影响 |
| |
| 引用本文： | 江雅丽,陈创夫,车召堂,王震,李爽,张欢,张辉,郭飞.E3泛素连接酶Nrdp1和SOCS-1对布鲁氏菌诱导细胞凋亡的影响[J].中国畜牧兽医,2016,43(6):1393-1404. |
| |
| 作者姓名： | 江雅丽 陈创夫 车召堂 王震 李爽 张欢 张辉 郭飞 |
| |
| 作者单位： | 1. 石河子大学生命科学学院, 石河子 832000;2. 石河子大学动物科技学院, 石河子 832003;3. 石河子大学医学院, 石河子 832003 |
| |
| 基金项目： | 国家科技合作项目(2013DFA32380);国家科技支撑计划子课题(2013BAI05B05);国家自然科学基金(1572491、31502067、31260596);自治区研究生教育创新计划科研创新项目(XJGRI2015040) |
| |
| 摘 要： | 为了探讨E3泛素连接酶Nrdp1、SOCS-1基因在布鲁氏菌侵染巨噬细胞过程中对细胞凋亡的影响,构建Nrdp1、SOCS-1基因的干扰和过表达细胞模型(简称为pLL3.7-N1、pLL3.7-S1和pLEX-Nrdp1、pLEX-SOCS-1);羊种布鲁氏菌16M(简称16M) 侵染正常细胞RAW264.7及pLL3.7-N1、pLEX-Nrdp1、pLL3.7-S1、pLEX-SOCS-1组细胞,qRT-PCR检测Bax、Bcl-2、TNF-α的mRNA表达量,并通过流式细胞仪术检测细胞凋亡率。结果显示,试验成功构建并筛选出干扰和过表达Nrdp1、SOCS-1效果最好的pLL3.7-N1、pLEX-Nrdp1、pLL3.7-S1、pLEX-SOCS-1细胞模型;16M侵染各组细胞,与对照组相比,在不同的时间段pLL3.7-N1、pLL3.7-S1、pLEX-Nrdp1、pLEX-SOCS-1组细胞的Bcl-2和Bax的mRNA表达量差异显著(P < 0.05);pLL3.7-S1组细胞的TNF-α mRNA显著降低(P < 0.05),而pLEX-SOCS-1组显著升高(P < 0.05);pLEX-Nrdp1、pLEX-SOCS-1组细胞凋亡率显著升高(P < 0.05),而pLL3.7-S1组显著降低(P < 0.05)。研究结果表明,Nrdp1、SOCS-1基因与16M诱导的细胞凋亡密切相关,为16M胞内寄生机制的研究奠定了基础。
|
| 关 键 词： | 布鲁氏菌 Nrdp1 SOCS-1 实时荧光定量PCR 细胞凋亡 |
| 收稿时间： | 2015-11-08 |
Effect of E3 Ubiquitin Ligase Nrdp1 and SOCS-1 on Intracellular Survival and Macrophage Apoptosis Induced by Brucella |
| |
| JIANG Ya-li,CHEN Chuang-fu,CHE Zhao-tang,WANG Zhen,LI Shuang,ZHANG Huan,ZHANG Hui,GUO Fei.Effect of E3 Ubiquitin Ligase Nrdp1 and SOCS-1 on Intracellular Survival and Macrophage Apoptosis Induced by Brucella[J].China Animal Husbandry & Veterinary Medicine,2016,43(6):1393-1404. |
| |
| Authors: | JIANG Ya-li CHEN Chuang-fu CHE Zhao-tang WANG Zhen LI Shuang ZHANG Huan ZHANG Hui GUO Fei |
| |
| Institution: | 1. College of Life Technology, Shihezi University, Shihezi 832000, China;2. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;3. School of Medicine, Shihezi University, Shihezi 832003, China |
| |
| Abstract: | In order to investigate the effect of E3 ubiquitin ligase Nrdp1 and SOCS-1 genes on apoptosis during Brucella infection macrophages.The cell models of interference and over expression of Nrdp1 and SOCS-1 genes (pLL3.7-N1,pLL3.7-S1 and pLEX-Nrdp1,pLEX-SOCS-1) were constructed.Normal RAW264.7 cell,pLL3.7-N1,pLEX-Nrdp1,pLL3.7-S1 and pLEX-SOCS-1 group cells were infected with Brucella melitensis 16M (referred to 16M),the expression of Bax,Bcl-2,TNF-α genes were detected by qRT-PCR and apoptosis rate was detected by flow cytometry.pLL3.7-N1,pLEX-Nrdp1,pLL3.7-S1,pLEX-SOCS-1 cell model of the works best of interference and overexpression Nrdp1,SOCS-1 were successfully constructed and screened.After 16M infecting each group cells,compared with the control group,the expression of Bcl-2 and Bax mRNA in pLL3.7-N1,pLEX-Nrdp1,pLL3.7-S1 and pLEX-SOCS-1 groups were significantly different in different time periods (P < 0.05).The expression of TNF-α mRNA in pLL3.7-S1 group was significantly lower (P < 0.05),while the pLEX-SOCS-1 group was significantly higher (P < 0.05).Apoptosis rates in pLEX-Nrdp1,pLEX-SOCS-1 groups were significantly higher (P < 0.05),while pLL3.7-S1 group was lower (P < 0.05).The results showed that the Nrdp1 and SOCS-1 genes were closely related to apoptosis with 16M induction,the research laid the foundation for the study of 16M intracellular parasitism mechanisms. |
| |
| Keywords: | Brucella Nrdp1 SOCS-1 quantitative Real-time PCR apoptosis |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《中国畜牧兽医》浏览原始摘要信息 |
| 点击此处可从《中国畜牧兽医》下载免费的PDF全文 |
|
