E3泛素连接酶Nrdp1和SOCS-1对布鲁氏菌诱导细胞凋亡的影响

doi:10.16431/j.cnki.1671-7236.2016.06.001

›› 2016, Vol. 43 ›› Issue (6): 1393-1404.doi: 10.16431/j.cnki.1671-7236.2016.06.001

• 生物技术 • 下一篇

E3泛素连接酶Nrdp1和SOCS-1对布鲁氏菌诱导细胞凋亡的影响

江雅丽¹, 陈创夫², 车召堂¹, 王震², 李爽², 张欢², 张辉², 郭飞³

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

收稿日期:2015-11-08 出版日期:2016-06-20 发布日期:2016-07-11
通讯作者: 陈创夫, 郭飞 E-mail:ccf-xb@163.com;119783783@qq.com
作者简介:江雅丽(1989-),女,新疆伊犁人,硕士,研究方向:动物基因工程,E-mail:324622209@qq.com
基金资助:
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)

Effect of E3 Ubiquitin Ligase Nrdp1 and SOCS-1 on Intracellular Survival and Macrophage Apoptosis Induced by Brucella

JIANG Ya-li¹, CHEN Chuang-fu², CHE Zhao-tang¹, WANG Zhen², LI Shuang², ZHANG Huan², ZHANG Hui², GUO Fei³

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

Received:2015-11-08 Online:2016-06-20 Published:2016-07-11

摘要/Abstract

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

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.

Key words: Brucella; Nrdp1; SOCS-1; quantitative Real-time PCR; apoptosis

中图分类号:

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]. , 2016, 43(6): 1393-1404.

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]. , 2016, 43(6): 1393-1404.

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E3泛素连接酶Nrdp1和SOCS-1对布鲁氏菌诱导细胞凋亡的影响

Effect of E3 Ubiquitin Ligase Nrdp1 and SOCS-1 on Intracellular Survival and Macrophage Apoptosis Induced by Brucella

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