| 胡程舟,张卓,陈文龙,等.干扰素调节因子4在风湿性心脏病中去泛素化作用的研究[J].安徽医药,2019,23(4):646-650. |
| 干扰素调节因子4在风湿性心脏病中去泛素化作用的研究 |
| The function of deubiquitin modification of IRF4 in rheumatic heart disease |
| 投稿时间：2017-12-29 |
| DOI： |
| 中文关键词: 风湿性心脏病 Th2细胞 干扰素调节因子4 泛素化 泛素特异性蛋白4 |
| 英文关键词: Rheumatic heart disease Th2 Cells Interferon regulatory factor-4 Ubiquitination Ubiquitin specific protein 4 |
| 基金项目: |
|
| 摘要点击次数: 1795 |
| 全文下载次数: 570 |
| 中文摘要: |
| 目的 探究风湿性心脏病病人干扰素调节因子4(IRF4)去泛素化与去泛素化酶USP4互相作用的分子机制,为风湿性心脏病提供新的治疗策略。方法 来自安徽医科大学第一附属医院2015年7月至2016年1月20例风湿性心脏病病人,均行体外循环下瓣膜置换手术,均顺利出院。10例健康对照来自于健康志愿者。分别抽取以上参与实验人员5 mL的外周血,经过细胞培养与转染；通过免疫共沉淀实验观察IRF4的DNA结合结构域以及和配体结合结构域是否可以和USP4互相结合；通过NI-NTA镍螯合树脂纯化实验观察IRF4蛋白针对48位和63位赖氨酸相关的去泛素化是否可以被USP4介导；应用荧光素酶检测技术观察白细胞介素(IL)-4是否可以在IRF4、活化T细胞核因子(NFATC2)和USP4共同促进下表达；利用基因沉默方法在人源Th2细胞中下调USP4。结果 下调USP4后Th2细胞相关细胞因子的转录水平也明显减少；利用流式细胞术检测经USP4抑制剂处理后的Th2细胞表达的IRF4和IL-4均减少；以流式细胞技术测得风湿性心脏病中IL-4表达增高,IRF4表达也增高。结论 风湿性心脏病病人血中IRF4的蛋白稳定表达可以通过USP4去泛素化的作用而实现；IL-4在IRF4表达增加协同NFATC2的情况下可增加表达。 |
| 英文摘要: |
| Objective To explore the molecular mechanism of the interaction between deubiquitinating enzyme USP4 and deubiquitination of interferon regulatory factor 4 (IRF4) in rheumatic heart disease patients,and to provide new tactics for treatment of rheumatic heart disease (RHD).Methods Twenty RHD patients were included in the study,who underwent valve replacement under cardiopulmonary bypass and were discharged smoothly from July 2015 to January 2016 in The First Affiliated Hospital of Anhui Medical University.Ten healthy controls were from healthy volunteers.The peripheral blood of 5 mL of the above participants was collected,cultured and transfected.The experiment of coimmunoprecipitation was carried out to observe if USP4 could be combined with IRF4 DNA binding domain and ligand binding domain (LBD).The experiment of NI-NTA Nickel chelating resin purification was made to observe if USP4 could mediate deubiquitination of IRF4 protein related to 48 lysine and 63 lysine.And luciferase detection technology was used to observe if the expression of interleukin-4 (IL-4) could be promoted by IRF4,nuclear factor of activated Tcell (NFATC2) and USP4.Gene silencing method was used to down-regulate USP4 in human Th2 cells.Results Down regulation of USP4 showed that the related cytokines transcriptional level of Th2 cells was significantly reduced.Flow cytometry showed that the levels of IRF4 and IL-4 expressed with Th2 cells handled by USP4 inhibitor both reduced,and the expressions of IRF4 and IL-4both increased in rheumatic heart disease patients.Conclusions Through deubiquitination,USP4 can stabilize the expression of IRF4 protein in rheumatic heart disease patients.The increase of IRF4 can raise the expression of IL-4 in coordination with NFATC2. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
