摘要
目的研究A20蛋白与Toll样受体1(TLR1)和Toll样受体2(TLR2)信号通路产生炎症因子的关系,探讨A20蛋白在TLR1和TLR2信号通路中的分子机制,为细菌性感染疾病的抗炎和基因治疗提供基础理论依据。方法预先用Pam3CSK4处理的THP-1细胞,用Pam3CSK4再刺激,分析Pam3CSK4预处理对Pam3CSK4诱导的细胞因子产生的影响;RT-PCR和Western blot检测Pam3CSK4处理后TLR1和TLR2及My D88的表达水平;采用siRNA转染抑制A20表达,验证A20在Pam3CSK4诱导免疫耐受中的作用。结果 Pam3CSK4预处理可下调Pam3CSK4再刺激所诱导的细胞因子表达水平,包括IL-1β、TNF-α及IL-8,同时Pam3csk4预处理下调Pam3csk4所诱导的JNK、p38及NF-κB的信号传导;另外Pam3CSK4显著上调A20的表达水平,且有剂量及时间依赖性;A20过表达细胞中,Pam3csk4诱导的促炎症细胞因子的表达被逆转。结论 A20是诱导Pam3csk4耐受的重要调节因子,对Pam3csk4诱导的炎症反应具有负调控作用。
Objective To study the relationship between A20 and the induction of Pam3CSK4 tolerance,and to identify the mechanism of Pam3CSK4 tolerance induced by A20. Methods THP-1 cells,pretreatment with Pam3CSK4,were re-stimulated with Pam3CSK4. The effect of Pam3CSK4 pretreatment on cytokine production was induced by Pam3CSK4 stimulation,which was analyzed by RT-PCR. Western blot was used to detect the expression of TLR1 and TLR2 and My D88 after Pam3CSK4 treatment. SiRNA transfection was used to suppress the expression of A20 gene,which was used to determine the role of A20 protein in Pam3CSK4 induced immune tolerance. Results Pam3CSK4 pre-treatment down-regulated the expression levels of cytokines induced by Pam3csk4 re-stimulation,including IL-1β,TNF-α and IL-8. Pam3csk4down-regulated the signal transduction of JNK,p38 and NF-κB. Pam3CSK4 also significantly up-regulated A20 expression in a dose-and time-dependent manner. In THP-1 cells with A20 over-expression,the expression of pro-inflammatory cytokines was reversed. Conclusion A20 is an important negatively regulator for Pam3CSK4 induced immune tolerance.
作者
周李娜
胡锦跃
张平安
ZHOU Li - na HU Jin - yue ZHANG Ping - an(Department of Laboratory Science, The Peoples Hospital of Wuhan University, Wuhan Hubei 430060 , China Medical Research Center, Chang-sha Central Hospital, Changsha Hunan 410001, China.)
出处
《临床和实验医学杂志》
2017年第9期862-866,共5页
Journal of Clinical and Experimental Medicine
