IRAK-M在内毒素诱导Kupffer细胞耐受中的表达及其意义  

Expression of IRAK-M in endotoxin-tolerant Kupffer cells and its significance

作　　者：李旭宏[1] 陈先锋[2] 游海波[2] 刘海忠[2] 刘作金[2] 龚建平[2] 

机构地区：[1]重庆三峡中心医院肝胆外科,重庆市404000 [2]重庆医科大学附属第二医院肝胆外科,重庆市400010

出　　处：《世界华人消化杂志》2009年第7期652-656,共5页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30471696;No.30500473~~

摘　　要：目的:观察内毒素耐受状态下Kupffer细胞中白介素-1受体相关激酶-M(IRAK-M)的表达并探讨其在Kupffer细胞内毒素耐受形成中的作用.方法:通过脂多糖(LPS,10μg/L)预处理建立内毒素耐受Kupffer细胞模型,并与对照组细胞进行比较;在LPS(100μg/L)刺激后不同时点,用酶联免疫吸附试验(ELISA)检测细胞培养液中TNF-α水平,逆转录聚合酶链反应(RT-PCR)检测细胞中TNF-α和IRAK-M的mRNA表达,TransAM NF-κB Kit检测细胞中NF-κB活性,蛋白印迹法(Western blot)检测细胞中IRAK-M蛋白表达.结果:LPS刺激在两组细胞中均引起TNF-α释放增加,TNF-αmRNA表达以及NF-κB活性增强,但耐受组细胞的三种指标明显低于对照组(P<0.05);对照组细胞在LPS刺激24h后才能检测出IRAK-M mRNA的表达,而耐受组细胞在LPS刺激前已可检测到IRAK-M的基因表达,且该表达在LPS刺激后迅速增强,于6h达高峰,24h时仍明显高于对照组(P<0.05);对照组细胞在LPS刺激24h后才有微弱的IRAK-M蛋白表达,而耐受组细胞在LPS刺激前已有IRAK-M的蛋白表达,并随LPS刺激进一步上调,两组有显著差异(P<0.01).结论:内毒素耐受状态下,Kupffer细胞中的IRAK-M表达明显增强,IRAK-M可能在Kupffer细胞内毒素耐受形成中起重要作用.AIM： To observe the expression of IRAK-M in endotoxin-tolerant Kupffer cells and to explore its role in endotoxin tolerance of Kupffer cells.METHODS： The model of endotoxin-tolerant Kupffer cells was established by lipopolysaccharide （LPS, 10 μg/L） pretreatment, and was compared with control cells. At different time points after LPS （100 μg/L） stimulation, the TNF-α level in culture medium was measured by ELISA. The mRNA expression of TNF-α and IRAK-M in Kupffer cells was detected by RT-PCR. TransAM NF-KB Kit was used to examine the activation of NF-KB. The expression of IRAK-M protein in Kupffer cells was detected by Western blot.RESULTS： In both groups, LPS stimulation increased TNF-α level in culture medium, and enhanced TNF-α mRNA expression and NF-α：B activation in Kupffer cells, but these parameters were significantly lower in tolerant group than in control group （P 〈 0.05）. In control group, the expression of IRAK-M mRNA could not be detected until 24 h after LPS stimulation. How- ever, this expression was slight in tolerant group before LPS stimulation and was rapidly upregulated after stimulation, peaking at 6 h and was still stronger than the expression in control group at 24 h （P 〈 0.05）. In addition, there was no IRAK-M protein expression in control group until 24 h after LPS stimulation, but this expres- sion could be detected in tolerant group before LPS stimulation and was up-regulated in a time- dependent manner. There was significant differ- ence between two groups （P 〈 0.01）.CONCLUSION： IRAK-M expression is sig- nificantly up-regulated in endotoxin-tolerant Kupffer cells and it may play an important role in endotoxin tolerance of Kupffer cells.

关 键 词：Kupffer细胞 内毒素耐受 白介素-1受体相关激酶-M 

分 类 号：R362[医药卫生—病理学]