Li Sheng-wei, Liu Zuo-jin, Liu Chang-an, Li Xu-hong, You Hai-bo, Chen Xian-feng, Gong Jian-ping
Department of Hepatobiliary Surgery, Second Affiliated Hospital of Chongqing of Medical Sciences University, Chongqing 400010, China.
Zhonghua Gan Zang Bing Za Zhi. 2006 Feb;14(2):97-100.
To explore the mechanism of endotoxin tolerance (ET) through observing the expression of interleukin 1 receptor associated kinase-4 (IRAK-4) during endotoxin tolerance development in Kupffer cells (KCs).
Isolated KCs of Balb/c mouse were divided into two groups: the non-endotoxin tolerance (NET) group and the endotoxin tolerance (ET) group, which were pretreated with 10 ng/ml lipopolysaccharide (LPS) for 24 h. Then, the two groups were treated with 100 ng/ml LPS. The expressions of IRAK-4 gene and protein level were determined by RT-PCR and Western blot. The activities of NF-kappaB of KCs and the TNFalpha level were estimated by ELISA at 0 h, 1 h, 3 h, 6 h and 12 h after LPS stimulation.
The ultimate level of IRAK-4, the activities of NF-kappaB and the TNFalpha level were evidently lower in the ET group than those in the NET group (t = 12.4, 17.4 and 138.9 respectively, P<0.01).
Pretreatment with LPS on KCs could induce endotoxin tolerance of KCs and inhibition of IRAK-4 expression may be one of the reasons for its development.
通过观察库普弗细胞（KCs）内毒素耐受（ET）形成过程中白细胞介素1受体相关激酶4（IRAK-4）的表达，探讨内毒素耐受的机制。
将Balb/c小鼠分离的KCs分为两组：非内毒素耐受（NET）组和内毒素耐受（ET）组，两组均用10 ng/ml脂多糖（LPS）预处理24小时。然后，两组均用100 ng/ml LPS处理。采用RT-PCR和蛋白质印迹法检测IRAK-4基因表达和蛋白水平。在LPS刺激后0小时、1小时、3小时、6小时和12小时，通过ELISA法检测KCs中NF-κB的活性和TNFα水平。
ET组IRAK-4的最终水平、NF-κB的活性和TNFα水平明显低于NET组（t分别为12.4、17.4和138.9，P<0.01）。
KCs用LPS预处理可诱导KCs产生内毒素耐受，抑制IRAK-4表达可能是其形成的原因之一。
