类风湿关节炎中白三烯B4诱导TNF-α和IL-1β的表达  

LTB4 Induce Expression of TNF-α and IL-1β in Rheumatoid Arthritis

作　　者：陈占昆[1] 吕厚山[1] 林剑浩[1] 许少华[1] 蒋东芳[1] 

机构地区：[1]北京大学人民医院关节病研究中心,北京100044

出　　处：《中国生物化学与分子生物学报》2006年第5期415-421,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家重点基础研究发展项目(973计划)(No.2002CB513007)~~

摘　　要：为了探讨类风湿关节炎的发病过程中白三烯B4(leukotriene,LTB4)对TNFα和IL1β表达的影响.加入外源性LTB4或者在LIT存在的情况下,加入苯丁抑制素(bestatin,LTA4水解酶抑制剂)和MK886(5脂氧合酶激动蛋白抑制剂)后,采用实时PCR和酶联免疫吸附分析法来检测原代培养的类风湿滑膜细胞及培养上清液中TNFα和IL1β在mRNA及蛋白水平的表达.外源性的LTB410-8molL使TNFα和IL1βmRNA水平表达分别增加了14倍和1倍,蛋白水平分别增加了3倍.加入LIT刺激内源性的LTB4增加了14倍后,使TNFα和IL1βmRNA水平表达分别增加了145倍和12倍,蛋白水平分别增加了3倍.在LIT存在的情况下,MK88610μmolL使LTB4合成降低了62%(P<0.0001),使TNFα和IL1βmRNA水平表达分别降低了66%(P<0.05)和71%(P<0.001),它们的蛋白水平分别降低了75%和70%(P<0.01).100μgml苯丁抑制素使LTB4合成降低了78%(P<0.0001),使TNFα和IL1βmRNA水平表达分别降低了86%(P<0.001)和79%(P<0.01),它们的蛋白水平分别降低了84%和76%(P<0.05).在类风湿关节炎中,LTB4诱导TNFα和IL1β的表达.这一结果为类风湿关节炎发病机制进一步探讨提供了一条新思路.To investigate the effect of leukotriene B4 （LTB4） on expressions of TNF-α and IL-1β in pathogenesis of rheumatoid arthritis （RA）, primary cultured synovial cells from RA patients were treated with exogenous LTB4, LIT or MK-886 （inhibitor of 5-lipoxygenase activating protein） and Bestatin （inhibitor of leukotriene A4 hydrolase） in the presence of LIT （lipopolysaccharid and ionomyein and thapsigargin） respectively, expressions of TNF-α and IL-β were detected at mRNA level by real time PCR and their protein production by ELISA. LTB4 （10^-9 mol/L～10^-8 mol/L） was shown to induce dose-dependent increase of mRNA and protein expression of TNF-α. In contrast, IL-1β was elevated dose-independently at both mRNA and protein level. 14-fold endogenous product of LTB4 by LIT significantly increased mRNA the expressions of TNF-α （145 times） and IL-1β （12 times）, while their protein production were increased 3 times, respectively. LIT-treated synoviocyte with addition of MK-886 （1 μmol/L- 10 μmol/L） was inhibited to secrete LTB4 dose-dependently, following the markedly down-regulated expressions of TNF-α and IL-1β both at mRNA and protein level. Bestatin （100 μg/ml） could also remarkably reduced LTB4 about 78% （P 〈 0.000 1） and diminished LTB4-induced mRNA expression of TNF-α （86%） and IL-1β （79%）, paralleled by decreased protein products about 84% and 76% respectively. The data support the view that expressions of TNF-α and IL-1β could be induced by LTB4, which provided a new clue to investigate extensively pathogenic mechanism of rheumatoid arthritis.

关 键 词：白三烯B4 肿瘤坏死因子-Α 白细胞介素-1Β 类风湿关节炎 

分 类 号：R593.22[医药卫生—内科学]