Chen Xu-lin, Xia Zhao-fan, Wei Duo, Ben Dao-feng, Wang Yong-jie, Deng Nian-qing
Department of Burns, the First Affiliated Hospital of Anhui Medical University, Hefei 230022, China.
Zhonghua Wai Ke Za Zhi. 2006 Apr 1;44(7):492-5.
To investigate the interaction between p38 mitogen-activated protein kinase signal transduction pathway and nuclear factor (NF)-kappaB/IkappaB system on the proinflammatory cytokines release after burn trauma.
Human monocyte line THP-1 were incubated with serum from eight healthy controls, burn sera, burn sera pretreatment with SB203580, and burn sera pretreatment with pyrrolidine dithiocarbamate (PDTC). After 24 hours incubation with serum, tumor necrosis factor (TNF)-alpha and interleukin-1beta (IL-1beta) levels in THP-1 culture supernatants were measured by ELISA. The activities of p38 MAPK and expressions of IkappaBalpha in THP-1 were measured by Western blot analysis. The EMSA method was used to characterize the binding activities of NF-kappaB and activating protein (AP)-1 in THP-1.
In comparison with normal controls, burn sera resulted in a significant higher level release of TNF-alpha and IL-1beta in THP-1 [(7.30 +/- 0.84) ng/ml vs (2.20 +/- 0.28) ng/ml, P < 0.05; (2.88 +/- 0.38) ng/ml vs (0.81 +/- 0.14) ng/ml, P < 0.05], which were significantly inhibited by pretreatment with SB203580 or PDTC. Burn sera showed increased activities of p38 MAPK and AP-1 in THP-1 (4728 +/- 582 vs 1291 +/- 163, P < 0.05; 946 +/- 137 vs 361 +/- 40, P < 0.05), which were abolished by pretreatment with SB203580 but not PDTC. The expression of IkappaBalpha in THP-1 incubated with burn sera was significantly decreased than those incubated with control sera (1211 +/- 115 vs 2658 +/- 318, P < 0.05), which were abolished by pretreatment with PDTC but not SB203580. Burn sera also leaded to an increased activity of NF-kappaB in THP-1 (1636 +/- 170 vs 317 +/- 32, P < 0.05), which were abolished by pretreatment with PDTC but not SB203580.
There are no direct interaction between p38 MAPK signal transduction pathway and NF-kappaB/IkappaB pathway. These two pathways, which regulate the production of TNF-alpha and IL-1beta in monocyte following burn trauma, are parallel and independent.
探讨p38丝裂原活化蛋白激酶信号转导通路与核因子（NF）-κB/IκB系统在烧伤后促炎细胞因子释放中的相互作用。
将人单核细胞系THP-1与来自8名健康对照者的血清、烧伤血清、用SB203580预处理的烧伤血清以及用吡咯烷二硫代氨基甲酸盐（PDTC）预处理的烧伤血清一起孵育。与血清孵育24小时后，通过酶联免疫吸附测定法（ELISA）检测THP-1培养上清液中肿瘤坏死因子（TNF）-α和白细胞介素-1β（IL-1β）的水平。通过蛋白质免疫印迹分析检测THP-1中p38丝裂原活化蛋白激酶（MAPK）的活性和IκBα的表达。采用电泳迁移率变动分析（EMSA）方法鉴定THP-1中NF-κB和活化蛋白（AP）-1的结合活性。
与正常对照相比，烧伤血清导致THP-1中TNF-α和IL-1β的释放水平显著升高[（7.30±0.84）ng/ml对（2.20±0.28）ng/ml，P<0.05；（2.88±0.38）ng/ml对（0.81±0.14）ng/ml，P<0.05]，而用SB203580或PDTC预处理可显著抑制这种升高。烧伤血清使THP-1中p38 MAPK和AP-1的活性增加（分别为4728±582对1291±163，P<0.05；946±137对361±40，P<0.05），用SB203580预处理可消除这种增加，但PDTC预处理不能。与对照血清孵育的THP-1相比，与烧伤血清孵育的THP-1中IκBα的表达显著降低（1211±115对2658±318，P<0.05），用PDTC预处理可消除这种降低，但SB203580预处理不能。烧伤血清还导致THP-1中NF-κB的活性增加（1636±170对317±32，P<0.05），用PDTC预处理可消除这种增加，但SB20^3580预处理不能。
p38 MAPK信号转导通路与NF-κB/IκB通路之间不存在直接相互作用。这两条通路在烧伤后调节单核细胞中TNF-α和IL-1β的产生，它们是平行且独立的。
