Cytokine upregulation of proteinase-activated-receptors 2 and 4 expression mediated by p38 MAP kinase and inhibitory kappa B kinase β in human endothelial cells
Cytokine upregulation of proteinase-activated-receptors 2 and 4 expression mediated by p38 MAP kinase and inhibitory kappa B kinase β in human endothelial cells
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DOI：
10.1038/sj.bjp.0707150
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发表时间：
2007-04-01
影响因子：
7.3
通讯作者：
Plevin, R.
中科院分区：
文献类型：
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作者：
Ritchie, E.;Saka, M.;Plevin, R.
背景和目的：蛋白酶激活受体2（PAR2）的上调是许多疾病状态的一个因素，因此我们检查了与受体表达相关的信号通路。实验方法：我们研究了肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、胰蛋白酶和PAR2激活肽2-呋喃酰（2f）-LIGKV-OH对两者的影响人脐静脉内皮细胞 (HUVEC) 中 PAR2 的 mRNA 和功能表达。评估了特定化学抑制剂和显性失活腺病毒构建体对丝裂原激活蛋白激酶 (MAPK) 级联和核因子 kappa B (NF-kappa B) 信号通路的影响。方法包括半定量和定量RT-PCR、[H-3]磷酸肌醇(IP)积累和Ca2+依赖性荧光。主要结果：上述激动剂诱导PAR2的mRNA和功能表达； TNF α 治疗后，PAR4 mRNA 也增加，但 PAR1 或 PAR-3 的 mRNA 没有增加。抑制 p38 MAP 激酶会降低 PAR2 和 PAR4 的表达，而抑制 MEK1/ERK/JNK 则没有效果。 PAR4 的表达也观察到对 p38 MAP 激酶的类似依赖性。 TNF α 诱导的 PAR2 增强刺激了 [ H-3]-肌醇磷酸盐积累 (IP)，并且 Ca2+ 信号传导在 SB203580 预处理后被消除。感染编码显性失活 IKK beta (Ad.IKK beta(+/-)) 和较小程度的显性失活 IKK α (Ad.IKK alpha(+/-)) 的腺病毒，显着降低对照和 IL-1 β 诱导的 PAR2 和 PAR4 mRNA 表达，并增强 PAR2 刺激的 IP 积累和 Ca2+ 动员。结论和含义：这些数据首次揭示了参与上调的信号事件促炎挑战期间的 PAR2 和 PAR4。
Background and purpose: Up-regulation of proteinase-activated receptor-2 ( PAR2) is a factor in a number of disease states and we have therefore examined the signalling pathways involved in the expression of the receptor.Experimental approach: We investigated the effects of tumour necrosis factor-alpha ( TNF-alpha), interleukin-1 beta ( IL-1 beta), trypsin and the PAR2 activating peptide, 2-furoyl(2f)-LIGKV-OH on both mRNA and functional expression of PAR2 in human umbilical vein endothelial cells ( HUVECs). The effect of specific chemical inhibitors and dominant negative adenovirus constructs of the mitogen-activated protein kinase ( MAPK) cascade and the nuclear factor kappa B ( NF-kappa B) signalling pathway was assessed. Methods included semi-quantitative and quantitative RT-PCR, [ H-3] inositol phosphate ( IP) accumulation and Ca2+-dependent fluorescence.Key results: The above agonists induced both mRNA and functional expression of PAR2; PAR4 mRNA, but not that for PAR1 or PAR-3, also increased following TNF alpha treatment. Inhibition of p38 MAP kinase reduced PAR2 and PAR4 expression, whilst inhibition of MEK1/ERK/JNK was without effect. A similar dependency upon p38 MAP kinase was observed for the expression of PAR4. TNF alpha-induced enhancement of PAR2 stimulated [ H-3]-inositol phosphate accumulation ( IP) and Ca2+ signalling was abolished following SB203580 pre-treatment. Infection with adenovirus encoding dominant-negative IKK beta ( Ad.IKK beta(+/-)) and to a lesser extent dominant-negative IKK alpha ( Ad.IKK alpha(+/-)), substantially reduced both control and IL-1 beta-induced expression of both PAR2 and PAR4 mRNA and enhancement of PAR2-stimulated IP accumulation and Ca2+ mobilisation.Conclusions and implications: These data reveal for the first time the signalling events involved in the upregulation of both PAR2 and PAR4 during pro-inflammatory challenge.