IL-4 induced MUC4 enhancement in respiratory epithelial cells in vitro is mediated through JAK-3 selective signaling.
IL-4 induced MUC4 enhancement in respiratory epithelial cells in vitro is mediated through JAK-3 selective signaling.
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DOI：
10.1186/1465-9921-7-39
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发表时间：
2006-03-21
影响因子：
5.8
通讯作者：
Sachdev, GP
中科院分区：
文献类型：
--
作者：
Damera, G;Xia, BY;Sachdev, GP
最近的研究发现MUC4粘蛋白是激活ErbB2的配体，ErbB2是一种受体酪氨酸激酶，可调节哮喘气道上皮损伤后上皮细胞的增殖。在这项研究中，我们利用NCI-H650细胞系研究了IL-4在调节MUC4表达中的潜在作用，IL-4是哮喘气道中持续存在的Th2炎症细胞因子之一。Real - time PCR检测IL-4对MUC4表达的浓度和时间依赖性。核运行实验进行了探索潜在的转录调节。采用MUC4 ASGP-2结构域特异性单克隆抗体进行Western blotting实验，分析质膜组分中MUC4糖蛋白水平。为了分析潜在的信号转导级联，在MUC4转录物分析之前，将IL-4处理的融合培养物与pan-JAK抑制剂、JAK-3选择性抑制剂或mek - 1,2 （MAPK）抑制剂分别在不同浓度下共孵育。使用单克隆p-STAT-6抗体进行western blotting检测相应转录因子的激活情况。MUC4水平以浓度和时间特异性的方式增加，在2.5 ng/ml和8 h时达到峰值表达。核运行实验显示转录增强。质膜部位MUC4糖蛋白水平相应升高。Pan-JAK抑制剂显示IL-4刺激MUC4水平显著降低，JAK3选择性抑制剂以浓度依赖性方式下调MUC4 mRNA表达。根据上述观察，在IL-4刺激后5分钟内检测到STAT-6的激活。使用MAPK抑制剂对MUC4水平没有影响。这些观察结果表明IL-4在气道上皮MUC4上调中的潜在作用。
Recent studies have identified MUC4 mucin as a ligand for activation of ErbB2, a receptor tyrosine kinase that modulates epithelial cell proliferation following epithelial damage in airways of asthmatics. In this study, we investigated the potential role of IL-4, one of the Th2 inflammatory cytokines persistent in asthmatic airways, in regulating MUC4 expression using a cell line NCI-H650. Real time PCR analysis was performed to determine concentration and time dependent effects of IL-4 upon MUC4 expression. Nuclear run on experiments were carried out to explore potential transcriptional modulation. Western blotting experiments using a monoclonal antibody specific to ASGP-2 domain of MUC4 were performed to analyze MUC4 glycoprotein levels in plasma membrane fractions. To analyze potential signal transduction cascades, IL-4 treated confluent cultures were co-incubated, separately with a pan-JAK inhibitor, a JAK-3 selective inhibitor or a MEK-1, 2 (MAPK) inhibitor at various concentrations before MUC4 transcript analysis. Corresponding transcription factor activation was tested by western blotting using a monoclonal p-STAT-6 antibody. MUC4 levels increased in a concentration and time specific fashion reaching peak expression at 2.5 ng/ml and 8 h. Nuclear run on experiments revealed transcriptional enhancement. Corresponding increases in MUC4 glycoprotein levels were observed in plasma membrane fractions. Pan-JAK inhibitor revealed marked reduction in IL-4 stimulated MUC4 levels and JAK3 selective inhibitor down-regulated MUC4 mRNA expression in a concentration-dependent fashion. In accordance with the above observations, STAT-6 activation was detected within 5 minutes of IL-4 stimulus. No effect in MUC4 levels was observed on using a MAPK inhibitor. These observations signify a potential role for IL-4 in MUC4 up-regulation in airway epithelia.
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影响因子：
6.1
作者：
AUDIE, JP;TETAERT, D;BOERSMA, A
通讯作者：
BOERSMA, A
影响因子：
3.2
作者：
Buisine, MP;Devisme, L;Porchet, N
通讯作者：
Porchet, N
DOI：
10.1164/ajrccm.164.supplement_2.2106066
发表时间：
2001-11-15
影响因子：
24.7
作者：
Fahy, JV
通讯作者：
Fahy, JV
影响因子：
4.4
作者：
Henderson, WR;Chi, EY;Maliszewski, CR
通讯作者：
Maliszewski, CR
影响因子：
3.8
作者：
Hunt, AE;Williams, LM;Foxwell, BMJ
通讯作者：
Foxwell, BMJ