Activated p38 MAPK in Peripheral Blood Monocytes of Steroid Resistant Asthmatics.
Activated p38 MAPK in Peripheral Blood Monocytes of Steroid Resistant Asthmatics.
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DOI：
10.1371/journal.pone.0141909
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Goleva E
中科院分区：
文献类型：
--
作者：
Li LB;Leung DY;Goleva E
类固醇抵抗是包括哮喘在内的慢性炎症性疾病治疗中的一个重大问题。需要可获得的生物标记物来识别类固醇耐药患者，以优化他们的治疗。这项研究检测了严重哮喘患者的皮质类固醇抵抗。24例预计1秒用力呼气量小于80%的哮喘患者，根据口服泼尼松治疗一周后肺功能的变化，被分为激素抵抗或激素敏感组。在口服泼尼松前采集患者的肝素化血液。用流式细胞仪检测全血中磷酸化丝裂原活化蛋白激酶(MAPK)(细胞外调节激酶(ERK)、p38和Jun激酶(JNK))的表达。免疫印迹法检测哮喘患者外周血单个核细胞(PBMC)中磷酸化p38MAPK和磷酸化丝裂原及应激激活蛋白激酶1(MSK1)的活性。实时荧光定量聚合酶链式反应检测地塞米松对激素耐药哮喘患者PBMC产生IL-8mRNA的抑制作用。流式细胞仪分析发现激素耐药组CD14+单核细胞p38磷酸化水平显著高于激素敏感组(p=0.014)，而两组哮喘患者CD14+细胞内ERK和JNK的磷酸化水平无明显差异。激素耐药组和激素敏感组的CD4+、CD8+T细胞、B细胞和NK细胞中p38、ERK、JNK的磷酸化水平无明显差异。Western印迹证实p38MAPK通路被激活，因为激素耐药哮喘患者PBMC裂解物中p38和msk1的磷酸化水平显著升高。P38抑制剂可显著增强激素抵抗型哮喘患者PBMC对内毒素诱导的IL-8mRNA的地塞米松抑制作用。这是首次在激素抵抗哮喘患者的血单核细胞中发现选择性激活p38MAPK通路的报道，提示p38和MSK1的磷酸化可以作为激素抵抗的血液生物标志物。
Steroid resistance is a significant problem in management of chronic inflammatory diseases, including asthma. Accessible biomarkers are needed to identify steroid resistant patients to optimize their treatment. This study examined corticosteroid resistance in severe asthma. 24 asthmatics with forced expiratory volume in one second of less then 80% predicted were classified as steroid resistant or steroid sensitive based on changes in their lung function following a week of treatment with oral prednisone. Heparinised blood was collected from patients prior to oral prednisone administration. Phosphorylated mitogen activated kinases (MAPK) (extracellular regulated kinase (ERK), p38 and jun kinase (JNK)) were analyzed in whole blood samples using flow cytometry. Activation of phospho-p38 MAPK and phospho-mitogen- and stress-activated protein kinase 1 (MSK1) in asthmatics’ peripheral blood mononuclear cells (PBMC) were confirmed by Western blot. Dexamethasone suppression of the LPS-induced IL-8 mRNA production by steroid resistant asthmatics PBMC in the presence of p38 and ERK inhibitors was evaluated by real time PCR. Flow cytometry analysis identified significantly stronger p38 phosphorylation in CD14+ monocytes from steroid resistant than steroid sensitive asthmatics (p = 0.014), whereas no difference was found in phosphorylation of ERK or JNK in CD14+ cells from these two groups of asthmatics. No difference in phosphorylated p38, ERK, JNK was detected in CD4+, CD8+ T cells, B cells and NK cells from steroid resistant vs. steroid sensitive asthmatics. P38 MAPK pathway activation was confirmed by Western blot, as significantly higher phospho-p38 and phospho-MSK1 levels were detected in the PBMC lysates from steroid resistant asthmatics. P38 inhibitor significantly enhanced DEX suppression of LPS-induced IL-8 mRNA by PBMC of steroid resistant asthmatics. This is the first report demonstrating selective p38 MAPK pathway activation in blood monocytes of steroid resistant asthmatics, suggesting that p38 and MSK1 phosphorylation can serve as blood biomarkers of steroid resistance.