Blau Hannah, Klein Keren, Shalit Itamar, Halperin Drora, Fabian Ina
Schneider Children's Medical Center of Israel, 14 Kaplan Street, Petah Tikva, Israel 49202.
Am J Physiol Lung Cell Mol Physiol. 2007 Jan;292(1):L343-52. doi: 10.1152/ajplung.00030.2006. Epub 2006 Sep 29.
Cystic fibrosis (CF) is associated with severe neutrophilic airway inflammation. We showed that moxifloxacin (MXF) inhibits IL-8 and MAPK activation in monocytic and respiratory epithelial cells. Azithromycin (AZM) and ciprofloxacin (CIP) are used clinically in CF. Thus we now examined effects of MXF, CIP, and AZM directly on CF cells. IB3, a CF bronchial cell line, and corrected C38 cells were treated with TNF-alpha, IL-1beta, or LPS with or without 5-50 microg/ml MXF, CIP, or AZM. IL-6 and IL-8 secretion (ELISA), MAPKs ERK1/2, JNK, p38, and p65 NF-kappaB (Western blot) activation were measured. Baseline IL-6 was sixfold higher in IB3 than C38 cells but IL-8 was similar. TNF-alpha and IL-1beta increased IL-6 and IL-8 12- to 67-fold with higher levels in IB3 than C38 cells post-TNF-alpha (P < 0.05). Levels were unchanged following LPS. Baseline phosphorylated form of ERK1/2 (p-ERK1/2), JNK, and NF-kappaB p65 were higher in IB3 than C38 cells (5-, 1.4-, and 1.4-fold), and following TNF-alpha increased, as did the p-p38, by 1.6- to 2-fold. MXF (5-50 microg/ml) and CIP (50 microg/ml), but not AZM, suppressed IL-6 and IL-8 secretion by up to 69%. MXF inhibited TNF-alpha-stimulated MAPKs ERK1/2, 46-kDa JNK, and NF-kappaB up to 60%, 40%, and 40%, respectively. In contrast, MXF did not inhibit p38 activation, implying a highly selective pretranslational effect. In conclusion, TNF-alpha and IL-1beta induce an exaggerated inflammatory response in CF airway cells, inhibited by MXF more than by CIP or AZM. Clinical trials are recommended to assess efficacy in CF and other chronic lung diseases.
囊性纤维化(CF)与严重的中性粒细胞气道炎症相关。我们发现莫西沙星(MXF)可抑制单核细胞和呼吸道上皮细胞中白细胞介素-8(IL-8)和丝裂原活化蛋白激酶(MAPK)的激活。阿奇霉素(AZM)和环丙沙星(CIP)在临床上用于CF治疗。因此,我们现在直接研究了MXF、CIP和AZM对CF细胞的影响。用肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)或脂多糖(LPS)处理CF支气管细胞系IB3和校正后的C38细胞,同时加入或不加入5 - 50微克/毫升的MXF、CIP或AZM。检测IL-6和IL-8的分泌(酶联免疫吸附测定法)、MAPK的细胞外信号调节激酶1/2(ERK1/2)、应激活化蛋白激酶(JNK)、p38和核因子-κB p65(p65 NF-κB)的激活(蛋白质印迹法)。IB3细胞的基线IL-6水平比C38细胞高6倍,但IL-8水平相似。TNF-α和IL-1β使IL-6和IL-8增加12至67倍,TNF-α刺激后IB3细胞中的水平高于C38细胞(P < 0.05)。LPS刺激后水平无变化。IB3细胞中ERK1/2的磷酸化形式(p-ERK1/2)、JNK和NF-κB p65的基线水平高于C38细胞(分别为5倍、1.4倍和1.4倍),TNF-α刺激后p-p38也增加,增加了1.6至2倍。MXF(5 - 50微克/毫升)和CIP(50微克/毫升),但不是AZM,可将IL-6和IL-8的分泌抑制高达69%。MXF分别将TNF-α刺激的MAPK的ERK1/2、46千道尔顿的JNK和NF-κB抑制高达60%、40%和40%。相比之下,MXF不抑制p38的激活,这意味着其具有高度选择性的翻译前效应。总之,TNF-α和IL-1β在CF气道细胞中诱导过度的炎症反应,MXF对其的抑制作用大于CIP或AZM。建议进行临床试验以评估其在CF和其他慢性肺部疾病中的疗效。