Dept of Pediatrics, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.
Division of Physiology, School of Nursing and Rehabilitation Sciences, Showa University, Yokohama, Japan.
Eur Respir J. 2014 Oct;44(4):1002-10. doi: 10.1183/09031936.00080913. Epub 2014 May 15.
Bacterial lipopolysaccharide (LPS) and interleukin (IL)-13 increase mucus secretion and inflammatory cytokine production in normal human bronchial epithelial (NHBE) cells. We evaluated the effect of club cell 10-kDa protein (CC10), an anti-inflammatory protein produced by epithelial cells, on mucus secretion, cell morphology and inflammatory cytokine production. NHBE cells were cultured at an air-liquid interface with CC10 or vehicle and exposed to LPS on day 14. Mucin MUC5AC, IL-8 and granulocyte-macrophage colony-stimulating factor were measured in cell supernatants. MUC5AC and IL-8 mRNA expression were measured by real-time PCR. Western blotting was used to evaluate nuclear factor (NF)-κB and extracellular signal-regulated kinase (ERK) activation. Cells were evaluated histologically. Additionally, NHBE cells were exposed to IL-13 and CC10 for 14 days, and secretion of the mucins MUC5AC and MUC5B was measured. MUC5AC secretion stimulated either by LPS or by IL-13 was attenuated by CC10 at 20 ng·mL(-1) (p<0.05). CC10 at 20 ng·mL(-1) also attenuated IL-8 secretion (p<0.05). MUC5AC and IL-8 mRNA expression were also decreased by CC10 (p<0.05). CC10 attenuated phosphorylation of NF-κB (p<0.05) and ERK1/2 (p<0.05). CC10 attenuates LPS-induced mucus secretion in airway cells, in part due to inhibition of NF-κB and ERK phosphorylation.
细菌脂多糖 (LPS) 和白细胞介素 (IL)-13 可增加正常人气道上皮 (NHBE) 细胞的黏液分泌和炎症细胞因子产生。我们评估了上皮细胞产生的抗炎蛋白——克拉细胞 10kDa 蛋白 (CC10) 对黏液分泌、细胞形态和炎症细胞因子产生的影响。NHBE 细胞在气-液界面培养,给予 CC10 或载体,并在第 14 天暴露于 LPS。在细胞上清液中测量粘蛋白 MUC5AC、IL-8 和粒细胞-巨噬细胞集落刺激因子。通过实时 PCR 测量 MUC5AC 和 IL-8 mRNA 表达。Western blot 用于评估核因子 (NF)-κB 和细胞外信号调节激酶 (ERK) 的激活。对细胞进行组织学评估。此外,NHBE 细胞暴露于 IL-13 和 CC10 14 天后,测量粘蛋白 MUC5AC 和 MUC5B 的分泌。LPS 或 IL-13 刺激的 MUC5AC 分泌被 20ng·mL(-1) 的 CC10 减弱 (p<0.05)。20ng·mL(-1) 的 CC10 也减弱了 IL-8 分泌 (p<0.05)。CC10 还降低了 MUC5AC 和 IL-8 mRNA 表达 (p<0.05)。CC10 减弱了 NF-κB (p<0.05) 和 ERK1/2 (p<0.05) 的磷酸化。CC10 减弱 LPS 诱导的气道细胞黏液分泌,部分原因是抑制 NF-κB 和 ERK 磷酸化。