作　　者： (陈诗皓）; (杜晓楠）; (李艳）; (吕喆）; (秦啸峰）; (孙英）; (王炜）;

机构地区： 首都医科大学基础医学院免疫学系,北京100069

出　　处： 《微生物学免疫学进展》 2017年第4期1-7,共7页

摘　　要： 目的探讨气道上皮细胞及固有免疫细胞经粉尘螨刺激后,白细胞介素37(interleukin 37,IL-37)对其产生细胞因子IL-6的影响。方法体外培养人肺泡上皮细胞系A549、小鼠肺上皮细胞系MLE-12、小鼠巨噬细胞系RAW264.7和原代小鼠固有淋巴样2型细胞(ILC2细胞),当细胞融合度达70%时,用IL-37预处理2 h,再给予粉尘螨粗提物刺激细胞,每种细胞均设PBS、IL-37及粉尘螨对照,并分别于不同时间点收集细胞沉淀和细胞培养液上清;用实时聚合酶链反应(real-time polymerase chain reaction,real-time PCR)法检测细胞因子IL-6在mRNA水平上的变化,用双抗体夹心ELISA检测细胞因子IL-6在蛋白水平的变化,用流式细胞术分选ILC2细胞并检测ILC2细胞表面IL-37受体的表达情况。结果粉尘螨粗提物可促进A549细胞、MLE-12细胞、RAW264.7细胞和ILC2细胞表达I L-6,且呈时间依赖的方式(P<0.05)。IL-37可抑制粉尘螨粗提物所诱导的IL-6在A549细胞、MLE-12细胞和RAW264.7细胞表达(P<0.05);但IL-37对粉尘螨粗提物刺激ILC2细胞分泌IL-6无明显抑制作用(P>0.05)。结论IL-37可抑制粉尘螨诱导的气道上皮细胞和巨噬细胞产生IL-6,并可通过负向调控下调气道炎症反应,为IL-37在哮喘治疗中的潜在应用提供了实验依据。 Objective This study aimed to explore effects of IL-37 on production of IL-6 by epithelial cells on airway and innate immune cells stimulated with Dermatophagoides farinae（D. farinae） extract. Methods A total of 4 cell lines including human alveolar epithelial cell line A549,murine lung epithelial cell line MLE-12,murine macrophage cell line RAW264.7and primary murine group 2 innate lymphoid cells（ILC2s） were cultured in vitro in the presence of IL-37 for 2 h when the cells reached to 70% confluence,then stimulated with D. farinae extract. Each of tested cell lines was set up PBS,IL-37,D.farinae extract group,respectively,as controls. Cell lysates and the supernatants of cell cultures were collected at different time points. The mRNA levels of IL-6 were detected by using real-time PCR. The protein levels of IL-6 were measured by a double antibodies sandwich ELISA. Flow cytometry was used to isolate ILC2 s and to detect the expression of IL-37 R on ILC2 s. Results D. farinae extract significantly enhanced IL-6 expression in A549,MLE-12,RAW264.7 and ILC2s（P0.05）,which was in a time-dependent mode（P0.05）. IL-37 significantly inhibited IL-6 expression induced by D. farinae extract in A549,MLE-12 and RAW264.7（P 0.05）,but IL-37 had no significant effect on secretion of IL-6 induced by D.farinae extract in murine ILC2s（P0.05）. Conclusion IL-37 may negatively regulate the inflammatory response in airway through inhibiting production of pro-inflammatory IL-6 in epithelial cells and macrophages. The study provided an experimental foundation for potential application of IL-37 on asthma treatment.

关 键 词： 白细胞介素 粉尘螨 白细胞介素 哮喘