【摘要】 目的：探讨重组人白细胞介素37（rhIL-37）干预对细颗粒物(PM_2.5)暴露诱导的人支气管上皮细胞细胞外基质沉积的治疗作用及机制。方法：通过不同浓度（6.25、12.5、25和50 mg/L）的PM_2.5干预正常人支气管上皮细胞16HBE建立气道损伤的细胞模型，用Western blot和细胞免疫荧光检测Wnt家族成员5a（Wnt5a）表达水平，选取Wnt5a表达量最高时的PM_2.5浓度为最佳干预浓度。实验分为3组：PM_2.5+siRNA组、PM_2.5+Wnt5a siRNA组及control+Wnt5a siRNA，观察纤连蛋白（fibronectin）、IL-6和IL-1β的表达水平及活性氧（ROS）的产生。此外，又将实验分为PM_2.5+rhIL-37组、PM_2.5组和control组，研究rhIL-37是否能够下调Wnt5a、fibronectin、IL-6和IL-1β的表达。结果：（1）给予不同浓度（6.25、12.5、25和50 mg/L）的PM_2.5干预16HBE细胞24 h后，Wnt5a的表达除PM_2.5（50 mg/L）组，均较control组显著增加（P<0.05）；在PM_2.5（12.5 mg/L）刺激下，Wnt5a表达最高（P<0.01）。（2）在PM_2.5（12.5mg/L）干预16HBE细胞24 h后，fibronectin表达达到峰值（P<0.01）。（3）敲减Wnt5a后fibronectin和核因子E2相关因子2（Nrf2）的表达，ROS的产生，以及IL-6和IL-1β的分泌均较control组显著减少（P<0.01）。（4）rhIL-37干预后再给予PM_2.5刺激，Wnt5a、fibronectin和Nrf2的表达，ROS的产生，以及IL-6和IL-1β的分泌均较PM_2.5组显著减少（P<0.05）。结论：在体外细胞模型中，PM_2.5可能通过Wnt5a调控炎症反应和氧化应激，从而诱导支气管上皮细胞外基质沉积；rhIL-37通过下调Wnt5a减轻炎症反应和氧化应激，从而减少细胞外基质沉积。更多还原

【Abstract】 AIM:To investigate the therapeutic effect of recombinant human interleukin-37 （rhIL-37） inter?vention on extracellular matrix deposition in human bronchial epithelial cells exposed to fine particulate matter (PM_2.5),and to explore the underlying mechanisms.METHODS:An airway injury cell model was established using normal human bronchial epithelial 16HBE cells treated with various concentrations （6.25,12.5,25 and 50 mg/L） of PM_2.5.Western blot and immunofluorescence were employed to assess the expression level of Wnt family member 5a （Wnt5a）.The con?centration of PM_2.5 that maximized Wnt5a expression was identified as the optimal concentration for further analysis.Thecells were divided into 3 groups,PM_2.5+siRNA,PM_2.5+Wnt5a siRNA and control+Wnt5a siRNA,to evaluate the expres?sion of fibronectin,the secretion of IL-6 and IL-1β,and the production of reactive oxygen species （ROS）.A subsequent set of experiments was organized into PM_2.5+rhIL-37,PM_2.5 and control groups to determine whether rhIL-37 down-regu?lates the expression of Wnt5a and fibronectin,and reduces the secretion of IL-6 and IL-1β.RESULTS:（1） After 24 h of exposure to various concentrations of PM_2.5,the expression of Wnt5a in 16HBE cells was significantly elevated,except at50 mg/L,compared with control group （P<0.05）.The highest Wnt5a expression was observed at 12.5 mg/L PM_2.5 （P<0.01）.（2） Fibronectin expression peaked after 24 h of exposure to PM_2.5 at 12.5 mg/L （P<0.01）.（3） Successful knock?down of Wnt5a resulted in decreased levels of fibronectin and nuclear factor E2-related factor （Nrf2）.Additionally,the production of ROS and the secretion of IL-6 and IL-1β were significantly reduced compared with control group （P<0.01）.（4） The administration of rhIL-37 before PM_2.5 exposure led to decreased levels of Wnt5a,fibronectin and Nrf2 compared with PM_2.5 group （P<0.05）.Furthermore,rhIL-37 treatment reduced the ROS production and the secretion of IL-6 and IL-1β （P<0.05）.CONCLUSION:In vitro cell models demonstrate that PM_2.5 can induce extracellular matrix deposition in bronchial epithelial cells,potentially through the modulation of inflammatory response and oxidative stress influenced by Wnt5a.Additionally,rhIL-37 appears to mitigate inflammatory response and oxidative stress by down-regulating Wnt5a.更多还原