国际麻醉学与复苏杂志   2023, Issue (10): 2-2
    
沙格列汀通过调控ROS/NLRP3信号通路减轻大鼠机械通气肺损伤
王佳佳, 蔚海霞, 马源, 李忠林, 秦伟伟, 马福国, 王明山, 孙立新1()
1.青岛市市立医院
Saxagliptin attenuates ventilator-induced lung injury in rats by regulating ROS/NLRP3 signaling pathway
 全文:
摘要：

目的 探索活性氧（reactive oxygen species, ROS）/NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor pyrin domain containing 3, NLRP3）信号通路在沙格列汀减轻大鼠机械通气肺损伤（ventilator-induced lung injury, VILI）中的作用。方法 6~8周龄SPF级健康雄性SD大鼠36只，体重240~300 g，按照随机数字表法分为3组（每组12只）：对照组（C组）、VILI组（V组）及VILI+沙格列汀组（S组）。大潮气量机械通气4 h复制大鼠VILI模型。各组气管插管前及V组、S组机械通气4 h后或C组气管插管自主呼吸4 h后留取股动脉血行动脉血气分析检测PaO2及葡萄糖水平。处死大鼠后采用ELISA法检测血清及肺泡灌洗液（bronchoalveolar lavage fluid, BALF）中IL-1β和IL-18浓度，测量肺组织湿/干重比（wet/dry ratio, W/D），观察肺组织病理学改变并行肺损伤评分。Western blot法测定NLRP3、ASC和caspase-1的蛋白质表达水平。采用DCFHDA荧光探针法测定肺组织ROS表达水平。结果 与C组相比，V组机械通气结束时葡萄糖水平升高，V组和S组机械通气结束时PaO2降低，W/D比值、肺损伤评分、血清及BALF中IL-1β及IL-18浓度升高（P0.01），肺组织NLRP3、ASC和caspase-1的蛋白质表达上调（P0.05），肺组织ROS表达增加；与V组相比，S组机械通气结束时PaO2升高，葡萄糖水平降低，W/D比值、肺损伤评分、血清及BALF中IL-1β及IL-18浓度降低（P0.01），肺组织NLRP3、ASC、caspase-1的蛋白质表达下调（P0.05），肺组织ROS表达减少。结论 沙格列汀通过抑制ROS/NLRP3信号通路在VILI中发挥保护作用。
关键词： 沙格列汀；机械通气肺损伤；活性氧；NLR家族，蛋白结构域包含蛋白3
Abstract:

Objective To explore the role of reactive oxygen species (ROS)/NOD-like receptor pyrin domain containing 3 (NLRP3) signaling pathway on alleviated ventilator-induced lung injury (VILI) by saxagliptin in rats. Methods Thirty-six SPF-grade healthy Sprague-Dawley rats aged 6-8 weeks, weighing 240-300 g, according to the random number table method, they were randomly divided into 3 groups (n=12): control (group C), VILI (group V) and VILI + saxagliptin (group S). Mechanical ventilation with high tidal volume for 4 hours to simulate a model of VILI in rats. Before endotracheal intubation, 4 hours after mechanical ventilation in Group V and S, or 4 hours after spontaneous respiration in Group C, femoral arterial blood was collected for arterial blood gas analysis to detect PaO2 and glucose levels. Rats were sacrificed, IL-1β and IL-18 concentrations in serum and BALF were determined by ELISA. The wet weight/dry weight ratio (W/D) , pathologic histology changes of lung tissue and lung injury scores were examined. Protein expression levels of NLRP3, ASC, and caspase-1 were detected by the Western blot. ROS expression levels in lung tissues were determined by DCFHDA fluorescence probe. Results Compared with group C, glucose levels were elevated at the end of mechanical ventilation in group V, PaO2 was decreased after ventilation, W/D ratio, lung injury score, IL-1β and IL-18 concentrations in serum and BALF were increased (P0.01), protein expression levels of NLRP3, ASC, and caspase-1 were up-regulated (P0.05), ROS expression was increased in the lung tissue in group V and group S. Compared with group V, PaO2 was elevated at the end of mechanical ventilation in group S, glucose levels, W/D ratio, lung injury score, IL-1β and IL-18 concentrations were decreased(P0.01), protein expression levels of NLRP3, ASC, and caspase-1 in lung tissues were downregulated (P0.05), ROS expression in the lung tissue was decreased in group S. Conclusion Saxagliptin reduces VILI, and the mechanism may be related to inhibition of ROS/NLRP3 signaling pathway.
Key words: Saxagliptin; Ventilator-induced lung injury; Reactive oxygen species; NLR Family, Pyrin Domain-Containing 3 Protein