西格列汀预处理对大鼠机械通气相关性肺损伤NLRP3/caspase-1/GSDMD信号通路的影响  

Effect of sitagliptin preconditioning on the NOD-like receptor 3/cysteine aspartic acid specific protease-1/gasdermin D signaling pathway in rats with mechanical ventilation associated lung injury

作　　者：马源[1] 代玉婷 王佳佳[1] 秦伟伟[1] 孙立新[1] 张晓[1] 韩伟[2] Ma Yuan;Dai Yuting;Wang Jiajia;Qin Weiwei;Sun Lixin;Zhang Xiao;Han Wei(Department of Anesthesiology,Qingdao Municipal Hospital,the Medical College of Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学附属青岛市市立医院麻醉科,青岛266071 [2]青岛大学附属青岛市市立医院呼吸与危重症科,青岛266071

出　　处：《国际麻醉学与复苏杂志》2023年第12期1265-1271,共7页International Journal of Anesthesiology and Resuscitation

摘　　要：目的探究西格列汀预处理对大鼠机械通气相关性肺损伤(ventilator-induced lung injury,VILI)NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor 3,NLRP3)/胱天蛋白酶-1(cysteine aspartic acid specific protease-1,caspase-1)/消皮素D(gasdermin D,GSDMD)信号通路的影响。方法36只6~8周龄SPF级健康雄性SD大鼠,按随机数字表法分为3组(每组12只):对照组(C组)、大潮气量模型组(V组)和大潮气量机械通气+西格列汀组(S组)。3组大鼠均行气管切开,C组大鼠保持自主呼吸4 h,V组、S组气管切开后行机械通气4 h。气管切开前1 h S组大鼠腹腔注射100 mg/kg西格列汀,C组、V组大鼠腹腔注射等容量生理盐水。分别于气管切开前及机械通气或自主呼吸4 h后检测大鼠动脉血PaO_(2)及葡萄糖水平。采血后处死大鼠,ELISA法检测大鼠血清及支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中IL-1β、IL-18浓度。观察肺组织大体形态,光镜下观察大鼠肺组织H-E染色病理学形态,行肺损伤评分。检测肺组织湿重/干重(wet/dry,W/D)。Western blot法检测大鼠肺组织NLRP3、GSDMD、凋亡相关斑点样蛋白(apoptosis-related speck protein,ASC)、caspase-1表达水平。结果C组大鼠肺泡形态结构大致正常,V组大鼠肺泡完整性破坏,肺泡断裂融合,肺泡间隔水肿,可见大量炎症细胞浸润,肺泡内可见出血。与C组比较,V组、S组大鼠肺组织表面呈现不同程度的暗红色,可见充血水肿;与V组比较,S组出血水肿减轻,可见少量散在出血点,肺间质水肿减轻,炎症细胞浸润减少,损伤程度减轻。与C组比较:V组和S组大鼠肺损伤评分,W/D,血清及BALF中IL-1β、IL-18浓度升高(P<0.05),肺组织NLRP3、GSDMD、ASC、caspase-1水平上调(P<0.05),机械通气4 h后PaO2降低(P<0.05);V组机械通气4 h后葡萄糖水平升高(P<0.05)。与V组比较,S组肺损伤评分,W/D,血清及BALF中IL-1β、IL-18浓度降低(P<0.05),肺组织NLRP3、GSDMD、ASC、caspase-1水平下�Objective To explore the effect of sitagliptin preconditioning on the NOD-like receptor 3(NLRP3)/cysteine aspartic acid specific protease-1(caspase-1)/gasdermin D(GSDMD)signaling pathway in rats with mechanical ventilation associated lung injury(VILI)models.Methods A total of 36 specific pathogen free(SPF)grade,healthy Sprague-Dawley(SD)male rats,aging 6-8 weeks were selected.According to the random number table method,they were divided into three groups(n=12):a control group(group C),a high tidal volume model group(group V),and a high tidal volume ventilation+sitagliptin group(group S).Tracheotomy was performed in all the three groups.Then,group C maintained autonomous respiration for 4 h,while group V and group S received mechanical ventilation for 4 h.Then,1 h before tracheotomy,rats in group S were intraperitoneally injected with 100 mg/kg sitagliptin,while those in group C and group V were intraperitoneally injected the same volume of normal saline.The partial pressure of oxygen(PaO_(2))and glucose levels in the arterial blood of the rats were detected before tracheotomy and after mechanical ventilation or autonomous respiration for 4 h.Then,the rats were sacrificed and their blood samples were collected to detect the concentrations of IL-1βand IL-18 in the serum and bronchoalveolar lavage fluid(BALF)by enzyme-linked immuno sorbent assay(ELISA).The gross appearance of lung tissue was observed,the pathological morphology of lung tissue was observed by H-E staining under a light microscope,and lung injury scores were evaluated.The wet/dry(W/D)of lung tissue was detected.Western blot was used to detect the levels of NLRP3,GSDMD,apoptosis-related speck protein(ASC),and caspase-1 in rat lung tissues.Results Group C showed nearly normal morphological structure of pulmonary alveoli in rats,while completely destructed pulmonary alveoli were seen in group V,with alveolar breakage and fusion,alveolar septum edema,much inflammatory cell infiltration,and alveolar hemorrhage.Compared with group C,group V and group S pre

关 键 词：西格列汀 机械通气 肺损伤 NOD样受体热蛋白结构域相关蛋白3 消皮素D 

分 类 号：R965[医药卫生—药理学]