Nrf2/HO-1信号通路在艾司氯胺酮减轻小鼠内毒素性急性肺损伤中的作用:与NLRP3炎性小体介导细胞焦亡的关系  

Role of Nrf2/HO-1 signaling pathway in reduction of endotoxin-induced acute lung injury by esketamine in mice: relationship with NLRP3 inflammasome-mediated pyroptosis

作　　者：马扬 刘静怡[2] 马子健 张继骁 曹雪峰[1] 李艳[1] 王志学[1] Ma Yang;Liu Jingyi;Ma Zijian;Zhang Jixiao;Cao Xuefeng;Li Yan;Wang Zhixue(Department of Anesthesiology,Chengde Medical University Affiliated Hospital,Chengde 067000,China;Department of Cardiology,Chengde Medical University Affiliated Hospital,Chengde 067000,China)

机构地区：[1]承德医学院附属医院麻醉科,承德067000 [2]承德医学院附属医院心脏内科,承德067000

出　　处：《中华麻醉学杂志》2023年第10期1237-1242,共6页Chinese Journal of Anesthesiology

基　　金：承德市基础研究项目(202204A157);河北省科技厅重点研发计划项目(22377746D)。

摘　　要：目的评价NF-E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)信号通路在艾司氯胺酮减轻小鼠内毒素性急性肺损伤中的作用及其与核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎性小体介导细胞焦亡的关系。方法SPF级雄性野生型(WT)及Nrf2敲除型(KO)C57BL/6J小鼠各18只,体质量20~25 g,6~8周龄,采用随机数字表法分别将两种小鼠分为3组(n=6):对照组(WT+C组、KO+C组)、内毒素性急性肺损伤组(WT+ALI组、KO+ALI组)和内毒素性急性肺损伤+艾司氯胺酮组(WT+ALI+E组、KO+ALI+E组)。采用尾静脉注射脂多糖(LPS)15 mg/kg制备小鼠内毒素性急性肺损伤模型。WT+ALI+E组和KO+ALI+E组于注射LPS 30 min后腹腔注射艾司氯胺酮10 mg/kg,6 h后重复给药1次,其余组给予等容量生理盐水。于注射LPS后12 h时麻醉小鼠,心脏穿刺取血样,采用ELISA法测定血清IL-1β、IL-18浓度;取双侧肺脏组织,光镜下观察病理学损伤情况并进行肺损伤评分,微板法测定还原型谷胱甘肽(GSH)含量,采用Western blot方法测定Nrf2、HO-1、NLRP3炎性小体介导的细胞焦亡相关蛋白[NLRP3、凋亡相关斑点样蛋白(ASC)、pro-caspase-1、cleaved-caspase-1、消皮素D(GSDMD)]的表达。结果与相应的C组(WT+C组或KO+C组)比较,WT+ALI组和KO+ALI组肺损伤评分、血清IL-1β和IL-18浓度升高,肺组织GSH含量降低,NLRP3、ASC、pro-caspase-1、cleved-caspase-1和GSDMD表达上调,WT+ALI组肺组织Nrf2和HO-1表达上调(P<0.05);与相应的ALI组(WT+ALI组或KO+ALI组)比较,WT+ALI+E组、KO+ALI+E组肺损伤评分、血清IL-1β和IL-18浓度降低,肺组织GSH含量升高,NLRP3、ASC、pro-caspase-1、cleved-caspase-1和GSDMD表达下调,WT+ALI+E组肺组织Nrf2和HO-1表达上调(P<0.05)。与WT+ALI+E组比较,KO+ALI+E组肺损伤评分、血清IL-1β和IL-18浓度升高,肺组织GSH含量降低,Nrf2和HO-1表达下调,NLRP3、ASC、pro-caspase-1、cleved-caspase-1和GSDMD表达上调(P<0.05)。结论艾司氯胺酮减轻小鼠内毒素性急性肺�Objective To evaluate the role of NF-E2-related factor 2(Nrf2)/heme oxygenase(HO-1)signaling pathway in reduction of endotoxin-induced acute lung injury(ALI)by esketamine and the relationship with NOD-like receptor family pyrin domain containing 3(NLRP3)inflammasome-mediated pyroptosis in mice.Methods SPF male wild-type(WT)and Nrf2 knockout(KO)C57BL/6J mice,aged 6-8 weeks,weighing 20-25 g,were divided into 3 groups(n=6 each)using a random number table method:control group(WT+C group,KO+C group),ALI group(WT+ALI group,KO+ALI group)and ALI+esketamine group(WT+ALI+E group,KO+ALI+E group).ALI model was developed by injection of lipopolysaccharide(LPS)15 mg/kg via the tail vein.Esketamine 10 mg/kg was intraperitoneally injected at 30 min after LPS injection,and 6 h later the medication was repeated for one time in WT+ALI+E and KO+ALI+E groups,while the equal volume of normal saline was given in the other groups.The mice were anesthetized at 12 h after LPS injection,and blood samples were obtained by cardiac puncture for determination of serum interleukin-1beta(IL-1β)and IL-18 concentrations,and bilateral lung tissues were also obtained for examination of the pathological changes of lung tissues(with the light microscope)which were scored and for determination of the content of reduced glutathione(GSH)and expression of Nrf2,HO-1 and NLRP3 inflammasome-mediated pyroptosis-related proteins(NLRP3,apoptosis-associated speck-like protein containing a CARD[ASC],pro-caspase-1,cleaved-caspase-1,gasdermin D[GSDMD])(by Western blot).Results Compared with the corresponding C group(WT+C group or KO+C group),the lung injury score and concentrations of IL-1βand IL-18 were significantly increased,the content of GSH in lung tissues was decreased,and the expression of NLRP3,ASC,pro-caspase-1,cleved-caspase-1 and GSDMD was up-regulated in WT+ALI group and KO+ALI group(P<0.05),and the expression of Nrf2 and HO-1 was significantly up-regulated in WT+ALI group(P<0.05).Compared with the corresponding ALI group(WT+ALI group or KO+ALI group

关 键 词：内毒素血症 急性肺损伤 NF-E2相关因子2 血红素加氧酶-1 NLR家族 热蛋白结构域包含蛋白3 细胞焦亡 艾司氯胺酮 

分 类 号：R614[医药卫生—麻醉学]