内质网IRE1α/XBP1信号通路在小鼠内毒素性急性肺损伤中的作用:与NLRP3炎症小体的关系  

Role of IRE1α/XBP1 signaling pathway in endoplasmic reticulum in endotoxin-induced acute lung injury in mice: relationship with NLRP3 inflammasomes

作　　者：毕夏 孙莉莉 王宜博 张晓 孙立新[1] 马福国[1] 韩伟[3] Bi Xia;Sun Lili;Wang Yibo;Zhang Xiao;Sun Lixin;Ma Fuguo;Han Wei(Department of Anesthesiology,Qingdao Municipal Hospital,Qingdao 266071,China;Department of Nephrology and Immunology,Qingdao Women and Children′s Hospital,Qingdao 266034,China;Department of Respiratory and Critical Care Medicine of Qingdao Municipal Hospital,Qingdao 266071,China)

机构地区：[1]青岛市市立医院麻醉科,青岛266071 [2]青岛市妇女儿童医院肾脏免疫科,青岛266034 [3]青岛市市立医院呼吸与危重症医学科,青岛266071

出　　处：《中华麻醉学杂志》2023年第10期1243-1247,共5页Chinese Journal of Anesthesiology

摘　　要：目的评价内质网肌醇需要酶1α/X盒结合蛋白1(IRE1α/XBP1)信号通路在小鼠内毒素性急性肺损伤(ALI)中的作用及其与NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体的关系。方法SPF级健康雄性C57BL/6小鼠36只,6~8周龄,体质量25~30 g,采用随机数字表法分为3组(n=12):对照组(C组)、内毒素性ALI组(ALI组)和内毒素性ALI+STF-083010组(ST组)。ALI组和ST组雾化吸入LPS 3 mg/ml 30 min制备小鼠内毒素性ALI模型,C组雾化吸入等量生理盐水,ST组于雾化吸入LPS前1 h时腹腔注射IRE1α/XBP1信号通路阻断剂STF-08301050 mg/kg,其余2组腹腔注射等容量生理盐水。LPS或生理盐水雾化吸入后24 h时处死小鼠,收集支气管肺泡灌洗液(BALF)并取肺组织标本,肺组织HE染色后光镜观察病理学结果,行肺损伤评分并计算肺湿重/干重(W/D)比值;ELISA法测定BALF上清液IL-1β和IL-18浓度;Western blot法检测肺组织p-IRE1α、XBP1s、NLRP3、ASC和caspase-1的表达。结果与C组比较,ALI组和ST组肺W/D比值、肺损伤评分、BALF IL-1β和IL-18浓度升高,肺组织p-IRE1α、XBP1s、NLRP3、ASC和caspase-1表达上调(P<0.001);与ALI组比较,ST组肺W/D比值、肺损伤评分、BALF IL-1β和IL-18浓度降低、肺组织p-IRE1α、XBP1s、NLRP3、ASC和caspase-1表达下调(P<0.001)。结论IRE1α/XBP1信号通路参与了小鼠内毒素性ALI的过程,其机制与活化NLRP3炎症小体有关。Objective To evaluate the role of inositol-requiring kinase 1α-X-box binding protein 1(IRE1α/XBP1)signaling pathway in endoplasmic reticulumin in endotoxin-induced acute lung injury(ALI)in mice and the relationship with NOD-like receptor pyrin domain containing 3(NLRP3)inflammasomes.Methods Thirty-six SPF-grade healthy male C57BL/6 mice,aged 6-8 weeks,weighing 25-30 g,were divided into 3 groups(n=12 each)according to the random number table method:control group(group C),endotoxin-induced ALI group(group ALI)and endotoxin-induced ALI+STF-083010 group(group ST).The ALI model was established by inhaling nebulized lipopolysaccharide(LPS)3 mg/ml for 30 min in ALI and ST groups,while the equal volume of nebulized normal saline was given in group C.IRE1α/XBP1 signaling pathway inhibitor STF-08301050 mg/kg was intraperitoneally injected at 1 h before inhaling LPS in group ST,while the remaining two groups received the equal volume of normal saline intraperitoneally.Mice were sacrificed at 24 h after inhaling nebulized LPS or normal saline,bronchoalveolar lavage fluid(BALF)were collected and lung tissues were removed for microscopic examination of the pathological changes(by HE staining)which were scored and for determination of wet/dry lung weight ratio(W/D ratio),concentrations of interleukin-1beta(IL-lβ)and IL-18 in BALF(by enzyme-linked immunosorbent assay)and expression of phosphorylated IRE1α(p-IRE1α),XBP1s,NLRP3,ASC and caspase-1 in lung tissues(by Western blot).Results Compared with group C,the W/D ratio,lung injury score and concentrations of IL-1βand IL-18 in BALF were significantly increased,and the expression of p-IRE1α,XBP1s,NLRP3,ASC and caspase-1 in lung tissues was up-regulated in group ALI and group ST(P<0.001).Compared with group ALI,the W/D ratio,lung injury score and concentrations of IL-1βand IL-18 in BALF were significantly decreased,and the expression of p-IRE1α,XBP1s,NLRP3,ASC and caspase-1 protein in lung tissues was down-regulated in group ST(P<0.001).Conclusions IRE1α/XBP1 signaling p

关 键 词：内质网 X盒结合蛋白1 内毒素类 急性肺损伤 NLR家族 热蛋白结构域包含蛋白3 

分 类 号：R563.8[医药卫生—呼吸系统]