甲烷对脊髓缺血再灌注大鼠炎症反应时P2X7R/NLRP3信号通路的影响  

Effect of methane on P2X7R/NLRP3 signaling pathway during inflammatory responses in a rat model of spinal cord ischemia-reperfusion

作　　者：曾昭恺 薛海燕 王丽萍 Zeng Zhaokai;Xue Haiyan;Wang Liping(Department of Anesthesia and Perioperative Medicine,Fuzhou General Clinical Medical College of Fujian Medical University 900th Hospital of Joint Logistics Support Force Dongfang Hospital Affiliated to Xiamen University Fuzhou General Teaching Hospital of Fujian University of Traditional Chinese Medicine Fuzhou General Teaching Hospital of Bengbu Medical College,Fuzhou 350025,China)

机构地区：[1]福建医科大学福总临床医学院、联勤保障部队第九〇〇医院、厦门大学附属东方医院、福建中医药大学福总教学医院、蚌埠医学院福总教学医院麻醉与围术期医学科,福州350025

出　　处：《中华麻醉学杂志》2020年第2期225-228,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金青年基金(31700740);福建省科技创新联合资金项目(2017Y9126);福建医科大学大学生创新创业训练计划资助项目(C18030)。

摘　　要：目的评价甲烷对脊髓缺血再灌注大鼠炎症反应时嘌呤能受体P2X配体门控离子通道7(P2X7R)/NOD样受体蛋白3(NLRP3)信号通路的影响。方法清洁级健康成年雄性SD大鼠54只,3月龄,体重350~380 g,采用随机数字表法分为3组(n=18):假手术组(S组)、脊髓缺血再灌注组(I/R组)和甲烷组(M组)。S组经右髂总动脉逆行置入Fogarty球囊套管至胸主动脉但不阻断缺血;I/R组采用阻断胸主动脉联合体循环低血压的方法诱导脊髓缺血9 min,然后恢复灌注,建立脊髓缺血再灌注损伤模型;M组于再灌注即刻腹腔注射10 ml/kg甲烷饱和生理盐水。于再灌注12、24和48 h时测定后肢运动-感觉障碍指数(MSDI);于再灌注48 h时处死大鼠取L3-5脊髓组织,采用尼氏染色和免疫荧光染色确定脊髓前角和后角正常神经元数量、活化小胶质细胞数量及其表达P2X7R、NLRP3、凋亡相关斑点样蛋白(ASC)、计caspase-1、IL-1β和IL-18的水平。结果与S组比较,I/R组再灌注12、24和48 h时后肢MSDI升高,再灌注48 h时脊髓前后角正常神经元计数减少,活化小胶质细胞计数增加,P2X7R、NLRP3、ASC、caspase-1、IL-1β和IL-18表达上调(P<0.01);与I/R组比较,M组再灌注各时点后肢MSDI降低,再灌注48 h时脊髓前后角正常神经元计数增加,活化小胶质细胞计数减少,P2X7R、NLRP3、ASC、caspase-1、IL-1β和IL-18表达下调(P<0.05)。结论甲烷减轻脊髓缺血再灌注大鼠炎症反应的机制与抑制P2X7R/NLRP3信号通路有关。Objective To evaluate the effect of methane on purinergic ion channel type 7 receptor(P2X7R)/nod-like receptor protein 3(NLRP3)signaling pathway during inflammatory responses in a rat model of spinal cord ischemia-reperfusion(I/R).Methods Fifty-four clean-grade healthy male Sprague-Dawley rats,aged 3 months,weighing 350-380 g,were divided into 3 groups(n=18 each)using a random number table method:sham operation group(group S),spinal cord I/R group(group I/R)and methane group(group M).Rats underwent sham operation in group S.Spinal cord ischemia was induced by occlusion of the thoracic aorta combined with controlled hypotension for 9 min,followed by reperfusion in anesthetized animals in group I/R.Methane-rich saline 10 mg/kg was intraperitoneally administered immediately after onset of reperfusion in group M.Motor sensory deficit index(MSDI)in hind limbs was measured at 12,24 and 48 h of reperfusion.The L3-5 segment of spinal cord was removed at 48 h of reperfusion for determination of the number of normal neurons(by Nissl staining),the number of activated microglia and expression of P2X7R,NLRP3,apoptosis-associated speck-like protein containing CARD(ASC),cysteine-requiring aspartate protease(caspase-1),interleukin-1beta(IL-1β)and IL-18(by immunofluorescence staining)in anterior and posterior horns of spinal cord.Results Compared with group S,the MSDI was significantly increased at 12,24 and 48 h of reperfusion,and the number of normal neurons in anterior and posterior horns of spinal cord was decreased,the number of activated microglia was increased,and the expression of P2X7R,NLRP3,ASC,caspase-1,IL-1βand IL-18 was up-regulated at 48 h of reperfusion in group I/R(P<0.01).Compared with group I/R,the MSDI was significantly decreased at each time point of reperfusion,and the number of normal neurons in anterior and posterior horns of spinal cord was increased,the number of activated microglia was decreased,and the expression of P2X7R,NLRP3,ASC,caspase-1,IL-1βand IL-18 was down-regulated at 48 h of reperfusion in

关 键 词：甲烷 受体 嘌呤能P2 NLR家族 热蛋白结构域包含蛋白3 脊髓 再灌注损伤 炎症 

分 类 号：R654.3[医药卫生—外科学]