双侧颈上交感神经节在小鼠心肌缺血再灌注损伤中的作用:与NLRP3炎症小体的关系  

Role of bilateral superior cervical sympathetic ganglion in myocardial ischemia-reperfusion injury in mice: relationship with NLRP3 inflammasomes

作　　者：赵晓帅 张羽茜 田浩 李璐[1] 邱珍[1] 夏中元[1] Zhao Xiaoshuai;Zhang Yuxi;Tian Hao;Li Lu;Qiu Zhen;Xia Zhongyuan(Department of Anesthesiology,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院麻醉科,武汉430060

出　　处：《中华麻醉学杂志》2023年第5期607-612,共6页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(82172160,81970722);武汉大学自主科研项目(2042021kf0113)。

摘　　要：目的:评价双侧颈上交感神经节在小鼠心肌缺血再灌注损伤中的作用及其与NOD样受体蛋白3(NLRP3)炎症小体的关系。方法:SPF级健康雄性C57BL小鼠32只,8~10周龄,体质量25~30 g,采用随机数字表法分成4组(n=8):假手术组(NS组)、心肌缺血再灌注组(NIR组)、双侧颈上交感神经节切除组(SCGx组)和双侧颈上交感神经节切除+心肌缺血再灌注组(SCGx+IR组)。采用结扎左冠状动脉前降支30 min,再灌注24 h的方法制备小鼠心肌缺血再灌注损伤模型。于再灌注3 d前行双侧颈上交感神经节切除术。于再灌注24 h时采集下腔静脉血样,采用ELISA法测定血清CK-MB活性、cTnI浓度、去甲肾上腺素(NE)浓度和LDH活性,比色法测定SOD活性,DHE法检测心肌ROS水平,TTC法检测心肌梗死面积,HE和WGA染色法观察病理学结果,qPCR法检测IL-1β、IL-6、IL-10、TNF-α和NLRP3的mRNA表达,Western blot法检测酪氨酸羟化酶(TH)、IL-1β、TNF-α、NLRP3、心房钠尿肽(ANP)和脑利钠肽(BNP)的表达。结果:与NS组比较,SCGx组NE浓度降低,TH表达下调,NIR组血清CK-MB活性、cTnI、NE浓度、LDH活性和心肌ROS水平升高,SOD活性降低,IL-1β、TNF-α、NLRP3、ANP和BNP表达上调,IL-1β、IL-6、TNF-α、NLRP3的mRNA表达上调(P<0.05);与SCGx组比较,SCGx+IR组血清CK-MB活性、cTnI、NE浓度、LDH活性和心肌ROS水平升高,SOD活性降低,IL-1β、TNF-α、NLRP3、ANP和BNP表达上调,IL-1β、IL-6、TNF-α和NLRP3的mRNA表达上调(P<0.05);与NIR组比较,SCGx+IR组血清CK-MB活性、cTnI浓度、LDH活性和心肌ROS水平降低,SOD活性升高,IL-1β、TNF-α、NLRP3、ANP和BNP表达下调,IL-1β、IL-6、TNF-α和NLRP3的mRNA表达下调,心肌梗死面积减少(P<0.05)。结论:双侧颈上交感神经节切除减轻小鼠心肌缺血再灌注损伤的机制与降低NLRP3炎症小体活化,抑制炎症反应有关。Objective To evaluate the role of bilateral superior cervical sympathetic ganglia(SCG)in myocardial ischemia-reperfusion(I/R)injury in mice and the relationship with NOD-like receptor protein 3(NLRP3)inflammasomes.MethodsThirty-two healthy SPF male C57BL mice,aged 8-10 weeks,weighing 25-30 g,were divided into 4 groups(n=8 each)by the random number table method:sham operation group(NS group),myocardial I/R group(NIR group),bilateral SCG excision group(SCGx group)and bilateral SCG excision+myocardial I/R group(SCGx+IR group).The myocardial I/R injury model was prepared by ligating the anterior descending branch of the left coronary artery for 30 min followed by 24 h reperfusion in isoflurane-anesthetized mice.Bilateral superior cervical sympathectomy was performed at 3 days before reperfusion.Blood samples were collected from the inferior vena cava at 24 h of reperfusion for examination of pathological changes(by HE and WGA staining)and for measurement of serum creatine kinase isoenzymes(CK-MB)activity,cardiac troponin I(cTnI)concentration,norepinephrine(NE)concentration and lactic dehydrogenase(LDH)activity(by enzyme-linked immunosorbent assay),superoxide dismutase(SOD)activity(by colorimetric method),myocardial reactive oxygen species(ROS)level(by DHE method),myocardial infarct size(by TTC method),and expression of interleukin-1beta(IL-1β),IL-6,IL-10,tumor necrosis factor-alpha(TNF-α),NLRP3 mRNA(by quantitativepolymerase chain reaction),and expression of tyrosine hydroxylase(TH),IL-1β,TNF-α,NLRP3,atrial natriuretic peptide(ANP)and brain natriuretic peptide(BNP)(by Western blot).ResultsCompared with NS group,the NE concentration was significantly decreased,and TH expression was down-regulated in SCGx group,and the serum CK-MB activity,concentrations of cTnI and NE,LDH activity and myocardial ROS level were significantly increased,SOD activity was decreased,the expression of IL-1β,TNF-α,NLRP3,ANP and BNP was up-regulated,and the expression of IL-1β,IL-6,TNF-αand NLPR3 mRNA was up-regulated in NIR group(P<0.

关 键 词：颈上神经节 交感神经切除术 心肌再灌注损伤 NLR家族 热蛋白结构域包含蛋白3 

分 类 号：R614[医药卫生—麻醉学]