冷缺血期氢气膨肺对大鼠移植肺NLRP3炎症小体活性的影响  

Effect of lung inflation with hydrogen during cold ischemia phase on NLRP3 inflammasome activity in transplanted lungs of rats

作　　者：郑盼盼 全莉妮 邢恩桐 郑彬 周华成[1] Zheng Panpan;Quan Lini;Xing Entong;Zheng Bin;Zhou Huacheng(Department of Anesthesiology,the Fourth Affiliated Hospital of Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学附属第四医院麻醉科,150001

出　　处：《中华麻醉学杂志》2020年第12期1499-1502,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81570088)。

摘　　要：目的评价冷缺血期氢气膨肺对大鼠移植肺NOD样受体蛋白3(NLRP3)炎症小体活性的影响。方法清洁级健康雄性Wistar大鼠56只,8~10周龄,体重250~300 g,其中24只大鼠作为供体。取32只大鼠为受体,采用随机数字表法分为4组(n=8):假手术组(Sham组)行侧开胸手术,但不进行肺移植术;肺移植组(LT组)供肺冷缺血期不膨肺,进行肺移植术;氧气组(O组)和氢气组(H组)供肺冷缺血期分别以5 ml/kg的40%氧气-60%氮气和3%氢气-40%氧气-57%氮气膨肺,每20 min置换1次,持续120 min后行肺移植术。于移植前、再灌注3、60和120 min(T0~T3)时行动脉血气分析,计算氧合指数(OI),记录pH值和碱剩余(BE)。T3时取血结束后,放血法处死大鼠,光镜下观察移植肺组织病理学结果,并行肺组织损伤评分(LIS),Western blot法测定移植肺NLRP3、凋亡相关微粒蛋白(ASC)和含半胱氨酸的天冬氨酸蛋白水解酶(caspase)-1 p20表达。结果与Sham组比较,LT组T1~3时OI、T2,3时pH值和BE降低,LIS升高,NLRP3、ASC和caspase-1 p20表达上调(P<0.05);与LT组比较,O组和H组T2,3时OI和BE升高,O组T3时和H组T2,3时的pH值升高,LIS降低,H组NLRP3、ASC和caspase-1 p20表达下调(P<0.05),O组NLRP3、ASC和caspase-1 p20表达差异无统计学意义(P>0.05);与O组比较,H组T2,3时BE升高,LIS降低,NLRP3、ASC和caspase-1 p20表达下调(P<0.05),OI和pH值差异无统计学意义(P>0.05)。结论冷缺血期氢气膨肺减轻大鼠移植肺损伤的机制可能与抑制NLRP3炎症小体活性有关。Objective To evaluate the effect of lung inflation with hydrogen during cold ischemia phase on NOD-like receptor protein 3(NLRP3)inflammasome activity in the transplanted lungs of rats.Methods Fifty-six clean-grade healthy male Wistar rats,aged 8-10 weeks,weighing 250-300 g,were selected,of which 24 rats served as donors.Thirty-two rats served as receptors and were divided into 4 groups(n=8 each)using a random number table method:sham operation group(group Sham),lung transplant group(group LT),oxygen group(group O),and hydrogen group(group H).Sham group underwent a left thoracotomy but no transplantation.The left donor lung was deflated during the cold storage phase,and the orthotopic left lung transplantation was perofrmed in group LT.In group O and group H,the left donor lung was inflated with the mixture of 40%O2-60%N2 and 3%H2-40%O2-57%N2 at 5 ml/kg,respectively,during the cold storage phase,the gas was replaced every 20 min,lasting for 120 min,and then the orthotopic left lung transplantation was performed.The arterial blood gas analysis was performed before transplantation(T0)and at 3,60 and 120 min of reperfusion(T1-3)for determination of the oxygenation index(OI),pH value,and base excess(BE).The rats were sacrificed after blood collection at T3,and the transplanted lung tissues were obtained for examination of pathological changes which were scored(lung injury score[LIS])and for determination of the expression of NLRP3,apoptosis-associated speck-like protein(ASC),and cysteinyl aspartate specific proteinase(caspase)-1 p20(by Western blot).Results Compared with Sham group,OI at T1-3 and pH value and BE at T2,3 were significantly decreased,LIS was increased,and the expression of NLRP3,ASC,and caspase-1 p20 was up-regulated in group LT(P<0.05).Compared with group LT,OI and BE values at T2,3 were significantly increased in O and H groups,pH value at T3 in group O and at T2,3 in group H were significantly increased,LIS was decreased in O and H groups,the expression of NLRP3,ASC,and caspase-1 p20 was significant

关 键 词：氢 肺移植 NLR家族 热蛋白结构域包含蛋白3 

分 类 号：R655.3[医药卫生—外科学]