摘要
目的 观察冷缺血期一氧化碳(CO)和氢气(H_(2))联合膨肺对大鼠移植肺脏炎症反应的影响及机制。方法 选择成年SPF级雄性SD大鼠80只,40只供体和40只受体,8~10周龄,体重250~280 g。采用随机数字表法将大鼠分为四组:O_(2)膨肺组(O_(2)组)、CO膨肺组(CO组)、H_(2)膨肺组(H_(2)组)和CO和H_(2)联合膨肺组(CH组),每组10对(每对1只供体和1只受体)。供体大鼠切取左肺后,于冷缺血期进行气体膨肺,O_(2)组采用40%O_(2)+60%N_(2)膨肺,CO组采用0.05%CO+40%O_(2)+59.95%N_(2)膨肺,H_(2)组采用3%H_(2)+40%O_(2)+57%N_(2)膨肺,CH组采用0.05%CO+3%H_(2)+40%O_(2)+56.95%N_(2)膨肺,四组体积均为5 ml/kg,每30分钟使用气密针置换气体1次,180 min后行肺移植。受体大鼠于移植前即刻、再灌注后3、60、120、180 min进行血气分析,记录PaO_(2)/FiO_(2)、pH和碱剩余(BE)。再灌注后180 min处死受体大鼠,测定移植肺组织湿/干重比(W/D),采用ELISA法检测移植肺组织白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)浓度及髓过氧化物酶(MPO)含量。检测移植肺组织支气管肺泡灌洗液(BALF)中性粒细胞百分比,采用高效液相色谱法检测BALF中大聚积体(LA)、小聚积体(SA)和LA/SA。采用Western blot法检测移植肺组织表面活性蛋白A(SP-A)和核转录因子-κB(NF-κB)蛋白含量。采用HE染色法观察移植肺组织病理形态并进行肺组织损伤评分(LIS)。结果 与O_(2)组比较,CO组、H_(2)组和CH组受体大鼠再灌注后60、120、180 min PaO_(2)/FiO_(2)、pH、BE均明显升高(P<0.05),移植肺组织W/D、IL-6、IL-1β、TNF-α浓度、MPO含量、中性粒细胞百分比、SA、NF-κB蛋白含量和LIS均明显降低(P<0.05),IL-10浓度、LA、LA/SA和SP-A蛋白含量明显升高(P<0.05)。与CO组比较,H_(2)组和CH组受体大鼠再灌注后120、180 min PaO_(2)/FiO_(2)、pH、BE均明显升高(P<0.05),CH组移植肺组织W/D、IL-6、IL-1β、TNF-α浓度、
Objective To observe the effect and mechanism of lung inflation with both carbon monoxide(CO) and hydrogen(H_(2)) during the cold ischemia phase(CIP) on the inflammatory response in a model of rat lung transplantation.Methods Eighty SPF SD rats,aged 8-10 weeks,weighing 250-280 g,including donors and recipients,were randomly divided into four groups:O_(2) inflation group(group O_(2)),CO inflation group(group CO),H_(2) inflation group(group H_(2)),and CO combined H_(2) inflation group(group CH),10 pairs of rats in each group(one donor and one recipient in each pair).After the donor lungs harvested,they were inflated during the CIP with 40%O_(2) + 60%N_(2),0.05%CO + 40%O_(2) + 59.95%N_(2),3%H_(2) + 40%O_(2) + 57%N_(2),or 0.05%CO + 3%H_(2) + 40%O_(2) + 56.95%N_(2),respectively(5 ml/kg).The mixed gas was replaced every 30 minutes,and lung transplantation was performed 180 minutes later.The blood gas analysis such as PaO_(2)/FiO_(2),pH,and base excess(BE) was performed before transplantation and 3,60,120,and 180 minutes after reperfusion in the recipient.The lung wet/dry ratio(W/D),the myeloperoxidase(MPO),interleukin-6(IL-6),interleukin-10(IL-10),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α) in lung tissues detected by ELISA,the percentage of neutrophil in bronchoalveolar lavage fluid(BALF),the large surfactant aggregate(LA),small surfactant aggregate(SA),and LA/SA were detected by high-performance liquid chromatography,the pulmonary surfactant associated protein(SP)-A and nuclear factor-κB(NF-κB) protein expression were detected by western blot,and the lung injury score(LIS) were measured by HE 180 minutes after reperfusion.ResultsCompared with group O_(2),the PaO_(2)/FiO_(2),pH,and BE 60,120,and 180 minutes after reperfusion in recipients were significantly increased(P < 0.05),transplanted lung tissue W/D,the concentration of IL-6,IL-1β,TNF-α,and MPO,neutrophil percentage,SA,NF-κB protein content,and LIS were significantly decreased(P < 0.05),while the concentration of IL-10,LA,LA/SA,and SP-A protein
作者
刘思同
郭力甲
孟超
康继宇
蔡振华
LIU Sitong;GUO Lijia;MENG Chao;KANG Jiyu;CAI Zhenhua(Department of Anesthesiology,the First Affiliated Hospital of Harbin Medical University,Harbin 150001,China)
出处
《临床麻醉学杂志》
CAS
CSCD
北大核心
2024年第4期401-407,共7页
Journal of Clinical Anesthesiology
关键词
一氧化碳
氢气
联合膨肺
冷缺血期
肺移植
肺损伤
大鼠
Carbon monoxide
Hydrogen
Combination inflation
Cold ischemia
Lung transplantation
Lung injury
Rats
