摘要
目的:探讨氢气对脂多糖(lipopolysaccharide, LPS)诱导的脓毒症小鼠急性肺损伤(acute lung injury, ALI)的保护作用及对NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3, NLRP3)/组织因子(tissue factor, TF)信号通路表达的影响。方法:雄性C57BL/6J小鼠64只,分2批进行实验。每批小鼠采用随机数字表法分为4组:空白对照组(C组)、LPS组(L组)、LPS+氢气组(LH组)和氢气组(H组),每组8只。C组和H组腹腔注射10 ml/kg生理盐水,L组和LH组腹腔注射LPS 10 mg/kg建立脓毒症ALI模型。C组和L组常规饲养12 h,LH组和H组吸入4%氢气治疗12 h。造模12 h后,第1批小鼠用于肺组织病理检查、肺功能检测和散斑多普勒血流成像,第2批小鼠用于支气管肺泡灌洗和Western blot检测。H-E染色观察肺组织病理学特征并进行肺组织损伤评分,全身体积描记系统检测小鼠肺功能指标[每分通气量(minute ventilation volume, MV)、呼气中期流速(expiratory flow 50, EF50)、最大呼气流速(peak expiratory flow, PEF)、气道狭窄指数(enhanced pause, Penh)],散斑多普勒血流成像检测小鼠右肺和左后足血流量,Western blot检测小鼠肺组织中NLRP3炎症小体和TF的含量,ELISA法检测小鼠支气管肺泡灌洗液(bronchoalveolar lavage fluid, BALF)中TNF-α和IL-6的浓度。结果:H-E染色显示C组和H组小鼠肺组织结构正常;L组小鼠肺组织结构紊乱,大量中性粒细胞浸润;LH组小鼠肺组织中性粒细胞浸润减少,水肿减轻。与C组比较,L组和LH组肺组织损伤评分明显升高( P<0.05);MV明显下降,EF50和PEF明显降低,Penh明显升高(均 P<0.05);右肺和左后足血流量明显减少( P<0.05);肺组织NLRP3和TF的相对含量明显升高( P<0.05);BALF中TNF-α和IL-6浓度明显升高( P<0.05)。与C组比较,H组各指标差异均无统计学意义( P>0.05)。与L组比较,LH组小鼠肺组织损伤评分明显降低( P<0.05);MV明显升高,PEF明显升高,Penh明�
Objective To investigate the protective effect of hydrogen on lipopolysaccharide(LPS)‐induced acute lung injury(ALI)in septic mice and the effect on the expression of the NOD‐like receptor thermal protein domain associated protein 3(NLRP3)/tis‐sue factor(TF)signaling pathway.Methods A total of 64 male C57BL/6J mice were tested in two batches.According to the random number table method,mice in each batch were divided into four groups(n=8):a control group(group C),a LPS group(group L),a LPS+hydrogen group(group LH)and a hydrogen group(group H).Mice in groups C and H were intraperitoneally injected with normal saline at 10 ml/kg.Meanwhile,those in groups L and LH were intraperitoneally injected with 10 mg/kg LPS to establish an ALI model in sep‐tic mice.Groups C and L were conventionally housed for 12 h,while groups LH and H were inhaled with 4%hydrogen for 12 h.Then,12 h after modeling,mice in batch 1 were used for lung histopathology,lung function tests and scattered Doppler flow imaging,while those in batch 2 were used for bronchoalveolar lavage and Western blot analysis.hematoxylin‐eosin(H‐E)staining was performed to ob‐serve lung histopathology and evaluate lung injury score.Whole‐body plethysmography system was used to detect lung function indica‐tors in mice:minute ventilation volume(MV),expiratory flow 50(EF50),peak expiratory flow(PEF),and enhanced pause(Penh).Scat‐tered Doppler flow imaging was conducted to detect blood flow in the right lung and the left hind foot of mice.The levels of NLRP3 in‐flammatory vesicles and TF in the lungs were measured by Western blot.The levels of inflammatory factors tumor necrosis factor‐α(TNF‐α)and interleukin(IL)‐6 in bronchoalveolar lavage fluid(BALF)of mice were detected by enzyme‐linked immunosorbent assay(ELISA).Results According to H‐E staining,the lung tissue structure of mice in group C and group H was normal.The lung tissue structure of mice in group L was disorganized,with a large number of neutrophils infiltrating.The lung tissu
作者
昂扬
李茜
施敏
孙涛
龙云
段满林
Ang Yang;Li Qian;Shi Min;Sun Tao;Long Yun;Duan Manlin(Department of Anesthesiology,Jinling Hospital,Affiliated Hospital of Medical School,Nanjing University,Nanjing 210002,China;Department of Anesthesiology,the Affiliated Jiangning Hospital of Nanjing Medical University,Nanjing 211100,China)
出处
《国际麻醉学与复苏杂志》
CAS
2023年第11期1126-1132,共7页
International Journal of Anesthesiology and Resuscitation
基金
南京市医学科技发展项目(YKK21226)
南京医科大学附属江宁医院免疫细胞转化研究中心开放课题(JNYYZXKY202216)
江苏大学医教协同创新基金一般项目(JDYY2023094)
2021年江宁区科技惠民计划项目(20212021NJN‐QKJHMJHXM0133)。
关键词
脓毒症
急性肺损伤
氢气
NOD样受体热蛋白结构域相关蛋白3
组织因子
Sepsis
Acute lung injury
Hydrogen
NOD‐like receptor thermal protein structural domain associated protein 3
Tissue factor
