摘要
目的评价高迁移率族蛋白B1(HMGB1)在肠缺血再灌注小鼠肺损伤中的作用及其与中性粒细胞胞外诱捕网(NETs)的关系。方法SPF级健康成年雄性C57BL/6小鼠24只,体重20~25 g,采用随机数字表法分为4组(n=6):假手术组(Sham组)、肠缺血再灌注组(I/R组)、IgY对照抗体组(IgY组)和Anti-HMGB1中和抗体组(Anti-HMGB1组)。采用夹闭肠系膜上动脉1 h后恢复灌注的方法制备小鼠肠缺血再灌注损伤模型。Anti-HMGB1组于缺血1 h后肠道血流恢复即刻腹腔注射Anti-HMGB1中和抗体1.0 mg/kg;IgY组于缺血1 h后肠道血流恢复即刻腹腔注射IgY同型对照抗体1.0 mg/kg。于再灌注4 h时处死小鼠,收集小鼠支气管肺泡灌洗液(BALF),测定总蛋白、游离双链DNA(dsDNA)及HMGB1的浓度;取肺组织HE染色后光镜下观察病理学结果,采用Western blot法测定瓜氨酸化组蛋白H3(Cit-H3)表达。结果与Sham组比较,I/R组和IgY组肺组织病理学损伤评分、BALF总蛋白、HMGB1和dsDNA浓度升高,Cit-H3表达上调(P<0.05);与IgY组比较,Anti-HMGB1组肺组织病理学损伤评分、BALF总蛋白、HMGB1和dsDNA浓度降低,肺组织Cit-H3表达下调(P<0.05)。结论HMGB1参与了肠缺血再灌注小鼠肺损伤的过程,与介导NETs形成有关。
Objective To evaluate the role of high-mobility group box 1 protein(HMGB1)in lung injury induced by intestinal ischemia-reperfusion(I/R)in mice and the relationship with neutrophil extracellular traps(NETs).Methods Twenty-four healthy SPF adult male C57BL/6 mice,weighing 20-25 g,were divided into 4 groups(n=6 each)by using a random number table method:sham operation group(Sham group),intestinal I/R group(I/R group),IgY control antibody group(IgY group)and Anti-HMGB1 neutralizing antibody group(Anti-HMGB1 group).Intestinal I/R injury model was established by occlusion of superior mesenteric artery for 1 h followed by reperfusion.Anti-HMGB1 neutralizing antibody 1.0 mg/kg was intraperitoneally injected immediately after restoration of reperfusion in group Anti-HMGB1.An IgY isotype control antibody 1.0 mg/kg was intraperitoneally injected immediately after restoration of reperfusion in group IgY.The animals were sacrificed at the end of reperfusion.Bronchoalveolar lavage fluid(BALF)was collected for determination of total protein and extracellular free double strand DNA(dsDNA)and HMGB1 concentrations.Lung tissues were removed for examination of pathological changes with a light microscope after HE staining and for determination of citrullinated histone H3(Cit-H3)expression by Western blot.The pathological changes of the lung were scored.Results Compared with group Sham,the pathological scores and concentrations of total protein,HMGB1 and dsDNA in BALF were significantly increased,and the expression of Cit-H3 was up-regulated in I/R and IgY groups(P<0.05).Compared with group IgY,the pathological scores and concentrations of total protein,HMGB1 and dsDNA in BALF were significantly decreased,and the expression of Cit-H3 was down-regulated in group Anti-HMGB1(P<0.05).Conclusion HMGB1 is involved in lung injury induced by intestinal I/R in mice,which is related to mediating the formation of NETs.
作者
詹雅清
凌逸虹
赖汉津
丘煜鑫
沈建通
温仕宏
黄文起
Zhan Yaqing;Ling Yihong;Lai Hanjin;Qiu Yuxin;Shen Jiantong;Wen Shihong;Huang Wenqi(Department of Anesthesiology,the First Affiliated Hospital,Sun Yat-Sen University,Guangzhou 510080,China;Department of Pathology,Sun Yat-Sen University Cancer Center,Guangzhou 510060,China;Surgical Intensive Care Unit,the First Affiliated Hospital,Sun Yat-Sen University,Guangzhou 510080,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2020年第10期1263-1265,共3页
Chinese Journal of Anesthesiology
基金
国家自然科学基金(81772116,81801895)
广东省基础与应用基础研究基金(2019A1515010143)。
关键词
高迁移率族蛋白类
胞外诱捕网
肠
再灌注损伤
肺损伤
High mobility group proteins
Extracellular traps
Intestine
Reperfusion injury
Lung injury