外周血中5-羟色胺通过诱导中性粒细胞胞外诱捕网生成加重脓毒症小鼠肺损伤  

Peripheral 5-hydroxytryptophan aggravates lung injury in septic mice by inducing the formation of neutrophils extracellular trap

作　　者：黄雨朦 姬倩 朱艳艳 刘大东[2] 付胜巧 王旭[3] 邰宁正[4] Huang Yumeng;Ji Qian;Zhu Yanyan;Liu Dadong;Fu Shengqiao;Wang Xu;Tai Ningzheng(Jiangsu University,Zhenjiang 212001,Jiangsu,China;Department of Intensive Care Unit,Affiliated Hospital of Jiangsu University,Zhenjiang 212001,Jiangsu,China;Department of Oncology,Affiliated Hospital of Jiangsu University,Zhenjiang 212001,Jiangsu,China;Department of Burn Plastic Surgery,Affiliated Hospital of Jiangsu University,Zhenjiang 212001,Jiangsu,China)

机构地区：[1]江苏大学,江苏镇江212001 [2]江苏大学附属医院重症医学科,江苏镇江212001 [3]江苏大学附属医院放疗科,江苏镇江212001 [4]江苏大学附属医院烧伤整形科,江苏镇江212001

出　　处：《中华危重病急救医学》2021年第12期1423-1427,共5页Chinese Critical Care Medicine

基　　金：国家自然科学基金(81701945)。

摘　　要：目的观察外周5-羟色胺(5-HT)诱导中性粒细胞胞外诱捕网(NET)的生成对脓毒症小鼠肺损伤的影响。方法选择6~8周龄野生型(WT型)和Tph1基因敲除(Tph1^(KO))C57小鼠,按随机数字表法分为WT小鼠假手术组、WT小鼠脓毒症组和Tph1^(KO)小鼠假手术组、Tph1^(KO)小鼠脓毒症组。假手术组小鼠接受假手术处理(仅打开腹腔翻动盲肠但不结扎和穿刺,然后关腹);脓毒症组小鼠接受盲肠结扎穿孔术(CLP)建立脓毒症模型。术后12 h处死小鼠,采用酶联免疫吸附试验(ELISA)检测小鼠支气管肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)含量;同时收集小鼠肺组织标本,光镜下观察肺组织病理学改变,采用免疫荧光显微镜观察小鼠肺组织中NET生成情况。结果病理学结果提示:假手术组小鼠肺组织结构完好,未见渗出;而脓毒症组小鼠肺泡结构破坏,肺泡腔内可见明显渗出,肺泡壁增厚并伴有大量炎性细胞浸润,且WT小鼠脓毒症组肺损伤程度较Tph1^(KO)小鼠脓毒症组严重。ELISA结果显示:不同品系假手术组小鼠BALF中TNF-α和IL-6含量比较差异无统计学意义;而脓毒症组小鼠BALF中TNF-α和IL-6含量均较假手术组明显升高〔WT小鼠:TNF-α(μg/L)为158.20±28.46比14.00±3.28,IL-6(μg/L)为304.98±21.78比57.70±12.30;Tph1^(KO)小鼠:TNF-α(μg/L)为85.88±20.13比14.95±1.53,IL-6(μg/L)为169.50±45.61比55.05±12.68,均P<0.01〕,且WT小鼠脓毒症组上述指标水平均明显高于Tph1^(KO)小鼠脓毒症组〔TNF-α(μg/L):158.20±28.46比85.88±20.13,IL-6(μg/L):304.98±21.78比169.50±45.61,均P<0.01〕。免疫荧光染色显示:假手术组小鼠肺脏中均检测到极少量的NET生成;而脓毒症组小鼠肺组织中均检测到大量的NET生成,明显高于假手术组〔WT小鼠:(34.75±7.27)%比(1.75±0.96)%,Tph1^(KO)小鼠:(14.25±5.74)%比(2.50±1.29)%,均P<0.01〕,且WT小鼠脓毒症组小鼠肺脏组织中NET生成量较Tph1^(KO)小鼠脓毒症组明�Objective To observe the effect of peripheral 5-hydroxytryptophan(5-HT)-induced neutrophil extracellular trap(NET)on lung injury in septic mice.Methods Wild-type(WT type)and Tph1 knockout(KO)C57 mice(6-8 weeks)were selected and divided into WT mice sham group,WT mice sepsis group,Tph1^(KO) mice sham group and Tph1^(KO) mice sepsis group according to the random number table method.Mice in the sham group received sham surgery(only open the abdominal cavity to flip the cecum without ligation and puncture,and then close the abdominal cavity);the mice in the sepsis group received cecal ligation and puncture(CLP)to establish sepsis model.The mice were sacrificed 12 hours after the operation,and the levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in bronchialalveolar lavage fluid(BALF)were detected by enzyme linked immunoadsordent assay(ELISA);at the same time,the lung tissues were collected,and the pathological changes of lung tissues were observed under light microscope,and the production of NET in lung tissues was observed by immunofluorescence microscope.Results The pathological results suggested that the lung tissue structure in sham groups was intact without exudation,while the alveolar structures of mice in the sepsis groups were damaged,with obvious exudation in the alveolar cavity and thickened alveolar walls accompanied by a large number of inflammatory cell infiltration,and the degree of lung injury in the sepsis group of WT mice was more severe than that of the sepsis group of Tph1^(KO) mice.ELISA results showed that there was no statistically significant difference in the contents of TNF-αand IL-6 in mice BALF from different strains of the sham group;while the contents of TNF-αand IL-6 in BALF of septic mice group were significantly higher than those in sham group[WT mice:TNF-α(μg/L)was 158.20±28.46 vs.14.00±3.28,IL-6(μg/L)was 304.98±21.78 vs.57.70±12.30;Tph1^(KO) mice:TNF-α(μg/L)was 85.88±20.13 vs.14.95±1.53,IL-6(μg/L)was 169.50±45.61 vs.55.05±12.68,all P<0.01],and the abov

关 键 词：脓毒症 5-羟色胺 中性粒细胞胞外诱捕网 肺损伤 

分 类 号：R459.7[医药卫生—急诊医学]