摘要
目的:探讨中性粒细胞胞外诱捕网(NETs)在新生大鼠急性肺损伤(ALI)中的作用。方法:取出生7 d的SD大鼠30只,按照随机数字表法分成生理盐水对照组、ALI组及ALI+脱氧核糖核酸酶(Dnase)组,每组10只。ALI组用脂多糖(LPS)以20 mg/kg的剂量腹腔注射,ALI+Dnase组则在注射LPS后即腹腔注射Dnase(5 mg/kg)。给药6 h后,水合氯醛麻醉大鼠,收集支气管肺泡灌洗液(BALF),荧光酶标仪检测BALF中游离DNA(cf-DNA)的含量;右肺组织固定于4%多聚甲醛中,HE染色观察各组大鼠肺组织形态结构;左肺组织制备肺组织匀浆,酶联免疫吸附测定(ELISA)法检测肺组织匀浆中白细胞介素6(IL-6)及肿瘤坏死因子α(TNF-α)的含量;使用免疫荧光法与Western blot检测各组大鼠肺组织中瓜氨酸化组蛋白H3(CitH3)及髓过氧化物酶(MPO)的生成情况。结果:与对照组相比,ALI组与ALI+Dnase组中cf-DNA、CitH3、MPO、IL-6及TNF-α水平均升高(P<0.05),肺组织中炎性细胞浸润严重;与ALI组相比,ALI+Dnase组新生大鼠肺组织中cf-DNA、Cith3、MPO、IL-6及TNF-α水平降低(P<0.05),ALI+Dnase组炎症浸润程度降低。结论:新生大鼠ALI中,NETs水平为反映肺组织损伤的重要指标,NETs可能为治疗新生儿ALI的新靶点。
AIM:To investigate the role of neutrophil extracellular traps(NETs)in neonatal rats with acute lung injury(ALI).METHODS:Thirty 7-day-old SD rats were randomly divided into normal saline control group,ALI group and ALI+deoxyribonuclease(Dnase)group(each n=10).The rats in ALI group were intraperitoneally injected with lipopolysaccharides(LPS)at 20 mg/kg,and the rats in ALI+Dnase group were intraperitoneally injected with Dnase at 5 mg/kg after LPS injection.After 6 h,the rats were anesthetized with chloral hydrate,bronchial alveolar lavage fluid(BALF)was collected,and the content of cell-free DNA(cf-DNA)in BALF was detected by fluorescence microarray.The right lung tissues were fixed in 4% paraformaldehyde,and the morphological structure of the lung tissues were observed by HE staining.The content of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the left lung homogenate was measured by ELISA.Immunofluorescence and Western blot were used to detect the production of citrullinated histone H3(CitH3)and myeloperoxidase(MPO)in the rat lung tissues.RESULTS:Compared with control group,the levels of cf-DNA,CitH3,MPO,IL-6 and TNF-α in lung tissues of neonatal rats in ALI group and ALI+Dnase group were all increased(P<0.05),and severe inflammatory infiltration in the lung tissues was observed.Compared with ALI group,the levels of cf-DNA,CitH3,MPO,IL-6 and TNF-α in ALI+Dnase group were decreased(P<0.05),and the inflammatory infiltration was attenuated.CONCLUSION:In neonatal rats with ALI,the level of NETs is an important indicator of lung tissue injury,and NETs may be a new target for the treatment of neonatal ALI.
作者
徐苏晴
常明
宓兰兰
蔡佳玉
薛正阳
卢红艳
XU Su-qing;CHANG Ming;MI Lan-lan;CAI Jia-yu;XUE Zheng-yang;LU Hong-yan(Department of Pediatrics,Affiliated Hospital of Jiangsu University,Zhenjiang 212001,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2020年第5期913-917,923,共6页
Chinese Journal of Pathophysiology
基金
江苏省妇幼健康重点人才项目(No.FRC201735)
镇江市重点研发计划(社会发展)项目(No.SH2018050)。
