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张毅,程晨,苏景超,张新芳,刘心月,项水英,王彩云,李尹,林先刚,刘自兵.急性肺损伤大鼠模型制备及不同时段肺损伤比较[J].中国实验动物学报,2021,29(1):27~34.
急性肺损伤大鼠模型制备及不同时段肺损伤比较
Preparation of rat model of acute lung injury and comparison of injury at different periods
投稿时间:2020-09-07  
DOI:10. 3969 / j.issn.1005-4847. 2021. 01. 004
中文关键词:  急性肺损伤  模型  气管滴注法  脂多糖
英文关键词:acute lung injury  model  tracheal instillation  lipopolysaccharide
基金项目:
作者单位E-mail
张毅 安徽中医药大学研究生院,合肥 230038 644689045@ qq.com 
程晨 安徽中医药大学研究生院,合肥 230038  
苏景超 安徽中医药大学中西医结合学院,合肥 230038  
张新芳 安徽中医药大学中西医结合学院,合肥 230038  
刘心月 安徽中医药大学针灸推拿学院针灸经络研究所,合肥 230038  
项水英 3.安徽中医药大学针灸推拿学院针灸经络研究所,合肥 230038
4.针灸基础与技术安徽省重点实验室,合肥 230038 
 
王彩云 安徽中医药大学研究生院,合肥 230038  
李尹 安徽中医药大学研究生院,合肥 230038  
林先刚 3.安徽中医药大学针灸推拿学院针灸经络研究所,合肥 230038
4.针灸基础与技术安徽省重点实验室,合肥 230038 
 
刘自兵 2.安徽中医药大学中西医结合学院,合肥 230038
3.安徽中医药大学针灸推拿学院针灸经络研究所,合肥 230038
4.针灸基础与技术安徽省重点实验室,合肥 230038 
zibingliu@ 163.com 
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中文摘要:
       目的 通过气管滴注脂多糖(lipopolysaccharide,LPS)构建急性肺损伤大鼠模型,观察不同时段肺损伤变化规律。 方法 32 只健康 SD 大鼠随机分为正常组、模型组,正常组 8 只大鼠,模型组 24 只大鼠。 模型组依据 LPS 滴注时长不同分为三个亚组:3 h 组、6 h 组和 12 h 组,每组 8 只。 以 LPS(2 mg / kg)暴露式气管滴注法复制 ALI 大鼠模型。 通过大鼠一般状况、肺大体观察、肺功能检测、计算肺湿干重比值、支气管肺泡灌洗液(BALF)中白细胞 介素-1β(IL-1β)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)检测和肺组织中丙二醛(MDA)、超氧化物歧化酶(SOD)检测、苏木素-伊红染色(HE 染色)组织形态学观察,评估不同时段急性肺损伤伤情变化情况。 结果 造模后模型组大鼠存活率为 100%。 大鼠一般状况显示,3 h 组与正常组相比,精神处于淡漠状态、摄食减少、活动明显减少、鼻腔处粘液分泌物增多、大鼠的呼吸频率加快、可闻及哮鸣音。 肺大体观察显示,3 h 组左右肺门处可见肺组织肝样变,左右肺叶上散布出血点,出血部位颜色鲜红。 肺功能显示 3 h 组大鼠第 0. 1 秒用力呼气量( FEV 0. 1)、第 0. 3 秒用力呼气量(FEV 0. 3)以及各占用力肺活量(FVC)之比(FEV0. 1 / FVC;FEV0. 3 / FVC)均显著下降 (P< 0. 05,P< 0. 01);肺湿干重比值明显升高(P< 0. 01);BALF 中 IL-1β、IL-8、TNF-α 含量明显升高(P< 0. 01), MDA 含量显著升高(P< 0. 01),SOD 含量显著降低(P< 0. 01);HE 染色可见明显肺泡间隔增厚、肺间质水肿、红细胞渗出。 结论 暴露式气管滴注 LPS 法,可以引起大鼠肺功能显著下降、剧烈的肺部炎症、氧化-抗氧化失衡及严重肺水肿和肺出血,导致急性肺损伤,在 3 h 时段更有利于 ALI 大鼠模型的构建。
英文摘要:
       Objective A rat model of acute lung injury (ALI) was established by intratracheal instillation of lipopolysaccharide (LPS) and changes in the lung injury were observed at various periods. Methods Thirty-two healthy Sprague-Dawley (SD) rats were randomly divided into normal (n= 8) and model (n= 24) group. In accordance with the duration of LPS infusion, the model group was divided into three subgroups: 3, 6 and 12 h groups, with eight rats in each group. The ALI rat model was established by tracheal instillation of LPS ( 2 mg / kg). Observations included the general performance of rats, gross observation of lungs, detection of lung functions, calculation of the lung wet / dry weight ratio, detection of interleukin ( IL) - 1β, IL-8, and tumor necrosis factor - α in bronchoalveolar lavage fluid, detection of malondialdehyde and superoxide dismutase in lung tissue, and histomorphological observation by hematoxylin-eosin staining in lung tissue. Changes of acute lung injury were also evaluated at different stages. Results After modeling, the survival rate of rats in the model group was 100%. Compared with the normal group, the general performances of rats in the 3 h group were similar with less food intake, less activity, more mucus secretions in the nasal cavity, faster respiratory frequency, and audible wheezing. Gross observation of the lungs showed liver-like degeneration of the lung tissue in the left and right hilum of the lung in the 3 h group, bleeding spots were scattered on the left and right lobes, and the bleeding site was bright red. The pulmonary functions of rats after LPS exposure for 3 h showed significant decreases in the forced expiratory volume (FEV) in 0. 1 s, forced expiratory volume in 0. 3 s, ratio of forced expiratory volume in 0. 1 s to forced vital capacity, and ratio of forced expiratory volume in 0. 3 s to forced vital capacity (P< 0. 05,P<0. 01). The wet / dry weight ratio was increased significantly ( P< 0. 01). The contents of IL-1β, IL-8, and tumor necrosis factor - α in bronchoalveolar lavage fluid were increased significantly ( P< 0. 01). The content of malondialdehyde was increased significantly (P< 0. 01). The content of superoxide dismutase was decreased significantly (P<0. 01). Hematoxylin-eosin staining showed obvious thickening of the alveolar septum, pulmonary interstitial edema, and erythrocyte exudation. Conclusions Tracheal instillation of LPS causes significant decreases in lung functions, severe pulmonary inflammation, an oxidation-antioxidation imbalance, and severe pulmonary edema in rats, which lead to acute lung injury. Furthermore, it is more beneficial to establish an ALI rat model at the 3 h time point.
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