BML-111通过抑制MAPK信号通路减轻大鼠失血性休克所致肺损伤

发布时间：2019-02-13 20:44

【摘要】：目的：探讨脂氧素受体激动剂（lipoxin receptor agonist）BML-111对失血性休克（hemorrhagic shock,HS）所致大鼠急性肺损伤（acute lung injury,ALI）的影响及其作用机制。 方法：选择32只健康雄性SD大鼠，根据是否放血以及给药的不同随机分为4组，分别为对照(sham)组，失血性休克/复苏(HS)组，BML-111干预(BML-111)组，BMl-111加BOC-2干预（BOC-2）组。将大鼠麻醉后依次进行左侧颈总动脉插管与右侧颈静脉插管。左侧动脉插管连接压力转换器，用以放血和检测平均动脉压（mean arterialpressure，MAP）的变化。右侧颈静脉插管用于补液复苏。除sham组大鼠外，其余三组大鼠均进行放血处理，使其失去35%血容量造成失血性休克模型，休克持续30分钟后进行复苏。BML-111组复苏开始时腹腔注射1mg/kg剂量的BML-111，BOC-2组在麻醉后腹腔注射50μg/kg剂量的BOC-2，并于复苏开始时腹腔注射1mg/kg剂量的BML-111。所有组大鼠在复苏结束后2小时放血处死，收集支气管肺泡灌洗液（bronchoalveolar lavage fluid,BALF）并取肺组织标本于-80℃冰箱保存。HE切片染色观察各组大鼠标本的肺组织损伤情况；检测肺湿干重比（W/D）；检测BALF细胞分类计数；免疫组织化学染色法检测髓过氧化物酶（myeloperoxidase，MPO）表达水平；染色酶联免疫法（enzyme-linked immunosorbent assay, ELISA）检测肺组织IL-1β和IL-6表达水平；免疫蛋白印迹法（western-blot assay，WB）检测大鼠肺组织丝裂原活化蛋白激酶（mitogen activated protein kinase，MAPK）磷酸化激活水平；电泳迁移率分析法（electrophoretic mobility shift assays，EMSA）检测激活蛋白1（activator protein-1，AP-1）与DNA的结合活性。 结果：（1）显微镜观察结果显示sham组大鼠肺组织切片无明显炎症损伤改变；HS组肺组织切片有明显的肺组织炎性细胞浸润，肺泡壁增厚，透明膜形成等炎症损伤表现；BML-111组炎症损伤明显减轻；BOC-2组则表现出与HS组相似的病理组织学改变。（2）相对于sham组，HS组BALF中性粒细胞计数明显增高（P0.05）；相对于HS组，BML-111组BALF中性粒细胞计数明显减少（P0.05）；相对于BML-111组，BOC-2组BALF中性粒细胞计数明显增高（P0.05），与HS组数量相近。（3）与sham组相比，HS组肺组织W/D明显升高（P0.05）；与HS组相比，BML-111组肺组织W/D明显下降（P0.05）；在对大鼠BOC-2干预后，肺组织W/D又明显升高（P0.05）。（4）与sham组相比，HS组MPO阳性表达明显增多（P0.05）；与HS组相比，BML-111组MPO阳性表达显著下降（P0.05）；与BML-111组相比，BOC-2组MPO阳性表达量又明显升高（P0.05）。（5）相对于sham组，HS组肺组织IL-1β和IL-6含量显著增多（P0.05）；相对于HS组，BML-111使大鼠肺组织IL-1β和IL-6含量明显下降（P0.05；使用BOC-2后，大鼠肺组织IL-1β和IL-6含量相比于BML-111组又明显增多（P0.05。（6）与sham组相比，HS组大鼠肺组织ERK，JNK，p38MAPK磷酸化水平明显提高（P0.05）；使用BML-111使大鼠肺组织ERK，JNK，p38MAPK磷酸化水平相较于HS组明显降低（P0.05）；而在对大鼠注射BOC-2后，ERK，JNK，p38MAPK磷酸化水平相比于BML-111组又明显升高（P0.05）。（7）与sham组相比，HS组AP-1DNA结合活性显著升高；与HS组相比，，BML-111组AP-1DNA结合活性明显下降；BOC-2组AP-1DNA结合活性相对于BML-111组显著提高。 结论：BML-111可通过抑制MAPK/AP-1信号通路的激活减轻失血性休克引起的大鼠急性肺损伤炎症反应。
[Abstract]:Objective: To study the effect of BML-111 (BML-111) on acute lung injury (ALI) in rats with hemorrhagic shock (HS) and its mechanism of action. Methods: 32 healthy male SD rats were randomly divided into 4 groups according to whether exsanguination and administration were divided into 4 groups: sham group, hemorrhagic shock/ resuscitation (HS) group, BML-111 intervention (BML-111) group, BMl-111 plus BOC-2 intervention (BOC-2). Group. After the rats were anesthetized, the left common carotid artery and the right jugular vein were performed in turn The left-hand arterial cannula is connected to a pressure transducer for bleeding and detecting a change in the mean arterial pressure (MAP) The right jugular vein is used for rehydration. In addition to the sham group, the rest of the rats were exsanguinated to lose 35% of the blood volume, resulting in the hemorrhagic shock model, and the shock lasted for 30 minutes. The BML-111, BOC-2 group at the dose of 1 mg/ kg was injected intraperitoneally at the beginning of the resuscitation at the beginning of the resuscitation, and the dose of 1 mg/ kg of BML-11 was injected into the abdominal cavity at the beginning of the resuscitation. 1. All the rats were exsanguinated for 2 hours after the end of the resuscitation, and the bronchoalveolar lavage fluid (BALF) was collected and the lung tissue specimens were taken from the refrigerator at-80 鈩

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