【摘要】：目的：探討脂氧素受體激動(dòng)劑（lipoxin receptor agonist）BML-111對(duì)失血性休克（hemorrhagic shock,HS）所致大鼠急性肺損傷（acute lung injury,ALI）的影響及其作用機(jī)制。 方法：選擇32只健康雄性SD大鼠，根據(jù)是否放血以及給藥的不同隨機(jī)分為4組，分別為對(duì)照(sham)組，失血性休克/復(fù)蘇(HS)組，BML-111干預(yù)(BML-111)組，BMl-111加BOC-2干預(yù)（BOC-2）組。將大鼠麻醉后依次進(jìn)行左側(cè)頸總動(dòng)脈插管與右側(cè)頸靜脈插管。左側(cè)動(dòng)脈插管連接壓力轉(zhuǎn)換器，用以放血和檢測平均動(dòng)脈壓（mean arterialpressure，MAP）的變化。右側(cè)頸靜脈插管用于補(bǔ)液復(fù)蘇。除sham組大鼠外，其余三組大鼠均進(jìn)行放血處理，使其失去35%血容量造成失血性休克模型，休克持續(xù)30分鐘后進(jìn)行復(fù)蘇。BML-111組復(fù)蘇開始時(shí)腹腔注射1mg/kg劑量的BML-111，BOC-2組在麻醉后腹腔注射50μg/kg劑量的BOC-2，并于復(fù)蘇開始時(shí)腹腔注射1mg/kg劑量的BML-111。所有組大鼠在復(fù)蘇結(jié)束后2小時(shí)放血處死，收集支氣管肺泡灌洗液（bronchoalveolar lavage fluid,BALF）并取肺組織標(biāo)本于-80℃冰箱保存。HE切片染色觀察各組大鼠標(biāo)本的肺組織損傷情況；檢測肺濕干重比（W/D）；檢測BALF細(xì)胞分類計(jì)數(shù)；免疫組織化學(xué)染色法檢測髓過氧化物酶（myeloperoxidase，MPO）表達(dá)水平；染色酶聯(lián)免疫法（enzyme-linked immunosorbent assay, ELISA）檢測肺組織IL-1β和IL-6表達(dá)水平；免疫蛋白印跡法（western-blot assay，WB）檢測大鼠肺組織絲裂原活化蛋白激酶（mitogen activated protein kinase，MAPK）磷酸化激活水平；電泳遷移率分析法（electrophoretic mobility shift assays，EMSA）檢測激活蛋白1（activator protein-1，AP-1）與DNA的結(jié)合活性。 結(jié)果：（1）顯微鏡觀察結(jié)果顯示sham組大鼠肺組織切片無明顯炎癥損傷改變；HS組肺組織切片有明顯的肺組織炎性細(xì)胞浸潤，肺泡壁增厚，透明膜形成等炎癥損傷表現(xiàn)；BML-111組炎癥損傷明顯減輕；BOC-2組則表現(xiàn)出與HS組相似的病理組織學(xué)改變。（2）相對(duì)于sham組，HS組BALF中性粒細(xì)胞計(jì)數(shù)明顯增高（P0.05）；相對(duì)于HS組，BML-111組BALF中性粒細(xì)胞計(jì)數(shù)明顯減少（P0.05）；相對(duì)于BML-111組，BOC-2組BALF中性粒細(xì)胞計(jì)數(shù)明顯增高（P0.05），與HS組數(shù)量相近。（3）與sham組相比，HS組肺組織W/D明顯升高（P0.05）；與HS組相比，BML-111組肺組織W/D明顯下降（P0.05）；在對(duì)大鼠BOC-2干預(yù)后，肺組織W/D又明顯升高（P0.05）。（4）與sham組相比，HS組MPO陽性表達(dá)明顯增多（P0.05）；與HS組相比，BML-111組MPO陽性表達(dá)顯著下降（P0.05）；與BML-111組相比，BOC-2組MPO陽性表達(dá)量又明顯升高（P0.05）。（5）相對(duì)于sham組，HS組肺組織IL-1β和IL-6含量顯著增多（P0.05）；相對(duì)于HS組，BML-111使大鼠肺組織IL-1β和IL-6含量明顯下降（P0.05；使用BOC-2后，大鼠肺組織IL-1β和IL-6含量相比于BML-111組又明顯增多（P0.05。（6）與sham組相比，HS組大鼠肺組織ERK，JNK，p38MAPK磷酸化水平明顯提高（P0.05）；使用BML-111使大鼠肺組織ERK，JNK，p38MAPK磷酸化水平相較于HS組明顯降低（P0.05）；而在對(duì)大鼠注射BOC-2后，ERK，JNK，p38MAPK磷酸化水平相比于BML-111組又明顯升高（P0.05）。（7）與sham組相比，HS組AP-1DNA結(jié)合活性顯著升高；與HS組相比，，BML-111組AP-1DNA結(jié)合活性明顯下降；BOC-2組AP-1DNA結(jié)合活性相對(duì)于BML-111組顯著提高。 結(jié)論：BML-111可通過抑制MAPK/AP-1信號(hào)通路的激活減輕失血性休克引起的大鼠急性肺損傷炎癥反應(yīng)。
[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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