中和IL-17A对输血相关急性肺损伤小鼠的保护作用

Neutralisation of IL-17A protect mice in a two-event model of transfusion-related acute lung injury

目的：观察IL-17A在输血相关急性肺损伤小鼠中的表达，探讨中和IL-17A对输血相关急性肺损伤小鼠的影响。方法采用“二次打击”（LPS＋MHC-ⅠmAb）模型，8～10周BALB/C雄性小鼠32只，应用随机数据表完全随机分成4组：正常（normal）组、LPS＋MHC-ⅠmAb组、LPS＋isotype组、LPS＋PBS组，每组8只。腹腔注射LPS 0.1 mg/kg，24 h后尾静脉分别给予MHC-ⅠmAb（1 mg/kg）或等体积的isotype和PBS，2 h后检测肺泡灌洗液和外周血中IL-17的表达。另选取32只BALB/C雄性小鼠，观察anti-IL-17A抗体预处理对肺损伤的影响，随机分为4组：normal 组、LPS＋MHC-ⅠmAb＋anti-IL-17A 组、LPS＋MHC-ⅠmAb＋isotype 组、LPS＋MHC-ⅠmAb＋PBS组，提前1 h尾静脉注射anti-IL-17A（50μg/只）、isotype（50μg/只）或等体积的PBS，二次打击造模后2 h取材，测定肺泡灌洗液中蛋白浓度和肺干湿重比，检测肺泡灌洗液和外周血中细胞因子水平，计数肺脏中性粒细胞，评估各组小鼠肺损伤和炎症反应程度。数据采用 Prism5.0（GraphPad Software，USA）软件包进行统计学处理。结果与其他三种相比，二次打击模型（LPS＋MHC-ⅠmAb）组小鼠肺泡灌洗液和外周血中IL-17A表达显著升高，且肺组织中性粒细胞数明显增多。anti-IL-17A预处理后，肺组织中性粒细胞浸润减少，肺干湿重比减小，肺泡灌洗液蛋白含量降低，外周血促炎因子水平显著下调。结论 IL-17A在输血相关急性肺损伤中显著升高，anti-IL-17A预处理后显著改善输血相关急性肺损伤小鼠炎症反应和肺组织损伤。

ObjectiveTo study the effect of IL-17A in mice model of transfusion-related acute lung injury and to determine that anti-IL-17A may alleviate the lung injury.MethodsTwo-hit model was used in the study. 32 BALB/C male mice were randomly assigned to 4 groups： normal control group, LPS＋MHC-ⅠmAb group, LPS＋isotype group, LPS＋PBS group. ALI was induced intraperitoneally injection of LPS （1 mg/kg）. After 24 h, the animals were intravenously given injection of MHC-ⅠmAb, isotype and PBS. And mice were killed after 2 h. The concentration of IL-17A was detected in the peripheral blood and BALF, the lung wet/dry weight ratio, TP level, and the lung pathologic tissue score with HE strain. Secondly to evaluate whether anti-IL-17 antibody can alleviate the lung injury. BALB/C male mice were injected with anti-IL-17 antibody, isotype and PBS by tail intraveous. After 1 h, mice received intraperitoneally injection of LPS, then after 24 h all were intravenously given injection of MHC-ⅠmAb, then divided into anti IL-17 group, isotype＋MHC-Ⅰ group and the PBS＋MHC-Ⅰ group. And mice were killed after 2 hours. ALI markers and inflammatory factors, including lung wet/dry weight ratio, cytokines level, and the lung pathologic damage with HE strain, were measured in our experiment.ResultsCompared to others, the expression of IL-17A in BALF and peripheral blood in two-event model markedly increased significantly. The protein level increased in BALF, ALI aggravated. With the preprocessing of anti-IL-17A antibody, IL-17A decreased, the lung wet/dry weight ratio grew down. The protein level decreased in BALF, inflammatory factors decreased significantly.ConclusionsIL-17A upregulated in TRALI and aggravated lung injury. The neutralization of IL-17A can alleviate acute lung injury and inhibit the inflammatory response.