Airway Research, Fraunhofer Institute for Toxicology and Experimental Medicine, Hannover, Germany.
PLoS One. 2012;7(8):e43709. doi: 10.1371/journal.pone.0043709. Epub 2012 Aug 28.
Increasing incidence and substantial morbidity and mortality of respiratory diseases requires the development of new human-specific anti-inflammatory and disease-modifying therapeutics. Therefore, new predictive animal models that closely reflect human lung pathology are needed. In the current study, a tiered acute lipopolysaccharide (LPS)-induced inflammation model was established in marmoset monkeys (Callithrix jacchus) to reflect crucial features of inflammatory lung diseases. Firstly, in an ex vivo approach marmoset and, for the purposes of comparison, human precision-cut lung slices (PCLS) were stimulated with LPS in the presence or absence of the phosphodiesterase-4 (PDE4) inhibitor roflumilast. Pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-α) and macrophage inflammatory protein-1 beta (MIP-1β) were measured. The corticosteroid dexamethasone was used as treatment control. Secondly, in an in vivo approach marmosets were pre-treated with roflumilast or dexamethasone and unilaterally challenged with LPS. Ipsilateral bronchoalveolar lavage (BAL) was conducted 18 hours after LPS challenge. BAL fluid was processed and analyzed for neutrophils, TNF-α, and MIP-1β. TNF-α release in marmoset PCLS correlated significantly with human PCLS. Roflumilast treatment significantly reduced TNF-α secretion ex vivo in both species, with comparable half maximal inhibitory concentration (IC(50)). LPS instillation into marmoset lungs caused a profound inflammation as shown by neutrophilic influx and increased TNF-α and MIP-1β levels in BAL fluid. This inflammatory response was significantly suppressed by roflumilast and dexamethasone. The close similarity of marmoset and human lungs regarding LPS-induced inflammation and the significant anti-inflammatory effect of approved pharmaceuticals assess the suitability of marmoset monkeys to serve as a promising model for studying anti-inflammatory drugs.
呼吸道疾病发病率不断上升,且发病率和死亡率高,这就需要开发新的、针对人类的抗炎和疾病修饰治疗方法。因此,需要开发新的、能够更紧密反映人类肺部病理的预测性动物模型。在本研究中,我们建立了一个分层急性脂多糖(LPS)诱导的炎症模型,以反映炎症性肺部疾病的关键特征。首先,我们采用离体方法,用 LPS 刺激食蟹猴(Callithrix jacchus)和人离体肺切片(PCLS),并在有无磷酸二酯酶-4(PDE4)抑制剂罗氟司特的情况下进行刺激。我们测量了包括肿瘤坏死因子-α(TNF-α)和巨噬细胞炎症蛋白-1β(MIP-1β)在内的促炎细胞因子。地塞米松作为皮质类固醇治疗对照。其次,在体内方法中,我们用罗氟司特或地塞米松预处理食蟹猴,然后用 LPS 对其进行单侧处理。在 LPS 处理后 18 小时,对同侧支气管肺泡灌洗(BAL)进行处理和分析。BAL 液经处理后,用于分析中性粒细胞、TNF-α和 MIP-1β。食蟹猴 PCLS 中的 TNF-α释放与人类 PCLS 显著相关。罗氟司特在两种物种中均能显著减少体外 TNF-α的分泌,半数最大抑制浓度(IC50)相似。LPS 注入食蟹猴肺部会导致炎症反应,表现为中性粒细胞浸润以及 BAL 液中 TNF-α和 MIP-1β水平升高。罗氟司特和地塞米松能显著抑制这种炎症反应。LPS 诱导的炎症在食蟹猴和人类肺部具有相似性,以及已批准药物的显著抗炎作用,评估了食蟹猴作为研究抗炎药物的有前途模型的适用性。