Vachon Eric, Bourbonnais Yves, Bingle Colin D, Rowe Sarah J, Janelle Marie France, Tremblay Guy M
Centre de Recherche, Hĵpital Laval, Institut Universitaire de Cardiologie et de Pneumologie de l'Université Laval, Quebec City, Canada.
Biol Chem. 2002 Jul-Aug;383(7-8):1249-56. doi: 10.1515/BC.2002.138.
The aim of the present study was to evaluate the anti-inflammatory activity of pre-elafin, an elastase-specific inhibitor, in lipopolysaccharide (LPS)-induced acute lung inflammation. C57BL/6 mice were pre-treated intranasally with recombinant human pre-elafin or vehicle only. One hour later, they were instilled intranasally with LPS (2 microg/mouse). Animals were sacrificed 6 hours after LPS instillation and bronchoalveolar lavage (BAL) was performed with three 1-ml aliquots of saline. LPS induced a lung inflammation characterised by a 100-fold increase in BAL neutrophils compared to control animals (265.8 +/- 54.5 x 10(3) and 2.4 +/- 1.3 x 10(3) neutrophils/ml, respectively). Pre-elafin dose-dependently reduced the neutrophil influx in the lung alveolar spaces by up to 84%. No elastase activity was detectable in all BAL fluids tested. Pre-elafin also reduced significantly LPS-induced gelatinase activity, as shown by zymography, and BAL macrophage inflammatory protein-2 (MIP-2) and KC levels, two potent neutrophil attractants and activators. Moreover, pre-elafin also significantly reduced mRNA levels of the three members of the IL-1 ligand family, namely IL-1alpha, IL-1beta and IL-1 receptor antagonist (IL-1Ra), type II IL-1 receptor, and TNFalpha as assessed in whole lung tissue by RNase protection assay. Thus, pre-elafin may be considered as a potent anti-inflammatory mediator.
本研究的目的是评估弹性蛋白酶特异性抑制剂前弹性蛋白在脂多糖（LPS）诱导的急性肺部炎症中的抗炎活性。将重组人前弹性蛋白或仅用赋形剂经鼻预处理C57BL/6小鼠。1小时后，经鼻向它们滴注LPS（2微克/只小鼠）。在滴注LPS后6小时处死动物，并用三份1毫升生理盐水进行支气管肺泡灌洗（BAL）。与对照动物相比，LPS诱导了肺部炎症，其特征是BAL中性粒细胞增加了100倍（分别为265.8±54.5×10³和2.4±1.3×10³个中性粒细胞/毫升）。前弹性蛋白剂量依赖性地将肺泡间隙中的中性粒细胞流入减少了高达84%。在所有测试的BAL液中均未检测到弹性蛋白酶活性。如酶谱分析所示，前弹性蛋白还显著降低了LPS诱导的明胶酶活性，以及BAL巨噬细胞炎性蛋白-2（MIP-2）和KC水平，这两种是有效的中性粒细胞趋化剂和激活剂。此外，通过核糖核酸酶保护试验在全肺组织中评估，前弹性蛋白还显著降低了IL-1配体家族三个成员，即IL-1α、IL-1β和IL-1受体拮抗剂（IL-1Ra）、II型IL-1受体和TNFα的mRNA水平。因此，前弹性蛋白可被视为一种有效的抗炎介质。
