胸部开放伤后海水灌注致急性肺损伤犬模型前B细胞集落刺激因子水平的变化

Changes in the level of PBEF in acute lung injury induced by seawater perfusion following open chest injury

目的探讨胸部开放伤后胸腔海水灌注致急性肺损伤（ALI）早期前B细胞集落刺激因子（pre—B—cellcolony—enhancing factor，PBEF）水平的变化及可能作用。方法16只健康成年犬随机分为单纯胸部开放伤组（对照组）和伤后胸腔海水灌注组（海水组），于胸部开放伤后海水灌注前（0h）、灌注后6h取动、静脉血、支气管肺泡灌洗液（broncho—alveolar lavage fluid，BALF）和肺组织标本。测血浆渗透压和肺泡通透指数；酶联免疫吸附试验测血浆、BALF及肺组织匀浆中、PBEF的含量；Western bloting方法测肺组织PBEF蛋白浓度，Real—time reverse PCR方法测肺组织中编码PBEF mRNA表达。结果海水组动物PBEF蛋白[（3014．16±883．47）ng／L]明显高于对照组[（1060．94±251．08）ng／L]；Western bloting量化值比较：海水组为（0．233±0．037）ng／L，对照组为（0．154±0．013）ng／L，差异有统计学意义（P〈0．05）；PBEFmRNA：量化相对值海水组为5．27±0．05，对照组为0．65±0．12，差异有统计学意义（P〈0．05）；血浆PBEF浓度明显升高，与血浆渗透压明显相关（r=0．744，P〈0．05）。结论胸部开放伤后胸腔海水灌注致ALI早期PBEF升高，与海水高渗有关；PBEF参与ALI病理生理过程，可能作为胸部开放伤合并胸腔海水灌注致ALI早期较为敏感的监测指标之一。

Objective To investigate changes in the level of pre-B-cell colony-enhancing factor （PBEF） and its possible effect on acute lung injury （ALI） induced by seawater perfusion following open chest injury in dogs. Methods Sixteen healthy adult dogs were randomly divided into the pure open chest injury group （the control group） and seawater perfusion after injury group （later simply referred to as the seawater group）. Plasma, bronchoalveolar lavage fluid （BALF） and lung tissue samples were collected before seawater- perfusion （0 h） and 6 h after seawater perfusion. The PBEF protein levels in plasma, BALF and lung tissue were measured with enzyme-linked immunosorbent assay （ELISA） , immunohistochemistry and Western blot. The expression of mRNA synthesis of coding PBEF in lung homogenates was evaluated with real-time RT-PCR. Results Compared with those of the control animals, PBEF levels significantly increased in the animals of the seawater group [plasma：（3014.16±883.47） ng/L versus （1060.94±251.08） ng/L （P〈0.05）; lung tissue（ Western blot） ： （0. 233±0. 037 ） versus （0. 1541±0. 013 ） ng/L, with statistical signifieanee（ P 〈 0.05） ;PBEF mRNA ：5.27±0.05 versus 0.65±0. 12, with obvious statistical signifieance（ P 〈 0.05 ） 1. PBEF levels were positively eorrelated with plasma osmotic pressure, with a relative coefficient of 0. 744 （P 〈 0.05 ）. Conclusions Seawater perfusion could elevate the level of PBEF in animals with ALl induced by seawater perfusion following open chest injury, indicating that it might be closely related with the high permeability of seawater. PBEF was clearly involved in the pathophysiological process of ALI, and might be a novel biomarker which could be used to identify ALl induced by seawater perfusion.