前B细胞克隆增强因子对急性肺损伤／急性呼吸窘迫综合征大鼠肺组织细胞黏附分子的影响

The influence of pre-B-cell colony enhancing factor on adhesive molecule in pulmonary cells in rats with acute lung injury/acute respiratory distress syndrome

目的观察前B细胞克隆增强因子（PBEF）对急性肺损伤，急性呼吸窘迫综合征（ALI／ARDS）大鼠肺组织细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）的影响。方法将40只SD大鼠按随机数字表法分为对照组、模型组、药物干预组、溶媒对照组，每组10只。采用油酸尾静脉注射复制ALI／ARDS模型。药物干预组制模前腹腔内注射PBEF抑制剂FK866，溶媒对照组制模前注射等体积FK866溶媒二甲亚砜。制模成功6h后取材，采用酶联免疫吸附试验（ELISA）测定支气管肺泡灌洗液（BALF）中肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）含量；观察肺组织病理学变化；用逆转录-聚合酶链反应和免疫组化染色测定肺组织中PBEF、ICAM-1、VCAM-1的mRNA及蛋白表达。结果与对照组比较，模型组大鼠肺组织出现明显的ALI／ARDS病理学改变，BALF中TNF-α（ng/L）、IL-1β（ng／L）含量增多（TNF-α：656．51±47．13比84．82±7．84，IL-1β：379．60±31．55比74．56±8．51，均P〈0．01），PBEF、ICAM-1、VCAM-1的mRNA和蛋白表达增加（PBEF mRNA：0．581±0．079比0．186±0．051，ICAM-1mRNA：0．558±0．060比0．176±0．070，VCAM-1mRNA：0．646±0．059比0．226±0．047；PBEF蛋白：0．089±0．024比0．037±0．011，ICAM-1蛋白：0．061±0．012比0．025±0．008，VCAM-1蛋白：0．072±0．013比0．033±0．010，均P〈0．01）。与模型组比较，药物干预组大鼠肺组织病理改变有所减轻，BALF中TNF-α、IL-1β含量显著降低（TNF-α：478．80±72．93比656．51±47．13，IL-1β：244．62±52．17比379．60±31．55，均P〈0．05），PBEF、ICAM-1、VCAM-1的mRNA和蛋白表达明显减少（PBEFmRNA：0．456±0．110比0．581±0．079，ICAM-1mRNA：0．413±0．073比0．558±0．060，VCAM-1mRNA：0．483±0．062比0．646±0．059；PBEF蛋白：0．059±0．010比0．089±0．024，ICAM-1蛋白：0．043±0．007比0．061±0．012，VCAM-1蛋白：0．050±0．009比0．072±0．013，均P〈0．05）。结论在ALI／ARDS发生时PBEF能通过增加黏附分子ICAM—1、VCAM-1表达促使炎陛细胞向肺内迁移和浸润，从而在其发生发展中起重要作用。

Objective To observe the influence of pre-B-cell colony enhancing factor （PBEF） on intercellular cell adhesion molecule-1 （ICAM-1） and vascular cell adhesion molecule-1 （VCAM-1） in lung tissue of rats with acute lung injury/acute respiratory distress syndrome （ALI/ARDS） induced by oleic acid. Methods A total of 40 male adult Sprague-Dawley （SD） rats were divided into control, model, drug intervention and vehicle control groups according to the random digits table with 10 rats in each group. ALI/ARDS was reproduced in the rats of model, drug intervention and vehicle control groups by injection of oleic acid （0.15 ml/kg） through the tail vein. The rats in drug intervention and vehicle control groups received the specific PBEF inhibitor FK866 （10 mg/kg）, while vehicle control group received the same volume of the vehicle only. Six hours after ALI/ARDS was successfully reproduced, bronchoalveolar alveolar lavage fluid （BALF） was obtained for the measurement of the contents of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） by enzyme linked immunosorbent assay （ELISA）. Lung tissue was obtained for pathological examination, and also for the measurement of the expression of PBEF, ICAM-1 and YCAM-1 mRNA by reverse transcription-polymerase chain reaction （RT-PCR）, and also the protein levels of PBEF, ICAM-1 and VCAM-1 by immunohistochemistry. Results Compared with rats in control group, the lung tissue of rats in model group showed distinctive pathological changes, the contents of TNF-α （ng/L） and IL-1β （ng/L） in BALF were increased （TNF-α 656.51 ± 47.13 vs. 84.82 ± 7.84, IL-113±379.60 ± 31.55 vs. 74.56 ± 8.51, both P〈0.01）, the mRNA and protein expression of PBEF, ICAM-1 and VCAM-1 were significantly increased （PBEF mRNA： 0.581 ± 0.079 vs. 0.186 ±0.051, ICAM-1 mRNA： 0.558 ±0.060 vs. 0.176 ±0.070, VCAM-1 mRNA： 0.646 ±0.059 vs. 0.226 ± 0.047 ; PBEF protein ： 0.089 ± 0.024 vs. 0.037 ± 0.011, ICAM-1 protein： 0.061 ± 0.012 vs. 0.025 ± 0.008, VCAM-1 protein： 0.072 ± 0.013 vs. 0.033 ±0.010, all P〈0.01）. Compared with model group, amelioration ofpathological change was found in lung tissue of rats in drug intervention group, the contents of TNF-oL and IL-1β in BALF were reduced （TNF-α 478.80 ± 72.93 vs. 656.51 ± 47.13, IL-β244.62 ± 52.17 vs. 379.60 ± 31.55, both P〈0.05）, and the mRNA and protein expression of PBEF, ICAM-1 and VCAM-1 were lowered （PBEF mRNA： 0.456 ± 0.110 vs. 0.581 ± 0.079, ICAM-1 mRNA： 0.413 - 0.073 vs. 0.558± 0.060, VCAM-1 mRNA： 0.483 ± 0.062 vs. 0.646 ± 0.059; PBEF protein： 0.059 ± 0.010 vs. 0.089 ± 0.024, ICAM-1 protein： 0.043 ± 0.007 vs. 0.061 ± 0.012, VCAM-1 protein： 0.050 ±0.009 vs. 0.072 ±0.013, all P〈0.05）. Conclusion PBEF could aggravate migration of pro-inflammatory cells to infiltrate the lung tissue by increasing the expression of ICAM-1 and VCAM-1, thus it plays an important role in the development of ALI/ARDS.