摘要
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.
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2013年第3期159-163,共5页
Chinese Critical Care Medicine
基金
基金项目:重庆市教委科研项目(KJ080323)