摘要目的 研究输血相关急性肺损伤(TRALI)大鼠肺组织核转录因子(NF)-κB及细胞间黏附分子(ICAM)-1表达变化,并探讨其变化的临床意义.方法 60只SD大鼠按随机数字表法分为正常对照组、脂多糖对照组、TRALI组和阳性对照组各15只.正常对照组:采用假手术处理;脂多糖对照组:经大鼠腹腔注射脂多糖(2 mg/kg,≤1 min完毕),2 h后静脉输注生理盐水(约1 ml/只);TRALI组:腹腔注射脂多糖(2 mg/kg)2 h后静脉输注血浆(约1 ml/只);阳性对照组:静脉输注脂多糖(5 mg/kg)诱导急性肺损伤.支气管肺泡灌洗液(BALF)沉渣行瑞士染色进行白细胞分类计数;大鼠肺组织病变进行半定量肺损伤评分;采用免疫组织化学法对肺泡上皮细胞中ICAM-1表达进行定位及半定量分析,并对NF-κB核转位进行半定量分析.结果 阳性对照组和TRALI组大鼠支气管肺泡灌洗液多形核白细胞(PMN)比例[(39.6±12.5)％和(33.5±11.5)％]及肺损伤评分[(2.32±0.70)和(1.98±0.87)]均明显高于正常对照组[(9.8±3.5)％和(0.00±0.00)]和脂多糖对照组[(11.4±4.1)％和(1.07±0.48)],(均P<0.05);阳性对照组和TRALI组大鼠肺泡上皮细胞中ICAM-1蛋白[(0.28±0.09)和(0.24±0.07)]和ICAM-1 mRNA表达[(1.53±0.17)和(1.24±0.11)]明显高于正常对照组[(0.12±0.03)和(0.36±0.05)]和脂多糖对照组[(0.16±0.07)和(0.89±0.10)](均P<0.05);阳性对照组和TRALI组大鼠肺泡上皮细胞中NF-κB核表达[(23.8±3.20)和(27.4±4.60)]明显高于正常对照组(6.90±2.30)和脂多糖对照组(11.7±2.80),(均P<0.05).结论 TRALI大鼠肺组织中ICAM-1蛋白和ICAM-1 mRNA的表达升高,肺组织PMN浸润增加,同时大鼠肺泡上皮细胞中NF-κB核表达升高,提示ICAM-1可能参与了PMN与内皮细胞的黏附和聚集,推测NF-κB激活可能参与了TRALI炎症介质ICAM-1基因转录调控,在TRALI的病理生理过程中可能发挥重要作用.

abstractsObjective To determine the change in the expression of nuclear factor-κB(NF-κB) and intercellular adhesion molecule 1(ICAM-1)in the lung tissues of rats with transfusion-related acute lung injury,and to investigate its clinical significance. Methods Sixty SD rats were randomly divided into the normal control group,lipopolysaccharide(LPS)control group,TRALI group and positive control group(n=15 each). The normal control group was treated with sham operation. The LPS control group was intraperitoneally injected with LPS (2 mg/kg,completed within ≤ 1 min),and was given intravenous infusion of normal saline at 2 h(about 1 ml/rat). The TRALI group was treated with intravenous infusion of blood(about 1 ml/rat)at 2 h after intraperitoneal injection of LPS(2 mg/kg). The positive control group was treated with intravenous infusion of LPS(5 mg/kg)to induce acute lung injury. The sediment of bronchial alveolar lavage fluid(BALF)was determined by Wright-Giemsa staining for differential leukocyte count. The lesions in the lung tissue of rats were assessed by semi-quantitative lung injury scoring. The expression of ICAM-1 in alveolar epithelial cells was determined by immunohistochemistry and semi-quantitative analysis. The NF-κB nuclear translocation was determined by semi-quantitative analysis. Results In the positive control group and the TRALI group,the ratio of polymorphonuclear leukocytes(PMN)in the bronchoalveolar lavage fluid[(39.6±12.5)％and(33.5±11.5)％]and the lung injury score[(2.32±0.70)and(1.98±0.87)] were significantly higher than those in the normal control group[(9.8±3.5)％and(0.00±0.00)]and the LPS control group[(11.4±4.1)％and(1.07±0.48)],respectively(all P<0.05). In the positive control group and TRALI group,the expressions of ICAM-1 protein[(0.28 ± 0.09)and(0.24 ± 0.07)]and ICAM-1 mRNA [(1.53 ± 0.17)and(1.24 ± 0.11)]in the alveolar epithelial cells were significantly higher than those in the normal control group[(0.12 ± 0.03)and(0.36 ± 0.05)]and LPS control group[(0.16 ± 0.07)and(0.89 ± 0.10)],respectively(all P<0.05). In the positive control group and TRALI group,the expression of NF-κB in the alveolar epithelial cells[(23.8 ± 3.20)and(27.4 ± 4.60)]was significantly higher than that in the normal control group(6.90 ± 2.30)and the LPS group(11.7 ± 2.80),respectively(all P<0.05). Conclusion In the lung tissues of TRALI rats,the expressions of ICAM-1 protein and ICAM-1 mRNA,and the infiltration of PMN increase. Meanwhile,the expression of NF-κB increases in rat alveolar epithelial cells, indicating that ICAM-1 may be involved in adhesion and aggregation of PMN and endothelial cells, suggesting that NF-κB activation may be involved in the regulation of TRALI inflammatory mediators and ICAM-1 gene transcription,which may play a significant role in the pathophysiology of TRALI.