TRALI大鼠肺泡活化巨噬细胞及共刺激分子CD40、ICAM-1的表达变化分析

摘要

目的 探讨输血相关急性肺损伤(TRALI)大鼠肺泡活化巨噬细胞及共刺激分子CD40、血浆及肺组织中细胞间粘附分子-1(ICAM-1)的表达变化及其意义.方法 选取60只SPF级、6-7周龄的雄性SD大鼠,采用随机数字表法分为假手术组(腹腔注射等量生理盐水)、对照组(腹腔注射2mg/kg脂多糖)、模型组(静脉输注脂多糖5mg/kg)和TRALI组(腹腔注射2mg/kg脂多糖+静脉输注血浆1mL/只)各15只,采用RT-PCR技术检测活化巨噬细胞中Toll样受体4(TLR4)mRNA的表达,采用Northern bolt 检测CD40 mRNA的表达,采用免疫组化染色检测ICAM-1蛋白的表达,采用酶联免疫吸附法(ELISA)检测血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-β)、ICAM-1的水平.结果 模型组和TRALI组的活化巨噬细胞中TLR4 mRNA、肺组织中CD40 mRNA、ICAM-1蛋白表达水平显著高于假手术组和对照组(P<0.05),TRALI组的活化巨噬细胞中TLR4 mRNA、肺组织中CD40 mRNA、ICAM-1蛋白表达水平显著的低于模型组(P<0.05);模型组和TRALI组的血清中TNF-ɑ、IL-β、ICAM-1的水平显著的高于假手术组和对照组(P<0.05),TRALI组的血清中TNF-ɑ、IL-β、ICAM-1的水平显著的低于模型组(P<0.05).结论 TRALI大鼠肺TLR4 mRNA、CD40 mRNA、ICAM-1蛋白高表达,这可能与促进炎症因子释放,引起肺损伤有关.%Objective To investigate the expression and significance of ICAM-1 in CD40, plasma and lung tissue of rat alveolar activated macrophages and co-stimulatory molecules TRALI.Methods The value of 60 SPF, 6 to 7 week old male SD rats were randomly divided into the sham operation group (intraperitoneal injection of normal saline), the control group (intraperitoneal injection of lipopolysaccharide 2mg/kg), the model group (intravenous infusion of lipopolysaccharide 5mg/kg) and the TRALI group (intraperitoneal injection of 2mg/kg lipopolysaccharide+intravenous infusion of plasma 1ml/only), 15 rats in every group.The expression of TLR4 mRNA in macrophage activation was detected by RT-PCR technology, CD40 mRNA expression by Northern bolt, ICAM-1 protein by immunohistochemical staining, and the serum TNF-ɑ,IL-β and ICAM-1 levels by ELISA.Results The activation of lung tissue macrophages in TLR4 mRNA, CD40 mRNA, ICAM-1 protein expression level was significantly higher in the model group and the TRALI group than in the sham operation group and the control group (P<0.05).The expression levels of CD40 mRNA and ICAM-1 protein in lung tissue and TLR4 mRNA in the TRALI group were significantly lower than those in the model group (P<0.05).The levels of TNF-α, ICAM-1 and IL-β level were significantly higher in the model group and the TRALI group than in the sham group and the control group (P<0.05).The levels of TNF-α, ICAM-1 and IL-β level in serum were significantly lower in the TRALI group than in the model group (P<0.05).Conclusion High expression of TLR4 mRNA, CD40 mRNA and ICAM-1 protein in the lung of TRALI rats may be related with the promotion of the release of inflammatory factors and cause lung injury.