创伤小鼠脾细胞核因子-κB的表达及与全身炎症反应综合征发生的关系

The relationship between activation of nuclear factor-κB and systemic inflammatory response syndrome in mice spleen lymphocytes after trauma

目的:探讨创伤后不同时相点脾细胞核因子-kappaB(NF-κB)的DNA结合蛋白和NF-κB P^(65)蛋白亚型表达的动态变化。方法:采用小鼠双后肢闭合性砸伤+骨折模型,于创伤后1、4、7d处死动物,分离、纯化脾细胞,提取脾细胞核蛋白。用电泳迁移率改变试验(electrophoretic mobility shift assay,EMSA)检测NF-κB的DNA结合活性,用免疫蛋白印变法(Western blotting)检测NF-κB P^(65)蛋白亚型表达。结果:NF-κB的DNA结合活性在创伤后1、4、7d表达明显增加。NF-κB P^(65)蛋白亚型在创伤后1、4、7d表达明显增高。结论:创伤可明显增强脾细胞NF-κB的DNA结合活性和P^(65)蛋白亚型的表达,在创伤后淋巴细胞活化及全身炎症反应综合征(SIRS)发生、发展过程中可能起重要作用。

Objective: To observe the dynamic changes in the activation of NF-κB and the expression of P^(65) protein subunits of NF-κB in mice spleen lymphocytes after trauma. Methods: Hind limbs closed impact injury combined with fracture in mice was adopted as the trauma model, and animals were sacrificed on days 1, 4 and 7 after injury. Spleen lymphocytes were isolated from traumatized mice, and nuclear extracts were obtained. The DNA binding activity of NF-κB was measured using electrophoretic mobility shift assay (EMSA). The expression of P^(65) protein subunits of NF-κB was determined by Western blotting. Results: The DNA binding activity of NF-κB was gradually elevated in traumatized mice spleen lymphocytes and significantly increased on days 1,4 and 7 after injury. Western blotting showed marked expression of P^(65) protein of NF-κB on days 1、4 and 7 after injury. Conclusions: These data suggested that the expression of the DNA binding activity and P^(65) protein subunits of NF-κB were significantly increased in spleen lymphocytes of traumatized mice, and the changes may play an important role in mediating lymphocytes activation caused by trauma and development of systemic inflammatory response syndrome (SIRS).