毒素清对肺炎克雷伯杆菌肺炎老龄大鼠心肌损伤中炎症细胞因子表达的影响

Effect of Dusuqing(毒素清) on the inflammatory cytokine in aged rats′ heart injury of Klebsiella pneumoniae

目的从心肌组织肿瘤坏死因子-α(TNF-α)、白细胞介素-1(IL-1)、巨噬细胞炎症蛋白-2(MIP-2)蛋白表达和MIP-2 mRNA表达变化揭示中药毒素清对老年肺炎心肌损伤的保护作用机制。方法将老龄大鼠随机分为对照组、模型组、毒素清组和左氧氟沙星组,制备肺炎克雷伯杆菌肺炎模型。计数外周血和支气管肺泡灌洗液(BALF)中的白细胞(WBC)计数和中性粒细胞(PMN)计数,采用放射免疫法测定外周血IL-1、TNF-α水平,采用免疫组化方法和原位杂交方法结合图像分析技术测定心肌组织MIP-2、IL-1、TNF-α蛋白表达及MIP-2 mRNA表达。结果与对照组比较,模型组外周血和BALF中WBC、PMN明显升高,血清和心肌组织IL-1、TNF-α水平明显增加,MIP-2的蛋白和mRNA表达明显增强(P<0.05或P<0.01);与模型组比较,左氧氟沙星组和毒素清组外周血和BALF中WBC、PMN明显减少,血清和心肌组织IL-1、TNF-α水平降低,MIP-2的蛋白和mRNA表达明显减弱(P均<0.01)。结论细胞因子TNF-α、IL-1、MIP-2紊乱参与老年肺炎导致心肌损伤的发生发展。阻抑TNF-α、IL-1、MIP-2介导的损伤可能是毒素清保护老年肺炎时心肌损伤的主要途径。

Objective To study traditional Chinese medicine Dusuqing＇s （毒素清） mechanism of protecting heart injury in aged Klebsiella pneumoniae by the changes of tumor necrosis faetor-α （TNF-α）, interleukin-1 （IL-1）, maerophage inflammatory protein-2 （MIP-2） and MIP-2 mRNA. Methods The rats were randomly divided into control group, model group, Dusuqing group, levofloxaein group, and the rat model of Klebsiella pneumoniae was established. The white blood cell （WBC） and polymorphonuelear leucocyte （PMN） count in peripheral blood and bronehoalveolar lavage fluid （BALF） were counted, radio-immunological method was applied to determine the levels of IL-1, TNF-α in peripheral blood, and immunohistoehemistry, hybridization in situ and image analysis were used to observe MIP-2, IL-1, TNF-α protein expressions and MIP-2 mRNA expression in the cardiac muscle. Results Compared with the control group, the counts of WBC and PMN in peripheral blood and BALF were markedly increased and the levels of serum and myocardial IL-1, TNF-α, MIP-2 protein and MIP-2 mRNA were strengthened in the model group （P〈0.05 or P〈0. 01）. Compared with the model group, the counts of WBC and PMN in peripheral blood and BALF were decreased significantly, the levels of serum and myocardial IL-1, TNF-α were lowered and MIP-2 protein and MIP-2 mRNA expressions were weakened in the Dusuqing and levofloxaein groups （all P〈 0.01）. Conclusion The disorders of eytokines, TNF-α, IL-1, MIP-2 are involved in the occurrence and development of myocardial damage induced by the aged pneumonia. The suppression of TNF-α, IL-1 and MIP-2 mediated injury is possibly the main pathway of Dusuqing for the protection of myocardial damage in aged pneumonia.