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目的:从脏器组织Toll样受体4(toll-like receptor4,TLR4)信号转导相关分子表达变化方面揭示毒素清颗粒对老年细菌性肺炎大鼠多器官损伤的保护作用机制。方法:将55只老龄大鼠随机分为对照组、模型组、毒素清组和洛美沙星组,模型组25只,其余3组均为10只。气管插管,注入肺炎克雷伯杆菌,由肺炎导致多器官损伤;采用免疫组织化学法、逆转录聚合酶链反应法检测肺、心、小肠组织TLR4信号转导通路主要相关分子表达变化。结果:与对照组比较,模型组肺、心、小肠组织内毒素结合蛋白(lipopolysaccharide-binding protein,LBP)、CD14、TLR4、白细胞介素1受体相关激酶1(interleukin-1receptor-associated kinase-1,IRAK-1)mRNA和TLR4、肿瘤坏死因子受体相关因子6(tumor necrosis factor receptor-associated factor6,TRAF6)、核因子κB(nuclear factor-κB,NF-κB)蛋白表达明显增强(P<0.01,P<0.05)。与模型组比较,毒素清组肺、心、小肠组织LBP、CD14、TLR mRNA表达和TLR4、TRAF6、NF-κB蛋白表达显著减弱(P<0.01,P<0.05)。结论:毒素清可能通过降低TLR4信号转导活性,进而减少细胞因子分泌以减轻脏器组织损伤。毒素清可降低Toll样受体信号转导途径多种分子LBP、CD14、TLR4、IRAK-1、TRAF6和NF-κB活性,这与单一一种抑制剂仅作用于某个环节不同。
OBJECTIVE: To explore the protective mechanism of Tusuqing Granule on multiple organ injury in elderly bacterial pneumonia in the aspects of the changes of Toll-like receptor 4 (TLR4) signal transduction-related molecules expression in the tissues of the elderly. Methods: Fifty-five aged rats were randomly divided into control group, model group, toxin group and lomefloxacin group. There were 25 model rats in the model group and 10 rats in the other three groups. Endotracheal intubation, injection of Klebsiella pneumoniae, caused by multiple organ damage by pneumonia; immunohistochemical method, reverse transcription polymerase chain reaction method detection of lung, heart and small intestine TLR4 signal transduction pathway related molecular changes. Results: Compared with the control group, the levels of lipopolysaccharide-binding protein (LBP), CD14, TLR4, interleukin-1 receptor-associated kinase-1 , IRAK-1 mRNA and TLR4, TRAF6, NF-κB protein expression (P <0.01) , P <0.05). Compared with the model group, the expressions of LBP, CD14 and TLR mRNA and the protein expressions of TLR4, TRAF6 and NF-κB in lung, heart and intestine were significantly decreased (P <0.01, P <0.05). Conclusion: Toxin Qing may reduce the damage of organ tissue by decreasing the signal transduction activity of TLR4 and then reducing the secretion of cytokines. Toxin-clearing can reduce the activity of various molecules such as LBP, CD14, TLR4, IRAK-1, TRAF6 and NF-κB in the Toll-like receptor signaling pathway, which is different from that of a single inhibitor acting on only one aspect.