【摘要】 目的观察腹腔感染脓毒症时肺组织内病原菌相关模式分子(内毒素、脂蛋白、DNA)的主要模式识别受体的表达变化及其意义。方法采用盲肠结扎穿孔(CLP)术建立30只昆明种小鼠腹腔感染脓毒症模型,将模型小鼠随机分为CLP组和假手术组,每组各15只鼠,分别于术后8,12和24h取5只鼠处死取右肺组织,采用逆转录聚合酶链反应(RT PCR)法检测肺组织内毒素受体[清道夫受体(SR)、白细胞分化抗原14(CD14)、TLR4]及细菌脂蛋白受体(TLR2)、细菌DNA受体(TLR9)mRNA的表达,酶联免疫吸附实验(ELISA)法检测肺组织内肿瘤坏死因子α(TNFα)含量,采用分光光度计法测定肺组织髓过氧化物酶(MPO)活性。结果与假手术组比较,术后8hCLP组肺组织内CD14mRNA(1.143±0.139,t=0.022,P<0.05),TLR2mRNA(0.418±0.102,t=0.021,P<0.05),TLR4mRNA(0.595±0.052,t=0.0001,P<0.01)和TLR9mRNA(0.743±0.178,t=0.0023,P<0.01)表达上调,其中TLR9mRNA呈持续上调变化,SRmRNA(0.659±0.159,t=0.029,P<0.05)呈持续下调改变;肺组织CD14、TLR4、SR、TLR2和TLR9mRNA的表达变化分别与MPO和TNFα变化呈明显的相关关系(P<0.05或P<0.01)。结论脓毒症发病过程中,肺组织内主要模式识别受体表达变化与肺损伤相关,除LPS外,其他细菌成分也可能参与了肺损伤过程。更多还原

【Abstract】 Objective In order to investigate the evidence of the synergistic effects of bacterial components, to observe the relationship of the expression of lipopolysaccharide (LPS) receptors [CD14, Toll-like receptor 4 (TLR4), scavenger receptor(SR)], lipoprotein receptor (TLR2) and bacterial DNA receptor (TLR9) with pulmonary injury in abdominal infection-induced sepsis.Methods 30 mice were used and randomly divided into cecal ligation puncture (CLP) (n=15) and sham (n=15) groups.The animals were respectively sacrificed 8, 12 and 24 (each point n=5 ) hours following CLP and sham CLP. The lungs were removed and immediately stored in liquid nitrogen for TLRs mRNA , tumor necrosis factor(TNF) α and myeloperoxidase (MPO) assay. To detect the expression of CD14, TLR4,SR, TLR2 and TLR9 mRNA by reverse-transcription polymerase chain reaction, to detect the TNF-α content of the lung tissue by enzyme-labeled immunosorbent assay, and to assay the MPO activity of the lung tissue spectrophotometer.Results It was found that the expression of receptors for LPS, BLP and bacterial DNA in pulmonary tissues was markedly changed in CLP-induced sepsis, showing upregulation of CD14 mRNA(1.143±0.139,t=0.022,P<0.05),TLR2 mRNA(0.418±0.102,t=0.021,P<0.05)、TLR4 mRNA(0.595±0.052,t=0.0001,P<0.01)and TLR9 mRNA(0.743±0.178,t=0.0023,P<0.01)at different degrees (P<0.05 or P<0.01)after postinjury 8 h, among which the expression of TLR9 mRNA kept increasing. The expression of SR mRNA (8 h:0.659±0.159;12 h: 0.429±0.061;24 h: 0.300±0.045;t=0.029,P<0.05;t=0.001,P<0.01;t=0.003,P<0.01)showed continuous down- regulation.Conclusion There was a marked correlation between the changes of pattern-recognition receptor expression and the increases of MPO and TNF-α levels in pulmonary tissues.更多还原