HSP70与TLR-NF-κB信号途径在急性肺损伤发病机制中作用的实验研究

The investigation of the relationship between HSP70 and TLR-NF-κB during the process of acute lung injury

目的:探讨HSP70与Toll-NF-κB信号途径在急性肺损伤发病机制中的相互作用。方法:大鼠尾静脉注射脂多糖(lipopolysaccharide,LPS)制备急性肺损伤模型。将32只大鼠随机分为两组:生理盐水对照组(NS组,8只)、急性肺损伤模型组(LPS组,32只),LPS组再细分为2、6、24 h三个亚组(各8只)。LPS组尾静脉注射LPS(浓度6 mg/k g)制备急性肺损伤模型。测定各组大鼠不同时间点的动脉血气分析、肺组织湿/干重(W/D)比,ELISA法测定血清TNF-α、IL-6、IL-10,Real Time-PCR法测定肺组织HSP70、TLR及NF-κB水平并观测肺组织结构变化情况。结果:NS组大鼠各项指标无明显波动,LPS组大鼠血气分析提示存在进行性低氧、二氧化碳潴留,肺组织湿/干重(W/D)比、TNF-α、IL-6、IL-10提示逐渐升高,肺组织HSP70与TLR、NF-κB间提示随时间点推进存在正相关性(P<0.05)。结论:大鼠急性肺损伤病理过程中,HSP70与TLR、NF-κB通路间存在互相促进关系。

Objective： To investigate the possible relationship between HSP70 and Toll-NF-κB during the process of acute lung injury induced by LPS. Methods： Totally 32 adult male Wistar rats were randomly divided into two groups as follows： saline control group （NS group, 8） and lipopolysaccharide group （LPS group, 32）. Then divideLPS group into three subgroup by different time length （2/6/24 h）, each group has 8 rats. ALI was induced by injecting LPS intravenously （6 mg/kg） in the LPS group. Arterial blood gas analysis, lung tissue wet/dry weight ratio, lung histopatho.logical changes under light microscope and the scores of histological injury ineach group were observed. Serum was collected to detect the released level of TNF-α, IL-6, IL-10 by enzyme- linked immunosorbent assay （ELISA）. Real time-PCR was used to analyse the expression of HSP70, TLR andNF-κB in lung tissue. Results： No significant difference was shown in the results of NS group. As time goes on, the concentration of HSP 70 in pulmonary in LPS group was positively correlated with the concentration of TLR andNF-κB （P〈0.05）; the results of blood gas analysis suggests an progressive hypoxia and carbon dioxide retention; the W/D ratio in pulmonary raise gradually. Conclusion： During the process of rat＇s acute lung injury, there＇s aone-to-one relationship between HSP70 and Toll-NF-κB.