通塞颗粒对AECOPD痰热证大鼠肺组织TLR4信号通路的影响

Influence of Tongsai Granule on toll-like receptor 4 signal transduction pathway in lung tissue of rat models with acute exacerbation of chronic obstructive pulmonary disease accompanied by phlegm-heat syndrome

目的观察慢性阻塞性肺疾病急性加重期(AECOPD)痰热证大鼠肺组织TLR4信号通路,探讨通塞颗粒治疗AECOPD痰热证的作用机制。方法制备AECOPD和AECOPD痰热证模型,将50只Wistar大鼠分为空白对照组、AECOPD组、AECOPD痰热证组、阳性对照组及通塞颗粒组。肺组织TLR4、NF-κB、IL-1β、TNF-α、IL-6蛋白表达采用免疫组化法测定,肺组织IL-1βmRNA的表达采用RT-PCR法测定。结果与空白对照组相比,AECOPD组、AECOPD痰热证组肺组织TLR4、NF-κB及其下游炎症因子(IL-1β、TNF-α、IL-6)表达均显著增强(P<0.05,P<0.01),其中,AECOPD痰热证组上述指标表达水平较AECOPD组增强明显(均P<0.05);与AECOPD痰热证组比较,阳性对照组、通塞颗粒组上述指标表达均显著减弱(P<0.05,P<0.01),其中,通塞颗粒组IL-6较阳性对照组降低更明显(均P<0.05)。结论TLR4通路活化参与了AECOPD痰热证的发生发展。通塞颗粒治疗AECOPD痰热证的机制可能与其抑制TLR4信号转导通路,从而减轻该通路引发的炎症损伤有关。

Objective To observe the toll- like receptor 4 signal transduction pathway in the lung tissues of rat models with AECOPD accompanied by phlegm- heat syndrome and to approach the mechanism of Tongsai Granule,in treatment of the disease.Methods 50 Wistar rats were randomly divided into five groups： normal,AECOPD,phlegm- heat syndrome of AECOPD,positive control and Tongsai Granule treated groups（ each,n = 10）. TLR4,NF- κB,IL- 1β,TNF- α and IL- 6 protein expressions in the rat lung tissues were analyzed by immunohistochemistry staining. IL- 1β mRNA expression level in the rat lung tissues was measured by reverse transcription- polymerase chain reaction（ RT- PCR）. Results TLR4,NF- κB,IL- 1β,TNF-α and IL- 6 proteins and IL- 1β mRNA expression levels in AECOPD group and phlegm- heat syndrome of AECOPD group were markedly higher than those in normal group（ P 0. 05,P 0. 01）,and in phlegm- heat syndrome of AECOPD group the expressions were significantly higher than those in the AECOPD group（ all P 0. 05）. Compared with phlegm- heat syndrome of AECOPD group,the lung tissue expressions of TLR4,NF- κB,IL- 1β,TNF- α and IL- 6 proteins and IL- 1β mRNA were markedly weakened in positive control group and Tongsai Granule treated group（ P 0. 05,P 0. 01）. The expressions of IL-6 protein in Tongsai Granule treated group were significantly lower than those in the positive control group（ P 0. 05）. Conclusion TLR4 signal transduction pathway was involved in the pathological changes of lung tissues in phlegm- heat syndrome of AECOPD. The mechanism of Tongsai Granule in treating phlegm- heat syndrome of AECOPD may be related to its ability of suppressing TLR4 signal transduction pathway.