清肺化痰颗粒对慢性阻塞性肺疾病急性加重期大鼠的保护作用

The protective effects of Qingfeihuatan granules on acute exacerbation of chronic obstructive pulmonary disease in rats based on NLRP3/capase 1 pathway

目的基于NLRP3/capase1通路探讨清肺化痰颗粒对慢性阻塞性肺疾病急性加重期大鼠的保护作用。方法将SD大鼠60只随机分组为假手术组、模型组、清肺化痰颗粒(1 g/kg)组、VX-765组(NLRP3炎性小体抑制剂,剂量:50 mg/kg)、清肺化痰颗粒+VX-765组(剂量分别为1 g/kg,50 mg/kg),建立慢性阻塞性肺疾病急性加重期大鼠模型,药物处理7 d后,以全身体积描记法检测大鼠肺功能,指标包括呼吸频率(f)、潮气量(VT)、每分通气量(MV)、呼气峰流值(PEF)、吸气阻力(Ri);然后处死大鼠,采用苏木精-伊红(HE)染色检测5组大鼠肺组织病理变化,进行Holfbauer评分;采用酶联免疫吸附(ELISA)法检测5组大鼠肺泡灌洗液(BALF)中白介素-18(IL-18)、白介素-1β(IL-1β)水平,采用实时荧光定量(qRT-PCR)及免疫印迹法(Western blot)检测5组大鼠肺组织中NLRP3、caspase-1表达。结果与假手术组比较,模型组大鼠出现气管黏膜水肿充血、肺泡结构破坏,融合面积扩大、炎性细胞浸润等肺损伤症状,VT、Ri、Holfbauer评分、IL-18及IL-1β水平、NLRP3及caspase-1表达显著升高(P<0.05),f、MV、PEF显著降低(P<0.05);与模型组比较,清肺化痰颗粒组、VX-765组、清肺化痰颗粒+VX-765组大鼠病理损伤症状减轻,VT、Ri、Holfbauer评分、IL-18及IL-1β水平、NLRP3及caspase-1表达均降低(P<0.05),f、MV、PEF均升高(P<0.05);与清肺化痰颗粒组及VX-765组比较,清肺化痰颗粒+VX-765组大鼠病理损伤症状进一步减轻,VT、Ri、Holfbauer评分、IL-18及IL-1β水平、NLRP3及caspase-1表达均降低(P<0.05),f、MV、PEF均升高(P<0.05)。结论清肺化痰颗粒可以保护慢性阻塞性肺疾病急性加重期大鼠的肺组织,可能通过下调NLRP3/Capase1通路实现。

Objective To investigate the protective effects of Qingfeihuatan granules on acute exacerbation of chronic obstructive pulmonary disease(COPD)in rats based on NLRP3/capase 1 pathway.Methods The SD rats were randomly divided into sham operation group,model group,Qingfeihuatan granule(1g/kg)group,VX765 group(NLRP3 inflammatory body inhibitor,dosage:50mg/kg)group,Qingfei huatan granule+VX765 group(dosage:1g/kg,50mg/kg,respectively).The rat models with acute exacerbation of COPD were established,after 7-day drug treatment,the pulmonary function of rats was measured by whole body plethysmography,and the indexes including respiratory rate(f),tidal volume(VT),ventilation volume per minute(MV),peak expiratory flow(PEF),inspiratory resistance(Ri)were detected.Then the rats were sacrificed,and the pathological changes of lung tissues were detected by hematoxylin eosin(HE)staining,Holfbauer score was performed,the levels of IL-18 and IL-1βin alveolar lavage fluid(BALF)of rats in each group were detected by enzyme linked immunosorbent assay(ELISA),and the expression levels of NLRP3 and caspase1 in lung tissue of rats in each group were detected by real time fluorescence quantitative(qRTPCR)and Western Blot.Results As compared with the rats in sham operation group,the rats in model group showed pulmonary injury symptoms including tracheal mucosal edema,hyperemia,destruction of alveolar structure,enlargement of fusion area and infiltration of inflammatory cells,moreover the VT,Ri,Holfbauer scores,the levels of IL-18 and IL-1β,the expression of NLRP3 and caspase-1 in model group were significantly increased(P<0.05),however the levels of f,MV,PEF were significantly decreased,as compared with those in sham operation group(P<0.05).As compared with those in model group,the pathological damage symptoms in Qingfeihuatan granule group,VX765 group,Qingfei huatan granule+VX765 group were significantly alleviated,moreover the VT,Ri,Holfbauer scores,and the levels of IL-18 and IL-1β,the expressions of NLRP3 and caspase1 were significantly decreased,as compared with those in model group(P<0.05),however the levels of f,MV and PEF were significantly increased(P<0.05).As compared with those in Qingfeihuatan granule group and VX765 group,the pathological injury symptoms in Qingfei huatan granule+VX765 group were further alleviated,and the VT,Ri,Holfbauer scores,and the levels of IL-18 and IL-1β,the expressions of NLRP3 and caspase1 were significantly decreased(P<0.05),however,the levels of f,MV and PEF were significantly increased(P<0.05).Conclusion Qingfeihuatan granule can protect lung tissue of rats with acute exacerbation of COPD,and its action mechanism may be related with down-regulating NLRP3/capase-1 pathway.