| Objective: To observe the effect of Qingjin Huatan Granule on chronic obstructive pulmonary disease(COPD)acute exacerbation(phlegm heat obstructing the lung)in the lung tissues of rats with JAK/STAT signal transduction pathway in the activation of transcription factor 1,3(STAT1,STAT3)on the expression of Qingjin Huatan with high dose,and then to explore the mechanism of Qing Jin Huatan Granule in preventing and treating acute exacerbation of chronic obstructive pulmonary disease,so as to provide experimental evidence for its clinical application and a reference for Chinese medicine in preventing and treating chronic obstructive pulmonary disease.Methods: 40 healthy male SPF grade SD rats with weight(200 + 50)g were used.All rats divide into 5 groups of 8 in each stochastically,blank group、model group、group of Qing Jin Huatan Granule with low dose,group of Qing Jin Huatan Granule with high dose and western medicine group.The smoke exposure +LPS(lipopolysaccharide)combined with the classic preparation method of modeling,using Qingjin Huatan Granule as treatment group was divided with two kinds of high and low dose.Roxithromycin is the western medicine control group,moreover,the normal control group and the model group were set up,samples received drug treatment.The low and high dose of Qing Jin Huatan Granules were fed at 9.4,37.6g/kg-1g respectively.Roxithromycin group was treated by 0.0315g·kg-1g fed rats,normal controlgroup and model group with the same volume of saline.The drug was given once a day for two weeks.The total number of white blood cells(×109)and the percentage(%)of neutrophils in the peripheral blood of rats were measured.HE staining was used to observe the histological changes of lung.Meanwhile,ELISA was used to detect the contents of IL-1 beta and TNF-alpha cytokines.The expression levels of IL-1 beta、TNF-alpha,STAT1 and STAT3 mRNA in lung tissue were detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR).The expression of STAT1 and STAT3 protein in lung tissue was detected by Western blot.Results: 1.Pathological sections of lung tissueHE staining of lung tissue in the model group show that inflammatory cell infiltration around the bronchus,airway wall thickening,lumen occlusion,stenosis,the alveolar structure was destroyed,The alveoli were dilated and fused into a pulmonary bullae,and exudate was seen in the alveoli,suggesting that the model was successful.The extent of damage of Roxithromycin group and Qingjin Huatan Granule with high dose on the lung tissue were improved.2.The percentage of white blood cells and neutrophils in the peripheral blood of rats was reduced by Qingjin Huatan Granule.Low dose group was lower than Roxithromycin group and Qingjin Huatan Granule with high dose group(P<0.05)(considerable efficacy(P> 0.05)).3.Changes in the content of IL-1 beta and TNF-alpha in BALF of rats in each groupCompared with the blank group,the content of IL-1 beta,TNF-alpha BALF in model group were significantly increased(P<0.01);Roxithromycin group and Qingjin Huatan Granule with high dose IL-1 beta,TNF-alpha was significantly decreased,no significant difference between the two(P > 0.05);Qingjin Huatan Granule with low dose group BALF beta,IL-1 TNF-alpha decreased levels below the Roxithromycin group(P<0.05).4.Changes of mRNA expression of IL-1 beta,TNF-a,STAT1 and STAT3 in lung tissue of ratsCompared with the normal group,the expression of IL-1 beta,TNF-alpha,STAT1 and STAT3 mRNA were significantly increased(P<0.01)in the lung tissue model group;Compared with the model group,Roxithromycin group,Qingjin Huatan Granule with low and high dose groups can significantly reduce the lung tissue of rats with IL-1,TNF-,STAT1 and alpha beta STAT3expression(mRNA P<0.01).Roxithromycin group and Qingjin Huatan with high dose group decreased more significantly,no obvious difference between them.5.Expression level of STAT1 and STAT3 protein in lung tissue of ratsThe expression of STAT1 and STAT3 protein in lung tissue of rats in each group compared with the normal group.STAT1 and STAT3 in lung tissue of model rats increased significantly(P<0.01);Compared Roxithromycin group,Qingjin Huatan Granule groups with model group,STAT1 and STAT3 in lung tissue of rats was significantly reduced(P<0.01).Roxithromycin group and Qingjin Huatan Granule with high dose group decreased more significantly,there was no significant difference between Roxithromycin group and Qingjin Huatan Granule with high dose group.Conclusion(s): Qingjin Huatan Granule can confer protection and relieve the airway inflammation to the lung tissue.The mechanism may inhibit the increase of the level of IL-1 beta and TNF-alpha by declining the expression and activation of STAT1 and STAT3 in JAK/STAT signaling pathway.Thus the inflammatory response can be reduced and the acute attack and aggravation of COPD. |
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