| Objective: To establish a COPD rat model through smoke exposure combined with lipopolysaccharide modelling,and to study the effects of Qingfei Huatan Tang on airway inflammation and PI3K/Akt/m TOR signalling pathway in COPD rats and its possible mechanism of action,so as to obtain evidence for further clinical investigation of Qingfei Huatan Tang in the treatment of COPD.Methods: Sixty male SD rats were divided into normal control group,model group,roxithromycin group,low dose group,medium dose group and high dose group according to the random number table method,10 rats in each group.The experimental animal model of COPD was established on day 1and day 14 by using a two-factor combination of intratracheal lipopolysaccharide(LPS)and fumigation,and the modeling period was 28 days.The normal control group and the model group were given 10ml/kg-d saline gavage,the roxithromycin group 30.45mg/kg-d gavage,and the low,medium and high dose groups of Qingfei Huatan Tang were given 5.69g/kg.d,11.37g/kg.d and 22.74g/kg.d gavage respectively.The drug intervention was administered once a day for 2 weeks starting on the 29 th day of moulding.After the drug intervention,the lung histopathological changes were detected by HE staining;TNF-α and IL-6 levels in serum and BALF were detected by enzyme-linked immunoassay(ELISA);PI3K,Akt and m TOR protein expressions were detected by Wester Blot.Results:1.General condition of the rats: The rats in the blank control group had normal diet,mental and activity.The rats in the model group had yellowish fur,less lustrous,slow movement,fatigue,lethargy,less food intake and light yellow secretion from the mouth and nose.Through observation,with the process of fumigation,the mental state of the rats in the model group became depressed,the fur was withered and easily shed,mucous yellow secretion was visible from the mouth and nose,the cough was further aggravated,sputum sound,shallow and rapid breathing,sometimes stridor sound,and the excretion was reduced.After the drug intervention,compared with the model group,after treatment with roxithromycin as well as low,medium and high doses of Clear Lung and Phlegm Soup drug,the rats showed improvement in cough,sputum sound and wheezing,feeding and mobility,with more obvious improvement in the roxithromycin and high dose groups.2.Histopathology of the lungs: in normal control rats,the bronchial structure was intact,the tracheal wall was not broken,the alveoli were evenly arranged,and there was no obvious inflammatory exudate: in model rats,a large number of inflammatory cells were distributed,mainly lymphocytes,the bronchial lining was thickened,the wall was thickened and irregular in shape,the lumen was narrowed,the air lumen was widened,the alveolar wall was thinned,and even the alveolar septum was destroyed,adhered and fused to form alveoli.Compared with the model group,the lung histopathology of the rats in the rosmarinic acid group and the rats in the Qingfei Huatan Tang group improved to different degrees,and the inflammatory cell infiltration in the airways was reduced,among which,the improvement was most obvious in the Qingfei Huatan Tang high-dose group.3.Expression levels of TNF-αand IL-6 contents in serum and BALF:compared with the normal group of rats,IL-6 and TNF-αcontents in serum and BALF of the model group were significantly higher(P<0.001)The difference was statistically significant,which indicates that our COPD model making can effectively stimulate the expression of inflammatory factors.The comparison of IL-6 and TNF-αin serum and BALF of each administration group with the model group revealed that the levels of IL-6 and TNF-αin serum and BALF were lower than those in the model group(P<0.05)the difference was statistically significant.Compared with the model group,the levels of IL-6 in serum and BALF decreased in the high-dose group and the western medicine group(P<0.05),among which the differences in IL-6 in BALF(P<0.01)were statistically significant,and the levels of TNF-α were significantly lower than those in the model group(P<0.001)The differences were statistically significant,indicating that the treatment could effectively inhibit the expression of inflammatory factors and was positively correlated with the dose of Chinese medicine The difference was statistically significant.The difference between the levels of IL-6 and TNF-αin the serum and BALF of the rosithromycin and high-dose groups was found to be statistically insignificant(P>0.05).4.The expression levels of PI3 K,Akt and m TOR in lung tissues were detected by Westein Blot method: compared with the blank control group,the expression of PI3 K,AKT and m TOR in the model group increased(P<0.001),and the difference was statistically significant.Compared with the model group,the expression of PI3 K,AKT and m TOR decreased in the western medicine roxithromycin group and each dose group of Chinese medicine Qing Fei Hua Tan Tang(P<0.05),among which,the decrease was most significant in the high dose group of Chinese medicine Qinglung Huayu Tang,indicating that the high dose group of Qing Fei Hua Tan Tang had an inhibitory effect on PI3 K,AKT and m TOR protein expression,and it was more obvious.Compared with the western medicine roxithromycin group,the expression of PI3 K and m TOR protein decreased in the high-dose group,but it was not statistically significant(P>0.05);the expression of AKT protein decreased,which was statistically significant(P<0.05).Conclusion:1.Qing Fei Hua Tan Tang reduces the expression of PI3 K,Akt and m TOR proteins,down-regulates the levels of IL-6 and TNF-α inflammatory factors in serum and BALF,and attenuates the inflammatory response of lung tissue in COPD rats.2.Qing Fei Hua Tan Tang clearing soup may improve airway inflammatory response in COPD rats by regulating PI3K/Akt/m TOR signaling pathway. |
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