Study On The Effect Of Bufei Decoction On Emphysema In COPD Mice Based On CD8+T Cell Cytotoxicit

Objective:To observe the effects of Bufei Decoction on CD8+T,granuase B(GZMB),perforin(Prf1)and Caspase3 in lung tissue of mice with chronic obstructive pulmonary disease(COPD),and study whether Bufei decoction can reduce the apoptosis of alveolar epithelial cells and improve COPD emphysema by regulating the toxicity of CD8+T cells.Methods: A total of Forty-three SPF healthy male ICR mice were randomly divided into blank control group,model group,Chinese medicine group and western medicine group,with 10 mice in the blank group and 11 mice in each group.The COPD model was established by smoking for90 days.On the 91 st day after the COPD model was established,the mice were given Bufei decoction and methylprednisolone tablets by gavage,and the relevant samples were collected after 15 days of continuous administration.The pathological changes of lung tissue were observed by HE staining,the apoptosis rate of lung tissue was observed by TUNEL,the content of CD8+T cells was analyzed by flow cytometry,and the expression of perforin and granzyme B m RNA was detected by real-time PCR.The content of Cleaved-Caspase3 in lung tissue was detected by Western-Blot.Results: 1.General status of mice in each group: the mice in the model group,the Chinese medicine group and the western medicine group showed irritability and irritability,and the respiratory rate was accelerated during smoking.After medication,the conditions of the Chinese medicine group and the western medicine group were improved.2.The pathological changes of lung tissue in each group: there was no obvious abnormality in the lung tissue of the blank group,and obvious signs of emphysema were observed in the lung tissue of the model group,and the signs of emphysema were also observed in the Chinese medicine group and the western medicine group,but they were milder than those in the model group.3.Lung function: Compared with the blank group,minute ventilation(MV),peak inspiratory flow(PIF)and peak expiratory flow(PEF)in the model group were significantly decreased.Compared with the model group,MV and PIF in the Chinese medicine group and the western medicine group were increased(P<0.01),and PEF level was not significantly different(P > 0.05).4.Flow cytometry detected the number of CD8+T cells: compared with blank group,the percentage of CD8+T cells in lung tissue of model group increased,the difference was significant(P<0.01),while the percentage of CD8+T cells in lung tissue of Chinese medicine group and western medicine group decreased(P<0.01).5.TUNEL method was used to detect lung apoptosis: compared with blank group,the lung apoptosis rate in model group was significantly increased(P<0.01),while the lung apoptosis rate in traditional Chinese medicine group and western medicine group was decreased(P<0.01).The apoptosis of alveolar epithelial cells decreased in the Chinese medicine group and western medicine group,and the apoptotic cells were mainly type II alveolar epithelial cells.6.Fluorescence quantitative PCR was used to detect the m RNA expressions of perforin and granisase B: compared with blank group,the expressions of perforin and granisase Bm RNA in lung tissues of model group were increased(P<0.01),while the expressions of perforin and granisase Bm RNA in lung tissues of traditional Chinese medicine group and Western medicine group were decreased(P<0.01).7.Cleaved-Caspase3 in lung tissue was detected by Western-Blot: Compared with blank group,the expression of lung tissue in the model group was increased,Cleaved Caspase3 was increased,and the difference was significant(P<0.01).Compared with the model group,the expression of Cleaved Caspase3 was decreased in the Chinese and western medicine groups,and the difference was significant(P<0.01).Conclusion:1.Bufei decoction can improve the pathological changes of emphysema in COPD mice,improve the lung function of COPD mice,and inhibit the apoptosis of alveolar epithelial cells.2.Bufei decoction can inhibit the cytotoxicity of CD8+T cells,suggesting that Bufei decoction may improve the degree of COPD emphysema by regulating the cytotoxicity of CD8+T cells and anti-apoptosis of alveolar epithelial cells.