Based On "CS Exposure-induced T Cell Immune Response" To Explore The Mechanism Of COPD Airway Mucus Hypersecretion And The Effect Of Jinshui Liujun Decoction

Objective:To elucidate the possible pathological mechanism of T cell autoimmune response in a mouse model of airway mucus hypersecretion in chronic obstructive pulmonary disease(COPD)and the mechanism of action of Jinshui Liujun decoction.Methods:Thirty two male ICR mice were randomly divided into a blank group,a model group,a Jinshui Liujun decoction group and a methylprednisolone group,with 8 mice in each group.Except the normal group,the rest of the groups induced COPD models by smoking for 43 days and instilling mouse Elastase into the trachea for a total of 43 days.They rested for 2 weeks before instilling Elastin into the trachea,and then instilled Elastin into the trachea for three consecutive days,with a concentration of 2mg/ml,100ug/0.1ml elastin and 0.1ml Freund’s adjuvant were intraperitoneally injected into rats.On the 61st day,Jinshui Liujun decoction was administered by gavage.After 14 days of gavage,lung function was tested under anesthesia and specimens were collected.HE staining method was used to observe the pathological changes of mouse lung tissue;PAS method was used to detect the distribution of airway goblet cells;Flow cytometry was used to detect the expression of CD4 and CD8 in lung tissue;ELISA was used to detect the contents of matrix metalloproteinase-12(MMP-12)and interleukin-17a(IL-17a)in alveolar lavage fluid;Western blot method was used to detect the expression of MUC5AC.Results:1)Evaluation after HE staining:the trachea,lung parenchyma,and pulmonary blood vessels of the blank group mice showed no significant abnormalities,and the size of the alveoli was basically normal,with regular shape,without inflammatory cell infiltration;In the model group and the Jinshui Liujun Decoction group,the intrapulmonary alveolar ducts,alveolar sacs,alveoli,and bronchioles were significantly enlarged compared to the blank group,with alveolar walls becoming thinner and having multiple fractures,and adjacent alveoli fusing with each other to form a larger lung mass.The alveolar septum was destroyed,part of the alveoli fused into the pulmonary bullae,bronchial epithelial mucosa was partially detached,and phlegm and mucus retention could be seen in the lumen,but the above lesions in the Jinshui Liujun Decoction group were lighter than those in the model group.2)In the blank group,a small amount of scattered goblet cells can be seen in the bronchial mucosa of mice,while in the model group,the number of goblet cells in the trachea and bronchi of mice significantly increased,and the secretion of mucin in the goblet cells was increased,and a small amount of PAS positive secretion was visible in the lumen.Compared with the blank group,the number of airway goblet cells and the amount of mucus in the model group were significantly increased(P<0.01);Compared with the model group,the number of goblet cells and the amount of mucus in the Jinshui Liujun Decoction group were significantly reduced(P<0.05,P<0.01).3)Compared with the blank group,the expression of MUC5AC protein in the lung tissue of mice in the model group,Jinshui Liujun Decoction group,and methylprednisolone group significantly increased(P<0.01).Compared with the model group,the expression of MUC5AC protein in the lung tissue of mice in the Jinshui Liujun Decoction group and methylprednisolone group significantly decreased(P<0.01).4)Compared with the blank group,the expression of CD4 in the lung tissue of mice in the model group,the Jinshui Liujun Decoction group,and the methylprednisolone group decreased at different levels,while the expression of CD8 increased,while the expression of CD4/CD8 decreased(P<0.05),with statistical significance.Compared with the model group,the expression of CD4 in the lung tissue of mice in the Jinshui Liujun Decoction group and methylprednisolone group increased,the expression of CD8 decreased,and the expression of CD4/CD8 increased(P<0.05),with statistical significance.5)Compared with the blank group,the expression of MMP-12 and IL-17a protein in BALF of mice in the model group,Jinshui Liujun Decoction group,and methylprednisolone group was significantly increased(P<0.01),with statistical significance;Compared with the model group,the expression of MMP-12 protein in BALF of mice in the Jinshui Liujun Decoction group and methylprednisolone group was significantly decreased(P<0.01),and the expression of IL-17a protein was significantly decreased(P<0.01),with statistical significance.Conclusion:Jinshui Liujun Decoction can improve airway mucus hypersecretion in COPD mice by regulating T cell immune response induced by CS exposure.Jinshui Liujun Decoction can significantly downregulate the expression of MMP-12 and IL-17a proteins,increase the expression of CD4 cells,reduce the expression of CD8 cells,enhance humoral and cellular immunity,significantly reduce the expression of MUC5AC protein in lung tissue,and improve airway mucus hypersecretion in COPD mice.