Study On The Mechanism Of Lifei Decoction In Relieving Oxidative Stress Status By MAPK/AP-1 Signaling Pathway In Chronic Obstructive Pulmonary Disease

Objective:1This study relies on rat COPD oxidative stress model,through the lung function,lung histopathological HE staining and inflammation score and oxidase markers to assess the basic efficacy of Recuperating Lung Decoction,focusing on stress levels of oxidative stress Changes in the situation,that Recuperating Lung Decoction can affect the symptoms of chronic obstructive pulmonary disease and the body's oxidative stress,the overall regulation of chronic obstructive pulmonary disease oxidation/anti-oxidative imbalance in the body state.2 MAPK/AP-1/?-GCS signaling pathway related to the kinase system was detected by molecular biology and other methods.It was revealed that the activation of the lectin system through the MAPK/AP-1/?-GCS signaling pathway COPD Oxidative Stress Realizes the Key Approach to Antioxidant.Methods:70 male Sprague-Dawley rats were randomly divided into normal group,model group,dexamethasone group,high,medium and low dose group and blockade group.Establishment of chronic obstructive pulmonary disease model.At the end of the administration,lung tissue,lung homogenate,bronchoalveolar lavage fluid and arterial blood samples were prepared from each experimental group.Pulmonary function test,pathological examination of lung injury HE staining and inflammatory pathology score.The enzyme-related immunohistochemical method was used to detect the antioxidant enzymes in the stress-related system of oxidative stress.The concentration of GST,CAT,SOD,GSH and total glutathione in bronchoalveolar lavage fluid,serum and lung tissue were measured.Immunohistochemistry,Western blot and RT-PCR were used to detect the MAPK signal pathway and its phosphorylation in lung tissue.The effect of Recuperating Lung Decoction on MAPK signal pathway was evaluated by immunohistochemistry,Western blot and RT-PCR.And y-GCS-h protein and mRNA expression,and to explore the mechanism of the effect ofRecuperating Lung Decoction on the kinase system.Results:1 experiments show that by smoking,tracheal injection of lipopolysaccharide combined with ozone exposure method,can successfully build chronic obstructive pulmonary disease rat model.After the fanning of the need for modeling group of rats that appeared bursts of cough,faster breathing symptoms,the end of the modeling sound at the end of the more obvious breathing,breathing and breathing some difficulties,throat phlegm,Apathetic,arch curled up,yellow hair and other performance.Normal group no positive symptoms,rat breathing law,good spirit,bright color shiny,quick action;blockade group of rats breathing law,the spirit can be,color luster;Recuperating Lung Decoction group and the glucocorticoid group of rats skin yellow astringent Gloss,high-dose Chinese medicine group and the glucocorticoid group to ease the symptoms.2 Recuperating Lung.Decoction can partially alleviate the lung function of chronic obstructive pulmonary disease and histopathological damage,improve airway and pulmonary inflammation.Compared with the normal group,the FEF25%and MMF of the COPD model group were lower than that of the normal group,suggesting that there was a possibility of obstruction disorder in the rats in the model group,and there may be mixed ventilation disorder Group of lung function after drug treatment after the clinical manifestations of different degrees of improvement,high-dose group and the glucocorticoid group after treatment of lung function is closer to the normal group,the middle dose group followed,but still better than the model group The blocking group was close to the normal group.Indicating that COPD rats after ventilation dysfunction and airway obstruction to get a certain degree of relief,but COPD airway remodeling performance once the formation will not change.The BALF inflammatory cell count of each group can be used to evaluate the effect of Recuperating Lung Decoction on COPD inflammation.Compared with the normal group,BALF inflammatory cells were increased in the model group.After treatment,the ratio of neutrophils and phagocytic cells was decreased in the rice and Recuperating Lung Decoction groups.The ability of the glucocorticoid group to improve the inflammation in BALF was better than that in Recuperating Lung Decoction group.HE pathology showed that Recuperating Lung Decoction group,ground glucocorticoid group and blockade group can see the bronchial lumen of the narrow,and ultimately even show occlusion,bronchial wall is thick,with varying degrees of emphysema;pulmonary inflammation Sexual cell infiltration,alveolar septum thickening,alveolar wall thinning and other pathological manifestations,but the degree of pathological damage to varying degrees of relief.Compared with the model group,the bronchial lumen of the glucocorticoid group was normal,the lumen was not thickened obviously,the inflammation of the tube wall was mild,the alveolar septum was slightly widened,the lobular center type and whole lobular emphysema were mildly formed.A large number of hormones often lead to some rats have obvious diffuse interstitial non-inflammatory performance.The dose of Recuperating Lung Decoction was higher than that of Recuperating Lung Decoction.The airway inflammation in the high dose group was mild and the stenosis was not obvious.The bronchial and vascular smooth muscle showed mild hyperplasia.Tissue injury and pathology of inflammatory pathology,we can see Recuperating Lung Decoction group showed a dose-effect relationship between the groups,the specific score of lung soup,low-dose group score was the same or slightly lower than the model group,and traditional Chinese medicine high-dose group with The glucocorticoid group is similar.And lung tissue histopathological HE staining results are basically the same.3 Recuperating Lung Decoction can effectively enhance the body's antioxidant enzyme expression,ease the body's peroxide.The ratio of GSH/GSSG in model group decreased,indicating that there was oxidative stress in the body.After treatment,the glucocorticoid group and the high and middle dose groups of Chinese medicine can improve the oxidative stress injury state in different degrees.Meanwhile,the drugs can effectively enhance the expression of antioxidant enzymes in tissues and serum.The ratio of GSH/GSSG can reflect the oxidative stress during the pathological changes of COPD.The results showed that the ratio of GSH/GSSG increased the activity of oxidase and inhibited the oxidative stress.4 MAPK/AP-1/?-GCS signaling pathway is a key link in the development of COPD.In order to explore the mechanism of Recuperating Lung Decoction in regulating the mechanism of stress kinase system,the MAPK signal pathway,the c-Jun and c-Fos nucleoprotein and mRNA and ?-GCS-h contents of AP-1 were detected by different methods.The results showed that the phosphorylation of P38 MAPK,JNK and ERK in COPD was significantly increased.After treatment with Recuperating Lung Decoction,the phosphorylation of P38 MAPK,ERK and JNK was effectively inhibited.The results showed that the stimulation of reactive oxygen species significantly increased the phosphorylation level of MAPK signaling pathway.Recuperating Lung Decoction can be reduced after treatment p38MAPK,ERK,JNK activation.The levels of c-Jun and c-Fos nucleoprotein and mRNA in the lung tissue of the high dose group were not significantly different from those in the glucocorticoid group.It is suggested that the expression of AP-1 protein and mRNA can down-regulate the oxidative stress of COPD and achieve a certain therapeutic effect through the effect of phosphorylation of MAPK signal pathway.The content of y-GCS-h nucleoprotein in the lung tissue of the high dose group was not significantly different from that of the glucocorticoid group,indicating that the Recuperating Lung Decoction could relieve the oxidative stress state of the body and the low expression of y-GCS-h protein and mRNA,So that the body to maintain a balance of oxidation and oxidation.Conclusion:1 This experiment was successfully established in rats with chronic obstructive pulmonary disease model.2 PD98059 can block MAPK-ERK1/2 signaling pathway in lung tissue of COPD rats.3 MAPK/AP-1/?-GCS is the key link of oxidative stress on the development and progression of COPD4 Recuperating Lung Decoction is effective in the treatment of COPD rats,and its mechanism involves the regulation of the kinase system.5 Recuperating Lung Decoction can inhibit the phosphorylation of MAPK signal pathway,down-regulate the expression of AP-1 protein and mRNA,and thus affect the expression of?-GCS protein and mRNA,to achieve therapeutic effect.However,the signal transduction of MAPK/AP-1/?-GCS is a very fine and complicated process.The target of Recuperating Lung Decoction is not very clear,and the improvement of the therapeutic mechanism still needs more in-depth study.