Effects Of Edaravone On Glucocorticoid Resistance In COPD Rats By Anti-oxidative Stress

Glucocorticoids are widely used in the treatment of chronic inflammation,autoimmune diseases and leukemia,etc.,to play its anti-inflammatory,anti-immune effects,but not for all patients are effective.In clinical practice,glucocorticoids play an important role in the treatment of chronic obstructive pulmonary disease(COPD)and asthma.Some people are insensitive or resistant to hormones,are not conducive to the treatment of diseases,and are difficult to control the progression of the disease.There is glucocorticoid resistance.Studies have confirmed that COPD and bronchial asthma are persistent chronic airway inflammation,but the mechanism of airway inflammation are very different,for example,in airway inflammatory cells,COPD airway inflammation is mainly mainly neutrophils,While asthma is mostly eosinophils mainly.Neutrophils are key effector cells in the pathogenesis of COPD.They are significantly increased in the lungs of the COPD,associated with the severity of the disease,participating in airflow limitation,chronic bronchitis,and emphysema in the progression of the disease.Neutrophils in patients with COPD spontaneously release large amounts of reactive oxygen species(ROS),which release proteases and ROS in order to migrate through complex tissues within the body when neutrophils migrate through the lung tissue.In addition,migration may lead to more range of damage while stimulating their degranulation,including degrading cytokines such as IL-8,TNF-α,ROS and proteases(e.g.,neutrophil elastase,COPD patients increased sputum,airway structure damage),increased lung tissue in a wider area of inflammatory response and protease activation.In addition,COPD pathogenesis of oxidative stress can lead to increased ROS,the body too much,ROS in addition to a direct result of lung injury,but also can activate the inflammatory cell release of inflammatory factors,leading to neutrophil activation and aggregation,both Promote each other,thus a vicious circle,increase the broken lung tissue.Studies have shown that oxidative stress can lead to the formation of nitrite peroxides in the body,the nitration of HDAC2 tyrosine residues,leading to its inactivation,ubiquitination and degradation;In addition,oxidative stress products can activate PI3Kβ /(AKT)signaling pathway leading to phosphorylation and inactivation of HDAC2,regardless of that caused by decreased HDAC2 activity,can activate NF-k B,trigger inflammation switch,IL-17,IL-17 stimulates airway epithelial cells to produce IL-8,causing neutrophil recruitment,continue to cause the release of inflammatory factors,such as TNF-α,inflammatory cell activation,causing NF-k B Activation,which is a positive feedback regulation,neutrophils accumulate in the airway and lung tissue in large quantities,leading to glucocorticoid resistance.The above indicates that oxidative stress-induced neutrophil recruitment in airways may be an important mechanism of glucocorticoid resistance and continues to increase glucocorticoid resistance in the development of COPD disease.Edaravone is a new type of strong oxidants used in clinical practice,with a strong antioxidant effect,the current clinical application of acute cerebral infarction is also used in paraquat poisoning,acute lung injury and other treatment.Animal experiments found that edaravone can improve microcirculation,protect the cells from oxidative imbalance and inhibit the release of lysosomes and other effects.This study was designed to discuss the effects of edaravone on oxidative stress in COPD rats and to improve the effect of glucocorticoid resistance due to neutrophil recruitment in the lungs by anti-oxidative stress and its mechanism.Edaravone can inhibit the inflammatory response to the above-mentioned antioxidant activity,regulate neutrophil recruitment,thereby improving hormone resistance,there is no similar research at home and abroad.Materials and methodsHealthy male SD rats 50,weighing 200 ± 20 g.They were randomly divided into healthy blank control group(n = 10),model group(n = 40).Model group: COPD rat model was established by somking the method with dropping LPS into the airway.The model was developed for 60 days.Every 14 days,the rats were exposed to LPS once every 200 μl,and the other time was smoked in a self-made closed carton(50cm * 40 cm * 30cm).After the model was completed,the model group was randomly divided into COPD group,edaravone group,budesonide group and combined intervention group.COPD group was treated with saline intraperitoneal injection,1 time / day,2ml / only,a total of 14 days.Edaravone group was treated with intraperitoneal injection of edaravone,1 time / day,0.2mg / 100 g,a total of 14 days.Budesonide group was treated with aerosolized budesonide,1 times / day,1mg / only,a total of 14 days.Joint intervention group: given edaravone intraperitoneal injection combined with budesonide inhalation,dose and time ibid.The serum levels of IL-8,IL-17,TNF-α,and NF-k B in serum and alveolar lavage fluid were measured by enzyme-linked immunosorbent assay(ELISA).The pathological changes of lung tissue were observed after being staining HE.Statistical analysis: All data were analyzed using SPSS21.0 statistical software.The data were described as mean ± standard deviation((?)±s).The t test was used between the two groups.Single factor analysis of variance(ANOVA)was used for comparison among groups.Pearson correlation was used for correlation analysis.Significant test level was α=0.05.Results1 the pathological changes of lung tissues in ratsCompared with the blank group,the basic structure of the lung tissue in the COPD group was normal,the tracheal and bronchial mucosal epithelial cells were swollen,disorderedand decreased.The small airway showed mucus plug and a lot of inflammatory cell infiltration,bronchial smooth muscle thickening,alveolar wall Thinning,rupture,alveolar enlargement,multiple alveolar lumens,mutual melting and even rupture into the lungs.This is consistent with the histopathological changes of trachea and lung tissue in COPD patients.The degree of lung injury in rats was lighter than that in COPD group,and the lung tissue destruction in the intervention group was relatively light.2 Changes of pulmonary function in each group ratsThe levels of FEV0.3 / FVC and PEF were lower in COPD group than those in control group(P<0.05).The levels of FEV0.3 / FVC and PEF in the combined intervention group were significantly higher than those in the COPD group,the budesonide group and the edaravone group,and the difference was statistically significant(P<0.05).3 Leukocyte and neutrophil levels in arterial and alveolar lavage fluid of ratsThe percentages of white blood cells and neutrophils in the arterial and alveolar lavage fluid of the COPD group were significantly higher than those of the blank group(P<0.05).The white blood cell and neutrophil counts of arterial blood and alveolar lavage fluid in edaravone group were significantly lower than those in COPD group(P<0.05),but the difference was not statistically significant(P<0.05).The percentage of white blood cells and neutrophils in the arterial blood and alveolar lavage fluid of the intervention group were significantly lower than those of the COPD group and budesonide and edaravone group,and the difference was statistically significant(P< 0.05).4MDA and SOD levels in each serumThe level of MDA in budesonide group was significantly lower than that in COPD group(P<0.05),and the level of MDA in COPD group was significantly higher than that in blank group,SOD level was lower than that in blank group,and the difference was statistically significant(P<0.05).The level of serum MDA in edaravone group was significantly lower than that in COPD group(P<0.05),and the difference was statistically significant(P< 0.05).The levels of serum MDA and SOD in the combined intervention group were significantly lower than those in the COPD group and budesonide group and the edaravone group(P<0.05).5IL-8,IL-17,TNF-α,NF-k B levels in each serum and alveolar lavage fluidThe levels of IL-8,IL-17,TNF-α and NF-k B in the COPD group were obviously higher than those in the blank group,and the differences were statistical significance.(P <0.05).The levels of IL-8,IL-17,TNF-α and NF-k B in the budesonide group and the edaravone group were significantly lower than those in the COPD group,but the difference was not statistically significant(P> 0.05).The levels of IL-8,IL-17,TNF-α and NF-B in the combined intervention group were significantly lower than those in the other three groups,but the difference was not were statistical significance(P> 0.05)6 Serum and alveolar lavage fluid(F)IL-17,IL-8,NF-k B correlation analysisIL-17,IL-8 and TN-k B were positively correlated with inflammatory factors in alveolar lavage fluid,r = 0.942(P <0.05),r = 0.912(P <0.05),r(P <0.05).The levels of IL-17 in serum were positively correlated with IL-8 and NF-k B in alveolar lavage fluid,r = 0.725(P <0.05),r = 0.776(P <0.05)(P <0.05),r = 0.801(P <0.05).The serum levels of NF-κB and NF-κB were significantly correlated with IL-17 and NF-k B in alveolar lavage fluid,r =0.724(P =0.05),r = 0.768(P <0.05).IL-17 and IL-8 are positively correlated with alveolar lavage fluid.7 Correlation between IL-17,IL-8,NF-k B and serum MDA and SOD in serum(B)and alveolar lavage fluid(F)The levels of IL-17 and MDA in serum and alveolar lavage fluid were positively correlated with rB = 0.642(P<0.05),rF =0.675(P<0.05).The levels of TN-k B in serum and alveolar lavage fluid were positively correlated with MDA level,rB = 0.365(P<0.05),rF = 0.387(P<0.05).The levels of IL-17 in serum and alveolar lavage fluid were negatively correlated with SOD level,rB =-0.425(P<0.05),rF =-0.457(P<0.05);rat serum and alveolar lavage fluid IL-8 and SOD levels were negatively correlated,respectively rB =-0.645(P<0.05),rF =-0.685(P<0.05).The levels of IL-8 were negatively correlated with SOD level in theserum and alveolar lavage fluid,rB =-0.376(P<0.05),rF =-0.396(P<0.05).Conclusion1.The levels of IL-8,NF-k B and neutrophils in the blood and alveolar lavage fluid of COPD rats were higher than those in the COPD rats.The levels of IL-8,NF-k B and neutrophil were increased,the level of SOD was decreased,the level of MDA was increased,/ FVC decreased,COPD rats were with airflow limitation.2.The levels of IL-17 and neutrophils in the blood and alveolar lavage fluid in the budesonide group were lower than those in COPD rats,but there was no statistically significant resistance to glucocorticoid induced by neutrophil recruitment.3.Edaravone can reduce oxidative stress in COPD rats.4.Edaravone combined with budesonide in the activation of NF-k B by anti-oxidative stress,could reduce IL-17 and neutrophil levels and improve glucocorticoid resistance in rats.