The Expression And Effect Of IL-33,IL-8 And NETs In Pulmonary Tissues Of Patients With Chronic Obstructive Pulmonary Disease

Objective:1.To detect the expression levels of interleukin-33(IL-33),interleukin-8(IL-8)and citrullinated histone3(Cit H3)and myeloperoxidase(MPO),the main components of neutrophil extracellular trap(NETs),in lung tissues of patients with chronic obstructive pulmonary disease(COPD)and those in control group.2.To investigate the correlation between IL-33,IL-8 and NETs indicators and their correlation with clinical indicators to provide a theoretical basis for further mechanistic studies.Methods:1.A total of 77 patients who had undergone pulmonary lobectomy in the Department of Thoracic Surgery,First Affiliated Hospital of Shihezi University from October 2021 to January 2023 were included and grouped according to preoperative pulmonary function tests,including 20 cases in the non-smoking control group,18 cases in the smoking control group,19 cases in the non-smoking COPD group,and 20 cases in the smoking COPD group.2.Record general clinical information of the patient,HE staining was used to observe the histopathological morphology of lung tissue,immunohistochemical staining was used to detect the expression levels of IL-33,IL-8,Cit H3 and MPO proteins in lung tissue,immunofluorescence was used to observe the formation of NETs in lung tissue of each group,and Real-time quantitative PCR(q RTPCR)was used to detect the relative expression of IL-33 and IL-8 m RNA in lung tissue.3.Graph Pad Prism8.0 was used for mapping and SPSS 25.0 for statistical analysis.Analysis of variance was used to compare the data of normal distribution,and Bonferroni method was used to compare the data of two groups.The Kruskal-Wallis rank sum test was used to compare the data of non-normal distribution.Chi-square tests were used to compare count data sets.The correlation between genes was evaluated by Spearman correlation analysis.Two-sided test P < 0.05 was regarded as statistically significant.Results:1.Comparison of lung tissue inflammatory pathology scores between groups: The airway Bosken pathology score in the smoking COPD group was higher than that in the non-smoking COPD group,the smoking control group and the non-smoking control group(P < 0.05);the airway Bosken pathology score in the non-smoking COPD group was higher than that in the smoking control group and the non-smoking control group(P < 0.05);the airway Bosken pathology score in the smoking control group was higher than that in the non-smoking control group(P < 0.05).The airway Bosken pathology score in the smoking group was higher than that in the non-smoking group(P < 0.05).2.Comparison of degree of emphysema in lung tissue between groups: The mean lining interval(MLI)was higher in the smoking COPD group and the non-smoking COPD group compared with the smoking control group and the non-smoking control group(P < 0.05);the mean alveolar number(MAN)was lower in the smoking COPD group and the non-smoking COPD group compared with the smoking control group and the non-smoking control group(P < 0.05).3.Comparison of IL-33 expression levels between groups: the relative expression of IL-33 protein and m RNA in lung tissues of patients in the smoking COPD group was higher than that in the non-smoking COPD group,the smoking lung function normal group and the non-smoking lung function normal group(P< 0.05);the relative expression of IL-33 protein and m RNA in lung tissues of patients in the non-smoking COPD group was higher than that in the smoking lung function normal group and the non-smoking lung function normal group(P < 0.05).The relative expression of IL-33 protein and m RNA in lung tissues was higher in the non-smoking COPD group than in the smoking and non-smoking control groups(P < 0.05).4.Comparison of IL-8 expression levels between groups: The relative expression of IL-8 protein and m RNA in the lung tissue of patients in the smoking COPD group and the non-smoking COPD group were higher than those in the smoking control group and the non-smoking control group(P < 0.05).5.Comparison of Cit H3 and MPO expression levels between groups: the expression of Cit H3 and MPO proteins in lung tissue was significantly higher in the smoking COPD group and the non-smoking COPD group than those in the smoking control group and the non-smoking control group(P < 0.05).6.The results of correlation analysis showed that IL-33,IL-8 and Cit H3 were negatively correlated with FEV1/FVC,FEV1%pred and MAN(P<0.05)and positively correlated with Bosken score and MLI(P<0.05).MPO was negatively correlated with FEV1/FVC(P<0.05)and positively correlated with Bosken score(P<0.05).IL-8 positively correlated with the expression of Cit H3 and MPO(P<0.05).IL-33 positively correlated with the expression of IL-8,Cit H3 and MPO(P<0.05).Conclusion:1.The expression levels of IL-33,IL-8 and the major components of NETs,Cit H3 and MPO in the lung tissue of COPD patients were increased,and the expression of IL-33 protein was correlated with the expression of IL-8 and NETs,IL-33,IL-8 and NETs may be involved in the chronic inflammatory reaction of COPD.2.The levels of IL-33,IL-8,Cit H3 and MPO,the major components of NETs,were correlated with pulmonary function,airway inflammation score and degree of emphysema in COPD patients,it is suggested that IL-33,IL-8 and NETs may be related to the severity of COPD.