The Role Of SERPINB10 Gene In Airway Eosinophilic Inflammation Of Asthma

Part?The expression of SERPINB10 in airway epithelium of asthmatic patients and its association with airway eosinophilic inflammation in asthmaObjective:To investigate the expression of SERPINB10 in mRNA and protein levels in endobronchial brushing samples and airway biopsies,to explore the position of SERPINB10 and to clarify the relationship between SERPINB10 levels and airway eosinophlic inflammation in subjects with asthma.Methods:We recruited 13 healthy control subjects and 40 subjects with asthma.All subjects were recruited from Tongji Hospital.Subjects with asthma were diagnosed by a physician;had symptoms of episodic cough,wheeze and/or dyspnea;had accumulated dosage of methacholine provoking a 20%fall of forced expiratory volume in the first second(FEV₁ PD₂₀)<2.505mg and/or?12%increase in FEV₁following inhalation of 200ug salbutamol.Control subjects had no respiratory symptoms,normal spirometric values and methacholine PD₂₀>2.5mg.None of the subjects had ever smoked or received inhaled or oral corticosteroid.For each subject,we recorded demographic information,collected induced sputum samples,measured spirometry and fraction of exhaled nitric oxide?FeNO?.We performed bronchoscopy with brushing and endobronchial biopsy.We examined the expression of SERPINB10gene in endobronchial brushings by real-time PCR and observed the position of SERPINB10 in airway biopsies by immunohistochemistry.To investigate the association between SERPINB10 levels and eosinophilic inflammation,we analysed the correlation between the expression of SERPINB10 and the percentage of sputum eosinophilia,biopsy eosinophils and FeNO in subjects with asthma.Results:We found that subjects with asthma had lower FEV1 and methacholine PD₂₀than the health control subjects.We found that the Serpinb10 transcript levels were markedly increased in subjects with asthma when compared with healthy controls?P<0.0001?.Serpinb10 immunostaining in airway biopsy samples revealed that Serpinb10-positive cells were mainly bronchial epithelial cells and the majority of Serpinb10 was located in the nucleus.Correlation analysis suggested that Serpinb10transcript levels were significantly correlated with the percentage of sputum eosinophils?r=0.3710,P=0.0184?and the number of eosinophils in bronchial submucosa?r=0.4572,P=0.0030?in subjects with asthma.There was markedly correlation between Serpinb10 transcript levels and FeNO?r=0.3446,P=0.0295?.Conclusion:SERPINB10 in asthmatic patients was mainly located in the nucleus of airway epithelial cells.Compared with healthy controls,the expression of SERPINB10 in airway brushing samples of subjects with asthma showed a significantly increase in mRNA and protein levels.The SERPINB10 transcript levels in airway epithelial cells of asthmatic patients were positively correlated with airway eosinophilic inflammation.Part ? Mechanism of SERPINB10 Gene Involved in Airway Eosinophilic Inflammation of Asthma Objective: To explore the specific mechanism of SERPINB10 gene involvement in airway inflammation of asthma by using asthmatic cell model.To investigate the role of SERPINB10 in IL-13-induced expression of inflammatory cytokines in human bronchial epithelial cells.To analysis the correlation between expressions level of SERPINB10 and inflammatory cytokines in airway brushings in patients with asthma.Methods: We used BEAS-2B cell line and primary normal human bronchial epithelial?NHBE?cells in our study and used ordinary culture and air-liquid interface culture,respectively.The expression of SERPINB10 in cells was inhibited by transfection of SERPINB10 si RNA.Cells were divided into four groups:?A?control group?B?IL-13 intervention group?C?transfection group and?D?intervention after transfection group.The expression of SERPINB10,POSTN and CCL26 in m RNA level was detected by real-time PCR.The expression of POSTN protein in the culture supernatant was detected by ELISA.The expression of POSTN in airway brushings was detected by Real-time PCR and the correlation between the expression of POSTN and the expression level of SERPINB10 in airway brushings was nanlyzed.Results: Compared with control group,the expression of SERPINB10,POSTN and CCL26 in IL-13 intervention group was increased remarkably.Similar trend appeared in intervention after transfection group when compared with transfection group.While compared with intervention group,the expression of SERPINB10,POSTN and CCL26 in intervention after transfection group was significantly decreased and differences were statistically significant.The expression level of SERPINB10 was significantly positively correlated with the expression level of POSTN in airway brushings in patients with asthma.Conclusion: The expression of SERPINB10 in BEAS-2B and NHBE cells was inhibited after transfection with human SERPINB10 si RNA and the expression of inflammatory cytokines POSTN and CCL26 was also decreased.It was concluded that SERPINB10 gene plays an important role in the expression of IL-13-induced inflammatory cytokines POSTN and CCL26 in airway epithelial cells.