Objective To investigate the expression of aquaporin-1( AQP-1) in pulmonary tissues of asthma mice and the effects of acetazolamide( AZ) on AQP-1 expression. Methods Forty C57BL/6 mice were randomly divided into five groups. Group A was treated with phosphate buffer as a non-asthmatic group.The mice in group B, C, D, and E were sensitized with ovalbumin( OVA) and challenged with aerosol OVA to establish asthma model. The mice in group B, C, and D were interperitoneally injected with AZ at doses of 300, 200, 100 mg/kg, respectively during the challenge period. Results ①Wet/dry weight ratio of lung tissues in group E was significantly higher than that in group A( P lt;0. 05) , while it was lower in B, C and D groups than group E. ②The total number of cells, the number of eosinophils, and interleukin-5( IL-5) inBALF of group E were higher than those in group A( P lt;0. 05) , and interferon-γ( IFN-γ) level was lower in group E than in group A ( P lt; 0. 05) . After AZ treatment, the total number of cells, the number of eosinophils, neutrophils and lymphocytes were significantly decreased( P lt; 0. 05) , which were positively correlated with the dose of AZ. ③AQP-1 were expressed in tracheal epithelium, microvascular endothelial cell and bronchial peripheral vascular bed, and the expression in group E was significantly higher than that in group A( P lt;0. 01) . AQP-1 expression was significantly decreased after the intervention of AZ ( P lt;0. 05) .The decrease was positively correlated with the dose of AZ. The expression of AQP-1 mRNA showed no significant difference among these groups( P gt;0. 05) . Conclusions AQP-1 was over-expressed in the lung tissue of mice with asthma. AZ can inhibit the expression of AQP-1 and relieve lung inflammatory cells infiltrationin a dose-dependent manner. It is the protein expression of AQP-1 not the AQP-1 mRNA which were significantly different in different groups, suggesting that AZ affected AQP-1 in the post-transcriptional stage.
Objective To reveal the differences in gene expression levels between Th2-driven classical asthma (CA) and Th2-driven cough variant asthma (CVA) in order to investigate the pathogenesis of asthma further. Methods Clinical data were collected from asthmatic patients in the Department of Respiratory and Critical Care of Sichuan Provincial People's Hospital from June 1, 2018, to December 31, 2019. The healthy control (HC) group was healthy adults from the physical examination center. Gene expression of peripheral blood mononuclear cells (PBMCs) in the CA group, CVA group, and HC group was determined by full-length transcriptome sequencing. Differential genes were screened by GO, KEGG analysis, and protein-protein interaction (PPI) network analysis. The results of interaction network analysis were visualized by Cytoscape. Finally, the candidate genes were verified by real-time quantitative polymerase chain reaction (RT-PCR). ResultsA total of 31 patients with asthma were included in the study, including 20 patients in the CA group and 11 patients in the CVA group. According to serum total IgE > 60 IU/mL and fractional exhaled nitric oxide (FeNO) > 40 ppb as the screening condition, 9 cases of Th2-driven CA and 5 cases of Th2-driven CVA were screened for analysis. Gene expression analysis showed 300 differentially expressed genes between the Th2-driven CA group and the Th2-driven CVA group, among which 155 genes were up-regulated, and 145 were down-regulated. GO enrichment analysis showed that differential genes were mainly enriched in drug response, nitrogen compound biosynthesis, cytoplasmic matrix, protein binding, ATP binding, etc. KEGG pathway analysis showed that differential genes were mainly concentrated in 2-oxy-carboxylic acid metabolism and cytotoxic signaling pathways mediated by natural killer cells. PPI analysis revealed extensive protein interactions between different genes. Ten candidate genes were screened for RT-PCR verification and finally found that CLU, GZMB, PPBP, PRF1, PTGS1, and TMSB4X were significantly differentially expressed between the Th2-driven CA group and the Th2-driven CVA group. Conclusions Asthma's occurrence results from the interaction of many genes and pathways. CLU, GZMB, PPBP, PRF1, PTGS1, and TMSB4X genes may be essential in developing Th2-driven CVA to Th2-driven CA.
ObjectiveTo evaluate the prevalence of obstructive sleep apnea hypopnea syndrome (OSAHS) in patients with asthma, and explore the association of OSAHS with asthma.
MethodsPatients who were diagnosed as asthma between March 2014 and February 2015 were recruited in the study. They were categorized into an OSAHS group and a non-OSAHS group according to the Berlin questionnaire. The data of clinical characteristics and pulmonary function test were collected. Logistic regression analysis was performed to evaluate the factors associated with the incidence of OSAHS in asthma.
ResultsA total of 64 patients with asthma were enrolled and 36 patients were complicated with OSAHS. The body mass index (BMI), allergic rhinitis history, inspiratory capacity, maximal mid-expiratory flow and provoking dose which make FEV1 reduce 20% were significantly different between two groups (all P < 0.05). Multivariate logistic regression analysis revealed that the increased BMI was an independent risk factor of OSAHS in patients with asthma.
ConclusionThe occurrence of OSAHS with asthma is very high, and BMI may be an important associated risk factor.
Objective To investigate the effects of diesel exhaust particles ( DEP) on the production of CCL11, CCL24 and CCL26 in asthmatic rats. Methods Fifty SD rats were randomly divided into five groups. Group A was an normal control group. The rats in group B, C, D, and E were sensitized and challenged by ovalbumin ( OVA) to establish asthma model. Then the rats in the group C, D, E were inhaled DEP for 1, 2, 3 weeks, respectively. Lung tissue and brouchoalveolar lavage fluid ( BALF) were collected for detection of CCL11, CCL24, and CCL26 expression by ELISA and q-RT-PCR. Results The transcription of CCL 24, CCL26 gene and the production of CCL24 and CCL26 protein increased significantly compared with the control group ( P lt;0. 05) , and were positively associated with the DEP inhalation time. However, CCL11 gene and protein expression were not changed significantly compared with the control. Conclusion The exposure to DEP can induce the production of CCL24 and CCL26 in the asthmaic rats, which might aggravateairway hyperresponsiveness.
Objectives To observe the expression of CCL1/CCR8 mRNA in murine lung tissue of bronchial asthma and effects of glucocorticoids on their expression. Methods Thirthy healthy mice were randomly divided into a control group, an asthma group, and a dexamethasone group, with 10 mice in each group. The sensitized murine asthma model was induced by ovalbumin sensitization and challenge, and the dexamethasone group were peritoneally injected with dexamethasone( 2 mg/ kg) . Total and differential cell counts in BALF were measured. IL-4 Level in BALF was evaluated by ELISA. The expression of CCL1 and CCR8 mRNA in the lungs were measured by semi-quantitative RT-PCR. Results The percentage of eosinophils, lymphocyte and IL-4 level in the asthma group increased significantly compared with the controlgroup, and which in the dexamethasone group decreased significantly compared with the asthma group and still higher than the control group( all P lt; 0. 01) . The expression of CCL1 and CCR8 mRNA had the same tendency ( all P lt;0. 01) . Conclusions The gene expression of CCL1/CCR8 is up-regulated in allergic asthma mice.Glucocorticoids can relieve airway inflammation of asthma probably by inhibiting CCL1/CCR8 expression.
Objective To investigate the expressions of β1, 3-N-acetyl glucosaminyl transfrases ( Fringe) ( RFNG, LFNG and MFNG) in lung tissues and lung T cells isolated from asthmatic mice, and to explore the role of Fringe in pathogenesis of asthma. Methods Asthmatic BALB/ c mouse model was established by inhalation of ovalbumin after intraperitoneal injection. Meanwhile, the control groups were established by normal saline. Lung tissues were sampled after 24 hours since the last stimulation. T cells were isolated from the lung tissues using percol and NylonFiber. The mRNA expressions of three kinds of Fringe in the lung tissues and lung T cells were examined by reverse transcription-PCR ( RT-PCR) . The protein expressions of Fringe in the lung tissues were detected by Western blot. Results The mRNA expressions of RFNG, LFNG and MFNG were detectable in the lung tissues and lung T cells. The mRNA expressions of RFNG increased in the asthmatic group compared with the control group( lung tissues: 0. 92 ±0. 35 vs 0. 51 ±0. 13, P lt; 0. 01; lung T cells: 0. 33 ±0. 06 vs 0. 18 ±0. 07, P lt; 0. 01) . LFNG mRNA had lower expression level in the asthmatic group( lung tissue: 0. 77 ±0. 32 vs 1. 61 ±0. 31, P lt; 0. 01; lung T cells: 0. 49 ±0. 19 vs 0. 71 ±0. 03, P lt;0. 01) . No difference on the mRNA expression of MFNG was found in the lung tissues( 1. 44 ±0. 29 vs 1. 70 ±0. 44, P gt; 0. 05) . MFNG mRNA expression decreased in the asthmatic group compared with the control group in the T cells( 1. 17 ±0. 04 vs 0. 68 ±0. 07, P lt;0. 05) . The results of western blot were consistent with RT-PCR results of the lung tissues. The expressions of RFNG increased in the asthmatic group( 1. 17 ±0. 04 vs 0. 68 ±0. 07, P lt;0. 05) . The expression of MFNG has no difference between two groups( 8. 10 ±0. 60 vs 9. 12 ±0. 07, P gt;0. 05) . LFNG had a lower expression in the asthmatic group( 4. 11 ±0. 38 vs 6. 41 ±0. 11, P lt; 0. 05) . Conclusion The abnormal expressions of three kinds of Fringe may be involved in the pathogenesis of asthma.
