Objective To observe the eotaxin expression of rat airway smooth muscle cells ( ASMCs) induced by serum from asthmatic rats, and explore the possible mechanism. Methods ASMCs isolated fromrat tracheas were cultured in vivo. Then they were treated with serum from asthmatic rats, or treated with serum and dexamethasone simultaneously. The level of eotaxin protein in supernatant and eotaxin mRNA in ASMCs were measured by ELISA and reverse transcription-polymerase chain reaction. The expression of cAMP in ASMCs was examined by radioimmunoassay. Results After the treatment with sensitized serum, the eotaxin level in supernatant and mRNA expression in ASMCs were significantly higher [ ( 107. 09 ±7. 12) ng/L vs. ( 0. 63 ±0. 56) ng/L, P lt; 0. 05; 1. 39 ±0. 04 vs. 0. 05 ±0. 01, P lt;0. 05] , and the level of cAMP in ASMCs was significantly lower compared with the control group [ ( 17. 58 ±3. 62) ng/L vs. ( 32. 39 ±3. 36) ng/L, P lt; 0. 05] . After intervened by the sensitized serum and dexamethasone simultaneously, the protein and mRNA expressions of eotaxin were lower compared with those intervened by sensitized serumalone [ ( 64. 18 ±4. 04) ng/L and 0. 77 ±0. 19] . The level of eotaxin in supernatant was negatively correlated with cAMP level in ASMCs ( r = - 0. 788, P lt; 0. 01) . Conclusions There is anautocrine function in ASMCs as inflammatory cells after stimulation with sensitized serum. Eotaxin may play an important roll in the pathogenesis of asthma via a cAMP-dependent pathway.
ObjectiveTo study immunodepression effect of bone marrow-derived mesenchymal stem cell (BMSC) on acute asthmatic airway inflammation by galectin-1 (gal-1) in vivo.MethodsEighty-five female BALB/c mice were equally randomized into normal control group, asthmatic group, BMSC treatment group, gal-1 treatment group and BMSC and gal-1 inhibitor group. Ovalbumin (OVA) was used to establish acute asthmatic model. Total cell number and differential cell analysis in each group in bronchoalveolar lavage fluid (BALF) were determined. Furthermore, hematoxylin-eosin and periodic-acid Schiff staining was used to compare airway inflammation among five groups. Measurement of cytokines, including interleukin (IL) -4, IL-5 and gal-1 in BALF and OVA specific IgE (OVA-IgE) in serum were evaluated by enzyme linked immunosorbent assay. Moreover, dendritic cell (DC) in lung tissue was sorted by immunomagnetic beads and its MAPK signal pathway was analyzed by western blotting among five groups.ResultsAccumulation of inflammation cells, particularly eosinophils around airway and in BALF was evident in asthmatic mouse model, meanwhile hyperplasia of Goblet cell was also obvious in asthmatic group. BMSC engraftment or gal-1 infusion significantly reduced airway inflammation and hyperplasia of Goblet cell and the number of inflammation cells in BALF, especially eosinophils attenuated dramatically. However, there was no effect on airway inflammation and hyperplasia of Goblet Cell by simultaneous infusion BMSC engraftment and gal-1 inhibitor. Compared to normal control group, the level of IL-4, IL-5 in BALF and OVA-IgE in serum was increased remarkably in asthmatic group, but the level of gal-1 reduced obviously. Moreover, infusion of BMSC or gal-1 could mitigate the level of IL-4, IL-5 in BALF and OVA-IgE in serum and increase the level of gal-1 in asthmatic mouse. However, infusion with both BMSC and gal-1 inhibitor exerted no effect on cytokine and OVA-IgE in asthmatic mouse. DC was sorted by immunomagnetic beads and western blotting was used to detect the expression of MAPK signal pathway among five groups. The expression of ERK phosphorylation in asthmatic group was much lower than that in normal control group. On the contrary, the expression of p38 phosphorylation was much higher than that in normal control group. BMSC engraftment or gal-1 infusion significantly activated the ERK pathway and inhibited the p38 MARP pathway on asthmatic mouse DC. Nevertheless, the expression of ERK phosphorylation and p38 phosphorylation for group with BMSC and gal-1 inhibitor infusion was between the level of asthmatic group and normal control group.ConclusionsBMSC infusion alleviates airway inflammation in asthmatic mouse, especially weakens eosinophils infiltration, and the underlying mechanism might be protective effect of gal-1 secreted by BMSC which plays a role in lung tissue DC and regulates the DC expression of MAPK signal pathway.
ObjectiveTo observe repairing process of trachea epithelium cells in chlorine-induced airway epithelial injury.MethodsTwelve mice were exposed to chlorine gas and prepared the mice model of airway damage. Three mice were executed respectively on 2nd, 4th, 7th, 10th day after exposure to chlorine gas, and tracheal tissues were collected. In addition 3 normal mice served as control. Airway repair and cell proliferation were detected by EdU labeling method. The basal cell markers keratin 5 (K5), keratin 14 (K14) were adopted as the tracheal epithelial markers for locating the position of the proliferation of repairing cells. Morphological analysis was adopted to measure the proliferation rate as well as the recovery of the false stratified epithelium.ResultsIn the control group, cell proliferation rate was very low, all basal cells expressed K5, and most basal cells did not express K14. Most of epithelial cells shed from the trachea epithelium after exposure to chlorine gas. 2-4 days after chlorine exposure, K5 and K14 expression basal cells increased, K14 expression cells increased greatly. In the peak period of cell proliferation, only a small number of ciliated cells appeared in the repairing trachea area. Epithelial cells repaired fast and widely at the bottom of the trachea.ConclusionThe trachea residual basal cells play roles of progenitor cells and repair the airway epithelium after chlorine damage in mice.
Objective To investigate the effects of TiotropiumBromide on airway inflammation in a rat model of chronic obstructive pulmonary disease( COPD) . Methods Thirty Wistar rats were randomly divided into three groups. Group A received normal breeding as normal control. Group B and group C received LPS( 200 μg, intratracheally injected at the 1st and the 14th day) and tobacco exposure( from the 2nd day to the 30th day except the 14th day) to establish COPD model. And group C received a nebulized dose of Tiotropium Bromide( 0. 12 mmol / L, 10 minutes) 30 minutes before the tobacco exposure each time. Airway resistance and compliance were measured before sacrificed. Histological examination was performed with Hematoxylin-Eosin staining. The concentrations of IL-8 and LTB4 , total and differential cells counts in bronchoalveolar lavage fluid( BALF) were examined, and the concentrations of IL-8 and LTB4 in blood serum were also examined by ELISA. Results Severe lung inflammation and decreased lung function were demonstrated in the rats in the group B compared with those in the group A. The inflammatory cell counts in BALF, and the levels of IL-8 and LTB4 in BALF and serum were significantly increased in the group B compared with those in the group A. Tiotropium Bromide administration improved the parameters above. Conclusions The results suggest that Tiotropium Bromide can alleviate the lung inflammation and improve the lung function in a rat COPD model. These effects may be exerted through reducing the mediators of inflammation.
ObjectiveTo investigate the effects of prone position ventilation on oxygenation,hemodynamics and airway drainage in patients with severe aspiration pneumonia with acute respiratory distress syndrome (ARDS). Methods28 patients with severe aspiration pneumonia with ARDS admitted between January 2010 and June 2012 were recruited in the study. They were ventilated in prone position with sedation and paralysis. Mean blood pressure (MAP),heart rate (HR),central venous pressure (CVP),pulse oxygen saturation (SpO2),arterial oxygen tension (PaO2),carbon dioxide partial pressure (PaCO2),oxygenation index (PaO2/FiO2) and sputum drainage were recorded in the time points of initial supine position,prone position 1h,prone position 2h,and return to supine position 2h. ResultsCompared with the time point of initial supine position,PaO2 and PaO2/FiO2 increased significantly after 1 and 2 hours in prone position [PaO2:(85±12)mm Hg and (97±10)mm Hg vs. (65±11)mm Hg;PaO2/FiO2:(150±37)mm Hg and (158±50)mm Hg vs. (130±28)mm Hg;all P<0.05]. The effects of oxygenation improving were persistent 2h after return to supine position [PaO2:(87±11)mm Hg;PaO2/FiO2:(150±52)mm Hg,P<0.05]. There was no significant difference in MAP,HR,CVP,or SpO2 during the study. Airway sputum drainage was significantly increased 2h after in prone position compared with that in initial supine position [(15.3±2.0)mL vs. (8.1±1.1)mL,P<0.05]. Airway sputum drainage had no significant difference among 1h afer prone position,2h after return to supine position and the initial supine position [(9.1±1.0)mL and (8.3±1.2) mL vs. (8.1±1.1)mL,P>0.05]. ConclusionProne position ventilation can improve the oxygenation in patients with severe aspiration pneumonia with ARDS,and the effects of oxygenation improvement can be persistent till 2h after return to supine position. Prone position ventilation can improve sputum drainage without significant influence on hemodynamics,thus can be used as an adjuvant treatment for severe aspiration pneumonia with ARDS. The duration of prone position ventilation needs to be prolonged for patients with much airway secretion.
Respiratory oscillometry is a lung function test that measures the mechanical properties of respiratory system by the forced oscillation technique. Oscillometry can be used in those who cannot perform traditional lung function tests, including young children. It is also an important tool to assess small airways function in clinical and research fields. In 2020, the European Respiratory Society published a new technical standard for respiratory oscillometry, which offered updated technical recommendations on the hardware, software, testing protocols and quality control of oscillometry measurements. This paper interpreted the new technical standard, for providing technical suggestions regarding oscillometry measurements in clinical and research settings, and as a reference for developing technical statements and recommendations for oscillometry in China.
Objective To investigate the effects of 1, 25-( OH) 2D3 on the expression of matrix metalloprotease-9 ( MMP-9) and nuclear factor κB ( NF-κB) activity in a murine model of chronic asthma. Methods BALB/ c mice were sensitized and challenged with ovalbumin to establish chronic asthmatic model. The animals were randomly divided into a control group, an asthma group and a VD group. Lung sections from the mice were stained by HE and Masson’s trichrome, respectively. Morphometric analysis of the stained sections was performed using computerized image analysis system. Nuclear translocation of NF-κB p65 was examined using Western blot. The level of IκBαwas detected with real-time quantitative PCR ( RTPCR) and Western blot. In addition, the expression of MMP-9 in both activity and mRNA level was detected by gelatin zymograph and RT-PCR, respectively. Results Prominent airway remodeling developed in the asthma group, including the inflammatory cell infiltration, subepithelial collagen deposition and increased airway smooth muscle mass. In contrast, 1, 25-( OH) 2D3 attenuated these established structural changes of the airways. Stimulation with OVA induced a 7. 87-fold increase in the MMP-9 activity compared with that in the control group, and 1, 25-( OH) 2D3 treatment only induced a 3. 46-fold increase in the MMP-9 activity compared with that in the control group ( P lt;0. 05) . The mRNA level of MMP-9 in the VD group ( 3.16 ± 0.09) was decreased compared with the asthma group ( 5.74 ±0.13) ( P lt;0.05) , but itwas still higher than that in the control group ( 0.57 ±0.08) ( P lt;0.05) . 1, 25-( OH) 2D3 reduced the nuclear translocation of NF-κB p65 while up-regulated the IκBα level in lung tissue of chronic asthma. Conclusions 1, 25- ( OH) 2D3 can inhibit the NF-κB activity and down-regulate the expression of MMP-9 in lung tissue of chronic asthma, thus alleviating the established chronic asthma-induced airway remodeling.
Objective To invesitgate the relationship between 8-isoprostane ( 8-iso-PG) level in exhaled breath condensates ( EBCs) and severity of asthma and explore the role of 8-iso-PG in asthma evaluation and monitoring. Methods Fifty-nine patients with asthma were enrolled. In which 15 cases were acute exacerbation, 13 cases were mild intermittent, 15 cases were mild persistent, and 16 cases were moderate-to-severe persistent. Thirteen healthy volunteers were recruited as control. EBCs were collected using EcoScreen system. The 8-iso-PG levels in EBCs were measured by a specific enzyme immunoassay.The patients with mild intermittent asthma were treated with inhaled corticosteroid ( ICS) for one month and their EBCs were recollected for 8-iso-PG measurement. Results Exhaled 8-iso-PG levels were obviously increased in the patients with acute asthma compared with those chronic asthmatics [ ( 47. 2 ±6. 8) pg/mL vs ( 24. 5 ±12. 0) pg/mL, P lt; 0. 01] . In the chronic persistent asthma, the levels were significantly higher in patients with mild persistent and moderate-to-severe asthma [ ( 17. 9 ±1. 2) pg/mL and ( 39. 7 ±4. 0) pg/mL,P lt; 0. 01] . While 8-iso-PG level did not differ significantly in intermittent asthma [ ( 13. 5 ±1. 1) pg/mL]compared with the control subjects ( P gt; 0. 05 ) . After one-month ICS treatment the 8-iso-PG level in the patients with mild intermittent asthma did not change significantly although the ACT score improved. Conclusions 8-iso-PG levels in EBC are associated with the severity of asthma, implicating 8-iso-PG may be useful in monitoring airway oxidative stress in asthma. ICS treatment is incapable of decreasing the 8-iso-PG, suggesting the ICS has minor impact on oxidative stress.
Objective To explore the diagnosis and treatment of airway involvement in relapsing polychondritis. Methods The clinical data of two patients with relapsing polychondritis with airway involvement were reported and the relative literatures were reviewed. Results The two patients were both old males, with clinical manifestations of cough, dyspnea, and fever. They were misdiagnosed in a other hospital. The pulmonary function tests showed obstructive ventilatory impairemnt. On inspiratory CT, tracheal / tracheobronchial wall thickening and airway stenosis, with or without tracheal cartilage calcification were common findings. The tracheal cartilages thickeness and membranous wall were normal. On expiratory CT scans, functional abnormalities were identified such as tracheobronchomalacia. The patients were relieved by medication of corticosteroids or with immunodepressant. Conclusions The relapsing polychondritis with airway involvement is easy to be misdiagnosed. Chest CT examination is a valuable method for diagnosis of relapsing polychondritis. Corticosteroids and immunodepressant can improve the outcome.
【Abstract】 Objective To study the role of house dust mite ( HDM) induced airway epithelium TLR4 expression and T lymphocyte activation in asthmatic inflammation. Methods Thirty BALB/ c mice were randomly divided into an ovalbumin ( OVA) group, a HDMgroup, and a control group. The mice in the OVA group were sensitized with OVA and Al( OH) 3 , and repeatedly exposed to aerosolized OVA. The mice in the HDMgroup and the control group were sensitized and challenged with HDMand saline, respectively.Histopathology changes of pulmonary tissue and airway were observed under light microscope. Levels of IL-4, IL-5, IL-13, IL-17, and IFN-γin BALF were measured by ELISA. Total and differential cell counts in bronchoalveolar lavage fluid ( BALF) were also measured. The mRNA and protein expressions of TLR4 weredetected by quantitative real-time PCR and Western blot, respectively. Th1, Th2, and cells in the peripheral blood were detected by flow cytometry. Results Light microscope revealed eosinophil specific inflammatory cells infiltration around the peribronchovascular region,mucus gland hyperplasia, and airway mucous plug inthe OVA group. The HDM group showed more severe alveolar and intersititial congestion and neutrophils infiltration. The control group showed intact alveolus with few mucous plug and inflammatory cells.Compared with the OVA group, significant increases in the number of total cells and neutrophils, as well as significantly higher expression of IL-4, IL-5, IL-13, and IL-17 were detected in the HDMgroup ( P lt;0. 05) ,while IFN-γexpression had no significant change ( P gt;0. 05) . The expression of TLR4 mRNA and protein significantly increased in the HDMgroup( P lt; 0. 05) , and did not change significantly in the other two groups ( P gt;0. 05) . The percentages of Th2 and Th17 cells in peripheral blood in the HDMgroup were significantly higher than the OVA group ( P lt;0. 05) . Conclusion HDM may induce inflammatory cells infiltration andactivation of Th2 and Th17 lymphocyte cells via up-regulation of TLR4 expression in airway epithelium,which might play an important role in asthmatic inflammation.