Objective To investigate the expression of stromal cell derived factor-1 ( SDF-1) and the effects of budesonide suspension for inhalation ( Pulmicort Respules) in mice with asthma. Methods Thirty Kunming female mice were randomly divided into three groups, ie. a control group, an asthma group, and a pulmicort treatment group. The asthma group and the pulmicort treatment group were sensitized with ovalbumin ( OVA) by a combination of intraperitoneal injection and repeated OVA intranasal challenges to establish mouse asthma model. The pulmicort treatment group received 100μL pulmicort by intranasal administration before OVA challenge. The immunohistochemistry was used to estimate the expression of SDF-1 in lung tissues. HE staining and Wright-Giemsa staining method were used to assess inflammatory infiltration in the airway and bronchoalveolar lavage fluid ( BALF) respectively. Results The expression of SDF-1 in the asthma group increased significantly compared with the control group ( 0.48 ±0.03 vs. 0.21 ± 0.02, Plt;0.05) , and significantly decreased after the intervention with pulmicort ( 0.29 ±0.01 vs. 0.48 ± 0.03, Plt; 0.05 ) . Compared with control group, the infiltration of inflammatory cells in airway was significantly enhanced in the asthma group, and attenuated in the pulmicort treatment group. The total number of inflammatory cells and eosinophil, lymphocyte, neutrophil counts in BALF increased significantly in the asthma group compared with the control group, and decreased significantly after pulmicort intervention. Conclusion SDF-1 may play an important role in the recruitment of inflammatory cells in asthmatic airway and pulmicort may relieve airway inflammation by decreasing the expression of SDF-1.
ObjectiveTo monitor the airway inflammatory factors in exhaled breath condensate(EBC) of severe stable COPD patients during salmeterol/fluticasone (50/500μg, bid) treatment, and explore their clinical significance. MethodsTwenty-four sever stable COPD patients and 18 healthy controls were included in the study. EBC was collected from COPD patients before treatment (day 0) and 14 days, 28 days, 90 days after treatment. Meanwhile lung function test and SGRQ score were measured.Concentrations of IL-6 and IL-10 were measured by liquid chip and 8-isoprostane by enzyme-linked immunosorbent assay. ResultsLevels of 8-isoprostane, IL-6 and IL-10 in EBC were significantly higher in the sever stable COPD patients before treatment compared with the healthy controls. 8-isoprostane was decreased significantly at day 14 compared with day 0[(11.59±4.12) pg/mL vs. (14.17±4.66) pg/mL, P < 0.05], and kept in low level till day 90 (P > 0.05). IL-6 was significantly decreased at day 28 compared with day 0[(1.46±0.19) pg/mL vs. (1.59±0.19) pg/mL, P < 0.05], but did not change significantly till day 90. IL-10 was in low level but showed increase at day 90 compared with day 28[(1.72±0.19) pg/mL vs. (1.62±0.12) pg/mL, P < 0.05]. FEV1 and FEV1/FVC were improved and SGRQ score was decreased after 90 days treatment (P < 0.05). FEV1 was not correlated with 8-isoprostane, IL-6 or IL-10 level. ConclusionsDynamic observation of EBC 8-isoprostane level in severe COPD patients can help in evaluating drug efficacy. IL-10 may play a role in airway anti-inflammation.
【Abstract】 Objective To investigate the effect of allogeneic bone marrow-derived mesenchymal stem cells ( BMSCs) transplantation on the airway inflammation and airway remodeling in chronic asthmatic mice. Methods Forty female BALB/c mice were equally randomized into four groups, ie. a normal control group, a BMSCs control group, an asthma model group, and a BMSCs transplantation group. BMSCs were generated from male donor mice, then the mice in the asthma model group and the BMSCs transplantation group were sensitized and challenged with OVA to establish chronic asthmatic mice model. Hematoxylin and eosin staining and Alcian blue-periodic acid-Schiff staining were used to analyze the effects on airway inflammation and airway remodeling after BMSC engraftment. The number of CD4 + CD25 + regulatory T cells in spleen was detected by flow cytometry. Results In lungs of the asthmamodel group, there were intensive inflammatory cells infiltration around airway and blood vessels, goblet cell proliferation, epithelial desquamation, patchy airway occlusion by hyperviscous mucus, and hypertrophy of airway smooth muscle.Airway inflammation and airway remodeling were significantly relieved in the BMSCs transplantation group.There was no obvious inflammatory cells infiltration in the airway and airway remodeling both in the normal control group and the BMSCs control group. The number of CD4 + CD25 + regulatory T cells in spleensignificantly decreased in the asthma model group compared with the two control groups ( P lt; 0. 05) , and significantly increased in the BMSCs transplantation group compared with the asthma model group ( P lt;0. 05) . There was no significant difference in the number of CD4 + CD25 + regulatory T cells in spleen betweenthe control groups and the BMSCs transplantation group. Conclusion BMSCs engraftment can up-regulate CD4 + CD25 + regulatory T cells and relieve airway inflammation and airway remodeling in asthmatic mice.
Objective To investigate the effect of myeloid derived suppressor cells ( MDSCs) on airway inflammation of asthmatic mice. Methods Five male BALB/ c mice aged 6 weeks were used for preparing 4T1 tumor bearing mice. Thirty female BALB/ c mice aged six weeks were randomly divided into a normal control group, an athmatic model group, and a cell transplantation group. The MDSCs were separated frommyeloid tissue of tumor-bearing mice using amagnetic cell sorting systemand cultured in RPMI medium 1640 containing GM-CSF. The morphologic characteristics of these cells were observed under lightmicroscope and the phenotypic figures were analyzed with flow cytometry. The mice in the model group and the cell transplantation group were sensitized by ovalbumin and then stimulated with nebulized ovalbumin. The mice in the cell transplantation group were intravenously administered MDSCs which purified by magnetic cell sorting system at 10 days after sensitization. The airway inflammation was evaluated by HE staining. The total and differential cell counts in bronchoalveolar lavage fluid ( BALF) were measured.Results The neutrophil and eosinophil infiltration in pulmonary tissue was dramatically increased in the model group, but not observed in the normal control group and was much milder in the cell transplantation group. The total cell count, the eosinophil and lymphocyte counts in BALF of the model group and the cell transplantation group were significantly higher than those of the normal control group( P lt; 0. 05) , and the number of eosinophils in BALF of the cell transplantation group was decreased when compared with that of the model group( P lt;0. 05) . Conclusion MDSCs via intravenous infusion can effectively suppress airway inflammation in a mouse asthma model.
Objective To observe the effects of astaxanthin (AST) on the airway inflammation and remodeling in the asthmatic rats. Methods Fifty male Wistar rats were randomly divided into five groups (n=10 for each group): saline-sensitized and-saline-challenged group (the control group), bronchial asthma group (the asthma group), bronchial asthma+astaxanthin 5 mg/kg gavage treatment group (the AST 5 mg/kg group), bronchial asthma+10 mg/kg gavage treatment group (the AST 10 mg/kg group), and bronchial asthma+50 mg/kg gavage treatment group (the AST 50 mg/kg group). The level of interleukin-5(IL-5), interleukin-13(IL-13), interferon-γ(IFN-γ), tansforming growth factor-β (TGF-β), malondialdehyde (MDA) and superoxide dismutase (SOD) in the bronchoalveolar lavage fluid (BALF) and the total IgE level in the serum were measured using enzyme linked immunosorbent assay (ELISA).The infiltration of airway inflammatory cells and the degree of airway epithelial cells detachment, the extent of goblet cell hyperplasia and the severity of subepithelial collagen deposition were evaluated on the hematoxylin eosin (HE), periodic acid Schiff (PAS) and Masson trichrome stained lung sections. reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of mucin 5A and C (MUC5AC) messenger ribonucleic acid(mRNA) in lung tissue; Immunohistochemical staining was used to determine the expression of MUC5AC protein in the rat airway epithelium. Results The level of IL-5, IL-13, TGF-β, MDA and the total IgE in the serum respectively [(36.73±2.29), (53.99±2.70), (60.89±2.54)ng/mL,(18.65±0.76)umol/L, (54.50±2.91)ng/mL], the extent of inflammatory cells infiltration (46.24 ± 4.26), the extent of eosinophils infiltration (2.09± 0.13), the extent of epithelial cells detachment [(6.09±0.45)%], the extent of goblet cell hyperplasia [(13.65±1.90)%], the extent of subepithelial collagen deposition [(17.58±2.14)%], the MUC5AC mRNA expression level, and the lung tissue MUC5AC protein expression IOD value (187±12) in the asthma group were all higher than those in the control group (P<0.01 or P<0.001), the level of IFN-γ and SOD in the BALF[(26.38±1.70) ng/mL], [(16.37±1.22) U/L], was lower than that in the control group (P<0.001); The level of IL-5, IL-13, total IgE, TGF-β, MDA, the inflammatory cells infiltration in the airway epithelial, the degree of epithelial cell damage and detachment, the degree of goblet cell hyperplasia, the degree of subepithelial collagen deposition, the MUC5AC mRNA expression in lung tissue,and the MUC5AC protein expression in airway epithelial cells in the AST treated groups were all lower than those in the asthma group (P<0.05 or P<0.01 or P<0.001),the level of IFN-γ, SOD in the BALF was higher than that in the asthma group (P<0.05 or P<0.01). Conclusion Astaxanthin can inhibit airway inflammation, downregulate airway MUC5AC expression, inhibit goblet cell proliferation, and alleviate airway remodeling in rats with bronchial asthma.
ObjectiveThrough measuring fractional exhaled nitric oxide (FeNO) and eosinophil levels of peripheral blood in chronic obstructive pulmonary disease (COPD) patients with different phenotype of acute exacerbation frequency, to predict the therapeutic effect of glucocorticoid therapy and guide the clinical treatment of different subtypes patients with acute exacerbations of COPD.MethodsA total of 127 patients with acute exacerbation of COPD in Suining Central Hospital from February 2017 to October 2019 were recruited. They were divided four groups according to the number of acute exacerbations in the past one year and the treatment scheme, ie. a frequent acute exacerbation with glucocorticoid treatment group (34 cases), a frequent acute exacerbation with non-glucocorticoid treatment group (31 cases), a non-frequent acute exacerbation with glucocorticoid treatment group (30 cases), and a non-frequent acute exacerbation with non-glucocorticoid treatment group (32 cases). FeNO value, eosinophil ratio in peripheral blood, COPD assessment test (CAT) score, and interleukin-8 (IL-8) concentration were measured before and on the 10th day of treatment, and the differences within group and between groups before and after treatment were compared.ResultsCAT score, FeNO, eosinophil ratio and IL-8 level in the four groups were significantly improved on the 10th day after treatment (all P<0.05). The declines of FeNO value, eosinophil ratio, and IL-8 level on the 10th day of treatment compared with those before treatment in the frequent acute exacerbation with glucocorticoid treatment group and the frequent acute exacerbations with non-glucocorticoid treatment group were larger than those in the non-frequent acute exacerbation with glucocorticoid treatment group and the non-frequent acute exacerbation with non-glucocorticoid treatment group (all P<0.05). The declines of FeNO value, blood eosinophil ratio and IL-8 level in the frequent acute exacerbation with glucocorticoid treatment group were also statistically significantly larger than those in the frequent acute exacerbations with non-glucocorticoid treatment group (all P<0.05). The improvement of CAT score in the frequent acute exacerbation with glucocorticoid treatment group was greater than that in other three groups (all P<0.05). There was no significant difference in CAT score between the non-frequent acute exacerbation with glucocorticoid treatment group and the non-frequent acute exacerbation with non-glucocorticoid treatment group (P>0.05).ConclusionsThe degree of airway inflammation is more obvious in patients with frequent acute exacerbation phenotype of COPD. FeNO value can reflect the level of airway inflammation in patients with frequent acute exacerbation of COPD and evaluate the response to glucocorticoid therapy.
ObjectiveTo investigate the synergistic effect of cold stress plus particulate matter 2.5 (PM2.5) co-exposure on the occurrence of respiratory inflammation and the possible post-transcriptional regulation mechanism of cold inducible RNA-binding protein (CIRP).MethodsIn vivo and in vitro experiments were carried out, and the lung tissue specimens from human surgical resection were observed. The rat model and cultured airway epithelial cells 16HBE were respectively divided into four groups (n=8), namely blank control group, 5 °C/18 °C group, PM2.5 group and 5 °C/18 °C+PM2.5 group. The expression of mRNA and protein of representative inflammatory cytokines and CIRP of cultured airway epithelial cells and rat bronchial/pulmonary tissues were respectively detected by ELISA, qPCR, and Western blot. Furthermore, the temporal dynamics of CIRP distribution were observed by cellular immunofluorescence. Finally, immunohistochemical method was used to observe the localization and expression of CIRP in rat and human bronchial/pulmonary tissues at the same time.ResultsIn vivo experiments, the mRNA and protein expression levels of CIRP, interleukin-6, and tumor necrosis factor-α in 5 °C group and PM2.5 group were significantly higher than those in the control group (all P<0.05), while the expression level of mRNA and protein in 5 °C+PM2.5 group were increased most obviously (all P<0.01). The same rule also appeared in the experimental results of each group in the vitro experiment. In addition, CIRP was mainly located in the cell nucleus; compared with the control group, the intracellular shift of CIRP appeared in 18 °C group and PM2.5 group, while the migration phenomenon was most obvious in the 18 °C+PM2.5 group. In the immunohistochemistry of rat bronchus/pulmonary tissue, the expressions of CIRP in the 5 °C group and in the PM2.5 group were significantly higher than those in the control group, and the CIRP expression in 5 °C+PM2.5 group was increased most evidently. Moreover, CIRP was expressed in the bronchial epithelial mucosa of normal people and patients with chronic obstructive respiratory disease (COPD), and it is mainly located in the nucleus of airway mucosal epithelial cells. The CIRP expression of COPD patients was significantly higher than that in the normal population.ConclusionCold stress has a sensitizing effect on airway epithelial inflammatory response induced by PM2.5, and post-transcriptional regulation of CIRP translocation from nucleus to cytoplasm may be an important mechanism.
Objective To explore the effect of lower airway inflammation on the pathogenesis of upper airway cough syndrome( UACS) . Methods Ten cases of UACS and 10 cases of chronic rhinitis or sinusitis without cough were enrolled as group A and group B, respectively. And 10 healthy volunteers were included as controls( group C) . The cough threshold C2 and C5 to inhaled capsaicin, defined as the lowest concentration of capsaicin required to induce ≥2 and ≥5 coughs, was measured. The total and differential cell counts was determined in induced sputum, and the levels of histamine and prostaglandin E2 were analyzed in supernatant of sputum. Results Cough threshold was significantly lower in group A than group B [ C2: ( 0.65 ±0. 08) μmol / L vs ( 3.90 ±1. 37) μmol / L; C5: ( 1.59 ±0. 28) μmol / L vs ( 33.46 ±23. 71) μmol / L, P lt;0. 05] and comparable between group B and group C( P gt; 0. 05) . Group A, similar to group B( P gt; 0. 05 ) , contained more inflammatory cells, with decreased percentage of macrophages and increased percentage of neutrophils in induced sputum than group C( P lt; 0. 05) . Furthermore, the levels of histamine[ ( 9. 55 ±1. 89) ng/mL vs ( 2. 37 ±0. 25) ng/mL, P lt; 0. 05] and prostaglandin E2 [ ( 361. 71 ±39. 38) pg/mL vs ( 144. 34 ±15. 69) pg/mL, P lt; 0. 05] were higher in supernatant of induced sputum from group A than group B, while the latter was not different from group C( P gt; 0. 05) . Conclusion Increased cough sensitivity caused by airway inflammation may be important for the pathogenesis of UACS, and the activation of mast cells in mucosa of lower airway might be an important factor.
Objective To investigate the modulating roles of Clara cell secretory 16 kD protein ( CC-16) , transcription factor T-bet, and GATA-3 in airway inflammation of patients with asthma. Methods 25 patients with acute exacerbation of asthma were enrolled as an asthma group and 33 healthy volunteers were enrolled as control. The plasma levels of CC16, IFN-γ, and IL-4 were measured by enzyme-linked immunosorbent assay ( ELISA) . The mRNA expressions of T-bet and GATA-3 in the peripheral bloodmononuclear cells ( PBMCs) were measured by reverse transcription-polymerase chain reaction ( RT-PCR) .Results The levels of CC16 and IFN-γin the asthma group were lower than those in the control group [ ( 21. 96 ±7. 31 ) ng/mL vs. ( 64. 88 ±25. 27) ng/mL, ( 118. 73 ±22. 59) pg/mL vs. ( 145. 53 ±29. 50) pg/mL, both P lt;0. 01] . The IL-4 level in the asthma group was significantly higher than that in the control group [ ( 425. 22 ±4. 37) pg/mL vs. ( 69. 72 ±10. 15 ) pg/mL, P lt; 0. 01] . The T-bet mRNA expression and T-bet /GATA-3 ratio of PBMCs in the asthma group were significantly lower than those in the control group( both P lt; 0. 01) . The expression GATA-3 mRNA was significantly higher than that in the control group( P lt;0. 01) . The level of CC16 was positively correlated with T-bet mRNA expression and the ratio of T-bet /GATA-3 ( r =0. 792, 0. 761, respectively, P lt; 0. 01) . There was no correlation between CC16 and the GATA-3 mRNA expression ( P gt;0. 05) . Conclusions These results suggest that CC16 and T-bet play important protection roles in the pathogenesis of asthma. GATA-3, IFN-γ, and IL-4 also participate in the airway inflammation of asthma.
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.