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.
ObjectiveExploring the potential causal effects and directions of insulin resistance (IR) and chronic airway inflammatory diseases, including asthma and chronic obstructive pulmonary disease (COPD), through two sample Mendelian randomization (MR). MethodsA total of 53 validated single nucleotide polymorphisms (SNPs) associated with IR were selected as instrumental variables. The inverse variance-weighted (IVW) method was used to model the causal association, and sensitivity analyses through leave-one-out analysis and pleiotropy testing were conducted to assess the relationship between IR and asthma and COPD. ResultsMR analysis revealed no significant causal effect of IR on asthma (IVW: OR=1.067, 95%CI 0.871 to 1.306, P=0.531) or COPD (IVW: OR=0.906, 95%CI 0.686 to 1.196, P=0.557). The results were consistent across sensitivity analyses and multiple pleiotropy tests, with no evidence of horizontal pleiotropy detected. ConclusionNo causal association was found between IR and the development of asthma or COPD. The relationship between these conditions may be influenced indirectly through complex interactions between metabolic and inflammatory pathways affecting disease progression.
Objective To investigate the effects of FasL gene-modified dendritic cell (DC) on the airway inflammation in mice sensitized/challenged by house dust mite (HDM) allergen.Methods FasL gene-modified DC (FasL-DC) and control DC (nontransfection DC) were administrated into HDM sensitized and challenged mice by intratracheal injection respectively，then HDM sensitized and challenged mice were sacreificed two days later.Total and differentiation cell counts and levels of interleukin-4（IL-4），IL-5 and interferon-γ(IFN-) in bronchoalveolar lavage fluid (BALF) were detected and lung histological features were observed.Results After administration of FasL-DC，lung allergic inflammation was ameliorated while total cell counts，the percentage of eosinophil ,the levels of IL-4 and IL-5 in BALF decreased and the level of IFN- in BALF increased.Conclusion Administrating FasL-DC into HDM sensitized/challenged mice can inhibit Th2 cells activation and ameliorate airway allergic inflammation.
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.
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.
ObjectiveTo investigate the expressions of IL-10,tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in serum and lung tissue of COPD rats in order to elucidate the potential mechanism of airway inflammation. MethodsForty-five healthy adult male SD rats were randomly divided into a COPD model group (n=30) and a normal control group (n=15). The COPD rat model was established by intratracheal instillation of lipopolysaccharide (LPS) and exposure to cigarette smoke for 28 days. The concentrations of IL-10,TNF-α and IFN-γ in serum and lung tissue were measured by ELISA. ResultsTNF-α level of serum and lung tissue in the COPD model group increased significantly compared with the control group(P<0.05),while the levels of IFN-γ and IL-10 decreased significantly[serum:(44.68±8.67) ng/L vs. (75.96±10.59) ng/L;lung tissue:(64.55±9.03) ng/L vs. (94.06±8.71) ng/L,P<0.01]. The level of IL-10 in serum and lung tissue was negatively correlated with TNF-α (serum:r=-0.67,lung tissue:r=-0.80,P<0.01). The level of IL-10 in serum and lung tissue was positively correlated with IFN-γ (serum:r=0.64,lung tissue:r=0.72,P<0.01). The level of IL-10 in serum and lung tissue was negatively correlated with the percentage of neutrophils(serum:r=-0.70,lung tissue:r=-0.67,P<0.01). ConclusionIn COPD rats,down regulation of IL-10 plays an important role in regulation of airway inflammation.
Objective To observe the effect of NLRP3 inflammasome inhibitor MCC950 intervention on airway Muc5ac level in asthmatic mice, and to explore the role and mechanism of NLRP3 inflammasome in asthmatic airway mucus hypersecretion. Methods A total of 50 SPF grade BALB/c female mice aged 6 - 8 weeks were randomly divided into normal control group (NS group), asthma model group (AS group), dexamethasone group (Dex group), MCC950 high-dose intervention group (MH group) and MCC950 low-dose intervention group (ML group), with 10 mice in each group. Furthermore, the bronchoalveolar lavage fluid (BALF) of mice in each group was counted by total cell count, associated with white blood cell different count. In addition, the concentrations of interleukin (IL)-18 and IL-1β in BALF were tested by enzyme-linked immunosorbent assay; The lung tissues were prepared into paraffin-embedded sections, which were then subject to hematoxylin-eosin (HE) staining, Alcian blue-periodic acid Schiff base staining and Masson staining to observe the pathological changes of lung tissues. Immunohistochemistry was used to detect the protein expression levels of Muc5ac, NLRP3 and caspase-1 in lung tissues. Real-time quantitative polymerase chain reaction was performed to detect the relative mRNA expressions of Muc5ac, NLRP3 and Caspase-1 in lung tissues. Results Compared with NS group, AS group showed significant increase in total cell count of BALF, the percentage of eosinophils, the infiltration score of inflammatory cells around the airway, the positive relative staining area of airway mucus and the deposition area of airway collagen fibers in mice (P<0.05), upregulated protein expression levels of Muc5ac, NLRP3 and Caspase-1 in lung tissues (P<0.05), elevated relative mRNA expressions of Muc5ac, NLRP3 and Caspase-1 in lung tissues (P<0.05), and raised concentrations of IL-18 and IL-1β in BALF (P<0.05). While compared with AS group, the above indicators were reduced in MH group and ML group (P<0.05). Moreover, in relative to Dex group, these indicators were increased in MH group ML group (P<0.05). In addition, no statistically significant difference was observed in the aforementioned indications between MH group and ML group.Conclusions MCC950 intervention can inhibit airway inflammation and airway mucus secretion in asthmatic mice. Its mechanism is speculated to be related to the suppression of NLRP3, Caspase-1, IL-18 and IL-1β expressions, downregulation of Muc5ac expression, and inhibition in airway mucus hypersecretion.
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.
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.