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 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 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.
【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 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.
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 explore the role of nuclear factor kappa B(NF-KB)in the pathogenesis of chronic obstructive pulmonary disease(COPD)and the therapeutic efects of glucocorticoid．Methods Twenty-four Wistar rats were randomly divided into three groups，ie．normal control，COPD model and prednisone preventive treatment group．Rat COPD model Was established by exposing the rats to cigarette smoke daily．Prednisone Was given through stomachal injection on altemate days．After COPD model Was set up，bronchoalveolar lavage(BAL)Was performed．Total cell counts and neutrophil counts in BALF were examined．Pathological changes of lung tissue Was observe0 by hematoxylin-eosin staining．The morphological indices of pulmonary emphysema(MLI，MAN and PAA)Was measured by a computerizedimage analyzer and compared in three groups．NF-KB expression in lung tissues were detected by immunohistochemistry assay．Rults Emphysema Was confirmed by three morphological indices in COPD model group compared to those of normal control group[MLI：(97.97±11.10)×10-6m vs (47．23±2．80)×10-6 m，MAN：(95.98±l4.89)×106 ／m vs (164.21±9．30)×106 ／m ，PAA：(64 ±5.7)％vs (44±2.7)％，Plt;0.01]．Total cell counts and neutrophil counts in BALF of COPD model group were significantly higher than those of control group[（5.76±0.29）×108/L vs （1.64±0.12）×108/L，（1.26±0.25）×108/L vs （0.099±0.065）×108/L，Plt;0.01]．After the preventive treatment with prednisone，MLI，MAN and PAA were significantly changed[（57.66±4.62）×10－6mvs （97.97±11.10）×10－6m，（111.40±16.92）×106个/m2 vs （95.98±14.89）×106个/m2，Plt;0.01；（58±6.1）% vs （64±5.7）%，Plt;0.05]，which indicated that airway inflammation and emphysematous injury in preventive treatm ent group were milder than those of COPD mode1．Total ceil counts and neutrophil countsin BALF were found in preventive treatment group as compared to those of COPD model[[（3.18±0.29）×108/L vs （5.76±0.29）×108/L，（0.57±0.12）×108/L vs （1.26±0.25）×108/L，Plt;0.01].The percentage of positive cells of NF-KB nuclear staining in bronchiolar epithelial ceils was significantly increased in the COPD group than that in the control group[（29.02±1.25）% vs （12.17±1.13）%，Plt;0.01]，but was significantly decreased in the preventive treatment group[(19.23±1.18)％vs (29.02±1.25)％，Plt;0．01]．Conclusions NF-KB may be responsible for the persistence and amplification of inflammation in COPD through neutrophil recruitment and activation．Prednisone may suppress airwayinflammation in COPD by inhibiting NF-KB．
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.
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.
0bjective To study the effect of bacterial infection on acute exacerbation of chronic obstructive pulmonary disease(AECOPD)，and to compare the airway inflammation caused by different isolated bacteria．Methods A total of 159 sputum samples were collected from AECOPD patients diagnosed according to GOLD 2004 standard，in which conventional culture and identification of bacteria was conducted．The patients with purulent sputa were divided into different groups according to bacteria separated．Levels of IL-6．IL-8 and TNF-α in sputum supernatant were assayed and compared in different bacteria groups．the purulent sputum without isolated bacteria group(NG)and normal control group(NC)． Results One hundred and twenty-nine strains of bacteria were isolated in 159 qualified sputa，including 26 strains of Klebsiella pneumoniae(KB)，21 strains of Hemophilus influenza(Hi)，17 strains of Pseudomonas aeruginosa(PA)，37 strains of Haemophilus parainflb~enzae(HP)(mixed infection not included)and 28 strains of other bacteria．Among of all samples，20 were double infection of Haernophilus parainfluenzae with another bacterium．Ninety-seven purulent sputa were collected．According to bacteria isolated，these sputa were divided into five groups，named HP(24 samples)，Hi(20 samples)，PA(16 samples)，KB(19 samples)and NG(18 samples)．Contrast to NC，concentration of IL-8 and TNF-α rose in sputa from which PA，Hi，KB were isolated(Plt;0．05)．The level increased much more in sputa from which PA and Hi were isolated compared with KB(Plt;0．05)．Concentration of IL-6 rose in sputa of each group collected from AECOPD patients contrast to sputa collected from NC(Plt;0．05)，without significant differences among all other group except for NC．Conclusions Bacterial infection plays an important role in AECOPD characterized with high level of inflammatory factors especially when PA，Hi，KB were infected bacteria．This study provides evidence for antibacterial therapy in AECOPD patients．