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 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．
Objective To explore the effects of prolonged Aspergillus fumigatus spores inhalation on airway inflammation and remodeling in rats with chronic obstructive pulmonary disease（COPD）.Methods Fifty Wistar rats were randomly divided into group A，B，C，D and E，(n=10 in each group) and group E was served as normal control.In group A，B，C and D，COPD models were established by intratracheal administration of lipopolysaccharide (LPS) combined with cigarette smoke exposure.The rats in group A，B and C were given intranasal inhalation of 1×106cfu spores，1×103cfu spores and 100 mL saline twice a week for consecutive 5 weeks，respectively，while the rats in group D were given no treatment.Bronchoalveolar lavage fluid(BALF) were collected for total and differential cell count，and interleukin-8（IL-8） and transforming growth factor-b(TGF-b) concentration measurement.The pathologic changes of lung tissue were observed by HE，PAS and Masson stainings.Results Pathological changes characteristic of COPD were found in group D.The total cell count，the percentage of neutrophile and lymphocyte in BALF in group A and B were higher than those in group C and D(all Plt;0.01).IL-8 and TGF-b in BALF in group A and B were higher than those in group C and D(all Plt;0.01).The pathologic score of airway inflammation in group A was higher than those in group B，C and D(all Plt;0.01)：The thickness of airway wall(WAt/Pbm) and airway smooth muscles(WAm/Pbm)，the collagen deposition in the total airway wall(WCt/Pbm) and in the outer airway wall(WCo/Pbm) and the percentage of goblet cells to epithelial cells in group A and B were higher than those in group C and D(all Plt;0.01).In group A and B，IL-8 was positively correlated with the percentage of neutrophile(r=0.856，Plt;0.01)，the pathologic score of airway inflammation(r＝0.884，Plt;0.01)，and the percentage of goblet cells to epithelial cells (r=0.702，Plt;0.05)，respectively.TGF-b was positively correlated with WAt/Pbm，WCt/Pbm，WCo/Pbm and the ratio of goblet cells to epithelial cells (r=0.706，Plt;0.05：r=0.802，Plt;0.01：r=0.876，Plt;0.01：r=0.713，Plt;0.05).Conclusion Prolonged inhalation of Aspergillus fumigatus spores can aggravate the airway inflammation and remodeling in rats with COPD.
Objective To investigate the role of endogenous Hydrogen Sulfide ( H2S) in airway inflammation and responsiveness in a rat model of chronic passive-smoking. Methods Male SD rats were randomly divided into a control group ( breathing fresh air) and a passive smoking group [ cigarette smoking( CS) passively] , with 18 rats in each group. Six rats in each group were randomly intraperitoneally injected with normal saline, sodium hydrosulfide ( NaHS) or propargylglycine ( PPG, an irreversible inhibitor of cystathionine- γ-lyase) . The animals were divided into six subgroups, ie. Con group, NaHS group, and PPG group, CS group, CS+ NaHS group, and CS + PPG group. After 4 months, lung histological change and airway tension were measured. The H2S levels of plasma and lung tissue were analyzed by the sensitive sulphur electrode assay. The expression of cystathionine-γ-lyase ( CSE) was measured by western blot. Results Compared with the Con group, CSE protein expression in lung tissues was increased in CS group( P lt;0. 05) ; the H2 S levels of plasma were significantly higher in CS group, NaHS group and CS + NaHS group, and much lower in PPG group ( P lt; 0. 05, respectively) . Compared with CS group, the H2S levels of plasma were significantly higher in CS + NaHS group, and much lower in CS + PPG group( P lt; 0. 05, respectively) . The H2S level of lung tissue in each group had no significant difference ( P gt; 0. 05) . Compared with Con group,score of lung pathology was significant elevated, and the responsiveness of airway smooth muscles to ACh and KCl was significant augmented in CS group. Compared with CS group, the score of lung pathology was decreased, and the responsiveness of airway smooth muscles was decreased in CS +NaHS group( P lt;0. 05) , and vise versa in CS + PPG group( P lt; 0. 01) . Conclusion H2S can alleviate airway inflammation and hyperresponsiveness induced by CS, and administration of H2S might be of clinical benefit in airwayinflammation and airway responsiveness.
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 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.
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.
【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 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.