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.
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 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.
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 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.
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 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 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．
【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 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.