Objective To observe the eotaxin expression of rat airway smooth muscle cells ( ASMCs) induced by serum from asthmatic rats, and explore the possible mechanism. Methods ASMCs isolated fromrat tracheas were cultured in vivo. Then they were treated with serum from asthmatic rats, or treated with serum and dexamethasone simultaneously. The level of eotaxin protein in supernatant and eotaxin mRNA in ASMCs were measured by ELISA and reverse transcription-polymerase chain reaction. The expression of cAMP in ASMCs was examined by radioimmunoassay. Results After the treatment with sensitized serum, the eotaxin level in supernatant and mRNA expression in ASMCs were significantly higher [ ( 107. 09 ±7. 12) ng/L vs. ( 0. 63 ±0. 56) ng/L, P lt; 0. 05; 1. 39 ±0. 04 vs. 0. 05 ±0. 01, P lt;0. 05] , and the level of cAMP in ASMCs was significantly lower compared with the control group [ ( 17. 58 ±3. 62) ng/L vs. ( 32. 39 ±3. 36) ng/L, P lt; 0. 05] . After intervened by the sensitized serum and dexamethasone simultaneously, the protein and mRNA expressions of eotaxin were lower compared with those intervened by sensitized serumalone [ ( 64. 18 ±4. 04) ng/L and 0. 77 ±0. 19] . The level of eotaxin in supernatant was negatively correlated with cAMP level in ASMCs ( r = - 0. 788, P lt; 0. 01) . Conclusions There is anautocrine function in ASMCs as inflammatory cells after stimulation with sensitized serum. Eotaxin may play an important roll in the pathogenesis of asthma via a cAMP-dependent pathway.
Objective To explore the effects of prolonged inhalation of Aspergillus fumigatus ( Af)spores on epithelial cell injury and expression of epidermal growth factor receptor( EGFR) in airways of asthmatic rats. Methods 64 male Wistar rats were randomly divided into 8 groups, ie. chronic asthma group ( group A) , chronic asthma plus Af spores inhalation for 1 week ( group B) , 3 weeks ( group C) and 5 weeks ( group D) , chronic asthma plus saline inhalation for 5 weeks ( group E) , OVA-sensitized and salinechallenged group ( group F) , and OVA-sensitized and saline-challenged plus Af spores inhalation for 5 weeks ( group G) ( each n =8) . The airway resistance ( Raw) and change of Raw after acetylcholine provocation were detected using a computerized system. The concentrations of epidermal growth factor ( EGF) andtransforming growth factor alpha( TGF-α) in BALF were measured by ELISA. The extents of epithelial cell injury and goblet cell hyperplasia were evaluated on hematoxylin and eosin-stained( HE) and periodic acidschiff ( PAS) stained lung sections. The expression of EGFR in airway epithelia was demonstrated byimmunohistochemistry, and the level of EGFR protein in the rat lung tissues was measured by western blot.Results The concentration of EGF( pg/mL) ( 51. 72 ±8. 54, 68. 12 ±7. 85, 86. 24 ±9. 12, respectively)and TGF-α( pg/mL) ( 55. 26 ±9. 30, 75. 58 ±11. 56, 96. 75 ±14. 66, respectively) , detached/ inner perimeter of epithelium( % ) ( 11. 25 ±3. 12, 26. 45 ±5. 56, 28. 50 ±7. 50, respectively) , the ratio of goblet cell area to epithelial cell area ( % ) ( 16. 42 ±5. 24, 22. 64 ±6. 82, 36. 38 ±9. 21, respectively) , the integrated optical density ( IOD) of EGFR positive stain in airway epithelial cells ( 82 ±15,120 ±19, 165 ±21, respectively) , and the EGFR protein levels in lung tissues ( 0. 91 ±0. 26, 1. 61 ±0. 52, 2. 52 ±0. 78,respectively) in group B, C, and D were higher than those in group A, E, F and G( P lt; 0. 05 or P lt;0. 01) .The change rates of Raw( % ) ( 61. 91 ±5. 26, 84. 69 ±6. 38) in group C and D were higher than those in group A, E, F and G ( P lt; 0. 05 or P lt;0. 01) . The IOD of EGFR was positively correlated with detached/inner perimeter of epithelium( % ) and the ratio of goblet cell area to epithelial cell area( % ) ( r = 0. 692,P lt;0. 01; r = 0. 657, P lt; 0. 01, respectively) . Conclusion Prolonged inhalation of Aspergillus fumigatus spores can aggravate airway epithelial cell injury, up-regulate the expression of EGFR in airway epithelial cell and induce goblet cell hyperplasia, thus increase the airway responsiveness in rats with chronic asthma.
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個/m2 vs （95.98±14.89）×106個/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 identify the short ( lt;30 days) and intermediate ( 30 days to 6 months) benefits and risks of tracheobronchial stents in patients with malignant airway stenosis. Methods 55 cases with malignant airway disease who underwent tracheobronchial stents placement from January 2006 to May 2008 were followed up for 6 months. The efficacy rate, complication rate, reintervention rate, and survival were analyzed. Results There were 61 self-expanding metal stents placed in 55 patients with malignant disease, with no intraoperative mortality. The immediate efficacy rate was 100% , the short-term( lt;30 days) efficacy rate was 94. 5% , and the survival rate in 6 months was 32. 7% . The complications included tumor ingrowth, excessive granulation tissue, stent migration, and restenosis. A total of 14 cases of complicationswere observed, in which two occurred during the short-term period ( lt; 30 days ) and the remaining complications occurred after 30 days. Conclusions Tracheobronchial stents can improve symptoms immediately for the patients with unresectable malignant central airway obstruction with fairly safety. The benefit of airway stents is particularly seen in the short-termperiod and the complications occur mainly after 30 days.
Abstract: ObjectiveTo explore the surgical characteristics of primary tracheal tumors treatment and its prognosis. Methods [WTBZ]We retrospectively investigated the clinical records of 38 patients with primary tracheal tumors in both Xiangya Hospital of Central South University and Hunan Provincial Tumor Hospital from Febuary 1982 to August 2009. There were 24 males and 14 females aged from 7 to 65 years. There were 2 benign lesions, 13 adenoid cystic carcinomas, 11 squamous cell carcinomas, 5 mucoepidermoid carcinomas, 4 adenocarcinomas and 3 other cell types. One patient with adenocarcinoma underwent exploratory thoracotomy only; 33 patients underwent tracheal resection and airway reconstruction, and according to the tumor growth characteristics, the surgeon applied circumferential tracheal resection with endtoend anostomosis or wedge resection; One patient had papilloma resected under fiber bronchoscopy; and three patients with locally advanced lesions underwent radiotherapy without surgery. The overall survival rate was calculated by the KaplanMeier method. The logrank method was used for comparing survival rates among different groups, characterized by cell types or surgical procedures. Results [WTBZ]The patient with exploratory thoracotomy died 3 days after surgery from respiratory failure. The perioperative mortality was 2.94% (1/34), and all the remaining 33 patients recovered and were discharged from hospital. Minor complications happened to 12 patients (35.29%), including 6 patients with pulmonary infection, 4 with atelectasis, and 2 with hoarseness. The followup time ranged from 6 months to 15 years. The followup rate was 97.29% (36/37). The threeunresected patients died within 6 months after hospital discharge. The 1, 5, and 10year survival rate for resected patients was88% （95%CI 0.77 to 0.99), 47%(95%CI 0.29 to 0.66), and 41% (95%CI 0.21 to 0.61), respectively. The survival rate of adenoid cystic carcinoma or mucoepidermoid carcinoma was significantly higher than that of squamous cell carcinoma or other tissue types (χ2=17.581, P=0.001). There was no statistical difference (χ2=0.021, P=0.886) in 5 year survival rate between wedge resection group at 63%(95%CI 0.34 to 0.91) and the segmental resection group at 77%(95%CI 0.44 to 0.99). Conclusions [WTBZ]Surgical treatment is safe and beneficial for primary tracheal tumors, and the pathological type is a significant prognostic factor after complete resection.
0bjective To investigate the prevalence of gastroesophageal reflux(GER)in acute exacerbation of chronic obstructive pulmonary disease(AECOPD)and to explore the influence of GER on the acidity of airway and pulmonary ventilation function in AECOPD patients．Methods Sixty-five hospitalized AECOPD patients and 20 healthy volunteers were enrolled in this study．According to “GER survey”questionnaire，AECOPD patients were allocated into GER group and non-GER group and expired breath condensate(EBC)pH and pulmonary ventilation function were detected．Breath condensate samples were collected using a specially designed condensing chamber(EcoScreen，Germany)．Results About 30.8％ (20／65)AECOPD patients were complicated with GER．EBC pH was lowest in GER group，middle in non-GER group and highest in the control group[6.oo±0.75 VS 6.76±0.77 and 7.97±0.12，both Plt;0.05)．The pulmonary ventilation function(FEV1、FEV1％pred)in AECOPD patients with GER were lower than those without GER(both Plt;0.05)．Conclusions There iS a high prevalence of GER in AECOPD patients．And the airway pH is reduced in AECOPD patients，especially when complicated with GER，which may play an important role in the development of COPD．
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 Respiratory syncytial virus ( RSV) is a primary cause of lower respiratory tract infections in children, and is also the cause for the development of asthma primarily in infants. However,the immunological mechanisms by which RSV enhances allergic sensitization and asthma remain unclear. The aimof this study was to examine the influence of RSV-infected airway epithelial cells on the activation and functions of rat myeloid dendritic cells ( mDCs) . Methods Rat airway epithelial cells ( RAECs) were infected by RSV. Then RSV-infected RAECs were co-cultured with rat mDCs, and the expression of cytokine and maturation markers on mDCs were examined by real time PCR and flow cytometry. To confirm this functional mDC maturation, allergenic mixed lymphocyte reaction ( MLR) were performed. Results Wefound that functional maturation of mDCs was induced by RSV-treated RAECs, as shown by their enhanced levels of OX40L and thymus- and activation-regulated chemokine ( TARC) mRNAs, which increased the expressions of major histocompatibility complex II ( MHCII) and CD86 costimulatorymolecules and promotedT-cell proliferation in mixed lymphocyte reactions. Conclusion Our results suggest that RSV-infected epithelial cells promote the maturation of mDCs that might support Th2 cell polarization and contribute to the pathogenesis of asthma.
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 examine the effects of Pseudomonas aeruginosa(PA)quorum-sensing systems on airway mucus hypersecretion．Methods Sixty Sprague-Dawley rats were intubated with a silicone tube pre-coated with PAO1(wild-type PA strain)，PAO1-JP2(quorum-sensing-mutant strain)or saline in the bronchus．After 28 days，the mRNA and protein expression of MUC5AC in the rats’bronchial epithelia were detected by RT-PCR，alcian blue／periodic acid—Schif(AB／PAS)staining and enzyme linked immunosorbent assay(ELISA)．Results In the PAO1 group，bronchiolar epithelium goblet cells by AB／PAS staining was significantly more than those in the PAO1-JP2 and control groups(both Plt;0．05)．The expression level of MUC5AC mRNA in the PAO1 group was significantly higher than those in the PAO1-JP2 and control groups(both P lt;0．05)．The ELISA showed that the concentration of MUC5AC protein in bronchoalveolar lavage fluid(BALF)in the PAO1 group was much higher than that in the PAO1-JP2 group(P lt;0．05)．Conclusion PA quorum-sensing system plays an important role in airway mucus hypersecretion
