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 investigate the effects of diesel exhaust particles ( DEP) on the production of CCL11, CCL24 and CCL26 in asthmatic rats. Methods Fifty SD rats were randomly divided into five groups. Group A was an normal control group. The rats in group B, C, D, and E were sensitized and challenged by ovalbumin ( OVA) to establish asthma model. Then the rats in the group C, D, E were inhaled DEP for 1, 2, 3 weeks, respectively. Lung tissue and brouchoalveolar lavage fluid ( BALF) were collected for detection of CCL11, CCL24, and CCL26 expression by ELISA and q-RT-PCR. Results The transcription of CCL 24, CCL26 gene and the production of CCL24 and CCL26 protein increased significantly compared with the control group ( P lt;0. 05) , and were positively associated with the DEP inhalation time. However, CCL11 gene and protein expression were not changed significantly compared with the control. Conclusion The exposure to DEP can induce the production of CCL24 and CCL26 in the asthmaic rats, which might aggravateairway hyperresponsiveness.
ObjectiveTo determine whether there is an imbalance of KLF2/RelA in peripheral blood neutrophils in patients with bronchial asthma, and explore the relationship between KLF2/RelA imbalance and neutrophil apoptosis.MethodsFrom April 2011 to April 2012, a total of 39 patients with acute attack of asthma in Hunan People's Hospital and Third People's Hospital of Changsha were enrolled, with 13 cases in mild asthma group, 17 cases in moderate asthma group, and 9 cases in severe asthma group. Fifteen healthy subjects were recruited as control group. Peripheral blood were collected from all subjects followed by separation of neutrophils. The apoptosis of neutrophils were measured by flow cytometry. The expression of KLF2 and RelA were detected by Western blot. The relationship between the ratio of KLF2/RelA and neutrophil apoptosis rate was analyzed by Pearson correlation test.ResultsNeutrophil apoptosis rates in the mild, moderate and severe asthma groups [(4.45±0.76)%, (2.10±0.25)%, (1.81±0.67)%, repectively] were lower than that in the healthy control group [(5.36±0.57)%, all P<0.01]. The apoptosis rates of neutrophils in the moderate and severe asthma groups were lower than that in the mild asthma group (bothP<0.01), but there was no significant difference between the moderate asthma group and the severe asthma group (P>0.05). The ratios of neutrophil KLF2/RelA in the mild, moderate and severe asthma groups were lower than that in the normal control group (0.667±0.351, 0.384±0.203, 0.536±0.293vs. 4.038±2.011, all P<0.01). There was no significant difference between the groups of mild, moderate and severe asthma (P>0.05). The neutrophil apoptosis rate was positively correlated with the percentage of neutrophil KLF2/RelA (r=0.592 0, P<0.000 1).ConclusionThere is an imbalance of KLF2/RelA in peripheral blood neutrophils in patients with bronchial asthma, and the imbalance of KLF2/RelA may be the mechanism of apoptosis of peripheral blood neutrophils.
【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 review literatures regarding the diagnosis of asthma with the measurement of exhaled nitric oxide( eNO) and assess the effectiveness and accuracy of eNO in the diagnosis of asthma.Methods MEDLINE, OVID, CBMdisc, CNKI( 1991 to 2008) for studies involving the diagnostic value of eNO were searched, and references of included studies were also hand searched. QUADAS ( Quality Assessment of Diagnostic Accuracy Studies) items were used for quality assessment in the systematic review. Meta-disc software was used to analyze heterogeneity. Sensitivity, specificity and summary diagnostic odds ratio( SDOR) were used for the pooled analysis. The summary receiver operating characteristic ( SROC)curves were drew and the summary areas under the SROC ( SAUC) were calculated. Finally, sensitivity analysis was performed. Results Eleven literatures with15 studies were included. These 15 studies had well controlled the bias of partial verification, differential verification, incorporation and withdrawals. The possibility of the disease progression bias was less and the reference standard review could have a greater bias. The spectrumcomposition of a study, the inclusion and exclusion criteria and the reporting quality were poorly reported. In statistical analysis, the totally pooled sensitivity, pooled specificity, SDOR, SAUC of the measurement of eNO in the diagnosis of asthma was 0. 68, 0. 79, 12. 73, 0. 8446, respectively. Sensitivity analysis demonstrated no disproportionate influences of individual study. Conclusions eNO has a certain value in the diagnosis of asthma. To make further analysis, more studies with high quality are needed.
目前哮喘的治療已經取得了顯著的進步, 大多數哮喘按全球哮喘防治創議( GINA) 推薦的治療方案進行規范治療都可以獲得良好的臨床控制, 但仍有部分中重度患者難以獲得良好的臨床控制, 所以仍有必要探索針對這一部分哮喘人群的更好的干預措施。
Objective To investigate the effects of 1, 25-( OH) 2D3 on the expression of matrix metalloprotease-9 ( MMP-9) and nuclear factor κB ( NF-κB) activity in a murine model of chronic asthma. Methods BALB/ c mice were sensitized and challenged with ovalbumin to establish chronic asthmatic model. The animals were randomly divided into a control group, an asthma group and a VD group. Lung sections from the mice were stained by HE and Masson’s trichrome, respectively. Morphometric analysis of the stained sections was performed using computerized image analysis system. Nuclear translocation of NF-κB p65 was examined using Western blot. The level of IκBαwas detected with real-time quantitative PCR ( RTPCR) and Western blot. In addition, the expression of MMP-9 in both activity and mRNA level was detected by gelatin zymograph and RT-PCR, respectively. Results Prominent airway remodeling developed in the asthma group, including the inflammatory cell infiltration, subepithelial collagen deposition and increased airway smooth muscle mass. In contrast, 1, 25-( OH) 2D3 attenuated these established structural changes of the airways. Stimulation with OVA induced a 7. 87-fold increase in the MMP-9 activity compared with that in the control group, and 1, 25-( OH) 2D3 treatment only induced a 3. 46-fold increase in the MMP-9 activity compared with that in the control group ( P lt;0. 05) . The mRNA level of MMP-9 in the VD group ( 3.16 ± 0.09) was decreased compared with the asthma group ( 5.74 ±0.13) ( P lt;0.05) , but itwas still higher than that in the control group ( 0.57 ±0.08) ( P lt;0.05) . 1, 25-( OH) 2D3 reduced the nuclear translocation of NF-κB p65 while up-regulated the IκBα level in lung tissue of chronic asthma. Conclusions 1, 25- ( OH) 2D3 can inhibit the NF-κB activity and down-regulate the expression of MMP-9 in lung tissue of chronic asthma, thus alleviating the established chronic asthma-induced airway remodeling.
Objective To investigate levels of 8-isoprostane in serum of patients with bronchial asthma. Methods Eighteen patients diagnosed with acute exacerbation of asthma were enrolled as the experimental group from Department of Respiratory Medicine from February 2009 to August 2009. After treatment all the patients reached remission. Twenty healthy workers from Department of Respiratory Medicine were enrolled as the control group in August 2009. The levels of 8-isoprostane in serum of all subjects were measured, and their FEV1% pred was also evaluated. Results The levels of 8-isoprostane in serum were significantly higher in patients with acute exacerbation of asthma compared with those in remission stage and the healthy control group [ ( 157. 46 ±46. 99) pg/mL vs. ( 43. 52 ±13. 62) pg/mL and( 15. 23 ±1. 96) pg/mL, P lt;0. 01] . Meanwhile the levels of 8-isoprostane in serum of patients with asthma in remission stage were significantly higher compared with the healthy control group ( P lt;0. 05) . The levels of 8-isoprostane in serum were negatively correlated with FEV1% pred in the asthma group( r = - 0. 533,P lt;0. 05) . Conclusions 8-isoprostane as amarker of oxidative stress response involves in the pathogenesis of asthma. Monitoring 8-isoprostane levels in serum may reflect the state of oxidative stress, and may be useful for severity judgment and follow-up of treatment effectiveness in patients with asthma.
ObjectiveTo investigate the drug use,symptoms and lung function of asthma patients in communities. MethodsA cooperation project of "Cohort Study in Putuo District Health Survey" was conducted by Shanghai Putuo People's Hospital and the Harvard School of Public Health from August 2007 to January 2010 .Home questionnaires were collected in the communities in 13 residential quarters in Changzheng town,Ganquan town in Putuo District,Shanghai. ResultsA total of 27 042 respondents was inquired,in which there were a total of 488 asthma patients with the total prevalence rate of 1.80 percent. 189 cases (47.49%) frequently used anti-asthma drugs in nearly a year,and 209 cases (52.51%) less frequently used anti-asthma drugs. In the patients frequently using anti-asthma drugs,FEV1%pred was (65.30±25.78)% with FEV1%pred ≥80% in 50 cases (32.68%).St. George score was 36.80±28.02 in this subgroup.More symptoms presented in 129 cases(69.73%)in the past year. In the patients less frequently using anti-asthma drugs,FEV1%pred was (81.55±19.01)% with FEV1%pred ≥80% in 99 cases (56.57%). St. George score was 7.61±15.56.There were symptoms presented in 42 cases (20.59%) in the past year. ConclusionMore than half asthmatic patients do not receive regular medicine treatment in communities in Shanghai. Some patients still suffer from asthma symptoms long with poor St. George score and lung function although frequently using anti-asthma medication,suggesting asthma treatment is not standard.
Objective To investigate the effects of dust mite allergen Derp1 on the expressions of IL-6 and IL-8 in primary rat bronchial epithelial cells. Methods The primary rat bronchial epithelial cells were divided into a control group and three experimental groups. In the experimental groups, the cells were cultured with 3 different concentrations of Derp1 ( 1, 5, 10 μg/mL) for 3 different time ( 4, 8, 24 h) .Inverted microscope was employed to observe the morphological changes of bronchial epithelial cells and intercellular space, and supernatants were assayed for IL-6 and IL-8 with ELISA. Results Complete flattening of single cells layer was observed in the control group. In the experimental groups, the cells treated with Derp1 allergen showed no obvious changes in the cell morphology and intercellular space. However,There was a significant change in the level of cytokines production compared with the control group. The levels of IL-6 and IL-8 began to rise at 4 h, and reach to high level at 8 h, especially in the 5 and 10 μg/mL groups ( P lt;0. 01) . In the 24h group, the concentrations further increased but not reach statistical difference compared with 8h group ( P gt; 0. 05) . Conclusions The Derp1 allergen can stimulate the release of inflammatory cytokines such as IL-6 and IL-8 fromthe rat trachea- bronchia epithelial cells. It is suggested that dust mite allergen -induced cytokines may play important roles in the pathogenesis of allergic asthma.