[Levels of nerve growth factor and interleukin-4 in the induced sputum of children with cough variant asthma].

OBJECTIVE: To examine the levels of nerve growth factor (NGF) and interleukin-4 (IL-4) in the induced sputum of children with cough variant asthma (CVA), with the aim of studying the roles of NGF and IL-4 in childhood CVA. METHODS: Thirty-four children with CVA were enrolled in this study. Twenty healthy children were used as a normal control group. The induced sputum was separated into supernatant and cells. Hematoxylin and eosin staining was used to count differential cells. The expression of NGF and IL-4 in supernatant was measured using ELISA. The mRNA expression of NGF and IL-4 in cells was determined by Real-time PCR analysis. RESULTS: The percentage of eosinophils in the CVA group was significantly higher than in the control group [(13.4±3.6)% vs (2.6±1.7)%; P<0.01]. The expression of NGF and IL-4 protein and mRNA in induced sputum was significantly higher in the CVA group than in the control group (P<0.05). The expression of NGF and IL-4 protein and mRNA was positively correlated with the percentage of eosinophils (P<0.01). The expression of NGF and IL-4 protein and mRNA in induced sputum was significantly reduced in the CVA group after treatment (P<0.05). CONCLUSIONS: Eosinophils infiltration and increased expression of NGF and IL-4 play key roles in the development of childhood CVA, suggesting that they may be useful in the diagnosis and treatment of childhood CVA.

Functional roles of mannose-binding protein in the adhesion, cytotoxicity and phagocytosis of Acanthamoeba castellanii.

Acanthamoeba castellanii is a single-celled protozoan that is widely distributed in the environment and is a well-known of causing human keratitis, a vision-threatening infection. In this study, an ethyl methane sulfonate (EMS) and a selection of saccharide were applied to A. castellanii by chemical mutagenesis. To understand the functional roles of a mannose-binding protein (MBP). A. castellanii were treated with methyl-alpha-D-mannopyranoside abbreviated Man, with and without the EMS pre-treatment, and their adhesion and cytotoxicity were analyzed, using a human brain microvascular endothelial cell (HBMEC) as the target cell. Both EMS and Man mutants exhibited significantly decreased levels of MBP expression and cytotoxicity to HBMEC, but showed similar levels of binding to HBMEC, as compared with the wild type. Of interest was that the exogenous mannose inhibited amoebae (i.e., Man mutant) binding to the HBMEC by <20%. Only the mutant Man exhibited a significant decrease in bacterial uptake, as compared to the wild type, 0.020 vs 0.032 (p<0.05) and proteolytic activity. The results showed that MBP should be clearly provided as the pathogenic target candidate, to further target-based therapy, but EMS mutation should not be associated with initial adhesion and phagocytosis of A. castellanii.

[Effects of diesel exhaust particles inhalation on immediate reaction in asthma rats].

OBJECTIVE: The role of air pollution on asthma can not be ignored, diesel exhaust particles (DEP) in the air is one of the most important pollutants. This study aimed to investigate the effect and mechanism of DEP inhaled on immediate reaction in the asthma rats. METHOD: Sixty male Wistar rats of "Clean" grade, 6 - 7 week-old, with an average weight of (140 +/- 20) g were used in this study. The rats were randomly divided into 6 groups, 10 in each. Group A was treated with normal saline attack as a negative control, Group B with ovalbumin attack as a positive control. After ovalbumin attack, groups C, D, E, F continued to inhale DEP for 1 week, 2 weeks, 3 weeks and 4 weeks, respectively. The concentration of DEP was 200 microg/ml, the animals were subjected to inhalation of ultrasound nebulized DEP for 30 min per day. One week after all the attacks were concluded, Group A was stimulated with normal saline for 30 min, other groups were stimulated with ovalbumin. Then the airway resistance was determined with multi-channel signal acquisition and processing system and compared. The changes in neutrophils, eosinophils, and other inflammatory cells of BALF and the pathological changes in lung tissue, including epithelial cells loss, the inflammatory cells infiltration around the airway, basement membrane fibrosis, goblet cell hyperplasia etc. were observed. The concentration of IL-5 and gamma-interferon in the lung tissues, and the changes of serum IgE etc. were determined. RESULT: Airway resistance values of group A, B, C, D, E, F after ovalbumin excitation for 30 min were (3.56 +/- 0.21), (7.06 +/- 0.63), (6.46 +/- 0.38), (7.47 +/- 0.33), (8.87 +/- 0.61), (11.00 +/- 0.69) cm H2O/(ml.s). No airway hyperresponsiveness occurred in group A, while Groups B, C, D, E, F had higher airway resistance than group A, group E and F had higher airway resistance than that of group B, the differences were statistically significant. And the airway resistance was different in each group among 0 min, 10 min, 20 min and 30 min (F = 160.646, 148.901, 162.204, 156.186, P < 0.01 for both). The time of DEP inhalation and the airway resistance was positively correlated (r = 0.948, P < 0.01); IgE concentrations of the serum between groups B, C, D, E, F was not significantly different (P > 0.05), but higher than that of group A (F = 2.639, P < 0.01). The infiltrated inflammatory cells included eosinophils and lymphocytes, etc. The percentages of neutrophil(%) were (4.3 +/- 2.0), (9.7 +/- 5.2), (10.3 +/- 5.6), (13.0 +/- 5.2), (42.6 +/- 18.3), (55.3 +/- 6.9). The groups E and F had higher percentage than Group A and Group B (F = 114.226, P < 0.01). The percentages of eosinophils(%) were 0, (11.9 +/- 3.8), (15.8 +/- 6.3), (13.0 +/- 4.9), (21.1 +/- 5.6), (27.1 +/- 4.8). The difference between Groups B, C, D, E, F and Group A was statistically significant. There was significant difference between groups C, D, E, F and group B (F = 46.462, P < 0.05); Lung tissue biopsy in group A showed that the epithelial cells were intact, no inflammatory cells infiltrations were found around the airways, instead, mainly ciliated columnar epithelial cells and only a small number of goblet cells were seen without basement membrane fibrosis. With the inhalation of DEP, the epithelial cells showed gradual necrosis, disruption and loss, goblet cells showed hyperplasia, and infiltrations with inflammatory cells were seen around the airway. In the lung tissue, concentrations of IL-5 in group B, C, and E were (12.8 +/- 2.8), (17.1 +/- 5.2), (18.6 +/- 4.2) pg/mg, the difference between groups C, E and group B was statistically significant (F = 4.236, P < 0.01), the difference in gamma-interferon concentration among all groups was not statistically significance (F = 1.185, P > 0.05). CONCLUSION: DEP inhalation increased the airway responsiveness of asthma rats in immediate reaction, promoted the lung epithelial cell loss, inflammatory cell infiltration, basement membrane fibrosis and goblet cell hyperplasia.

Antagonism of Sophoricoside from Sophorica japonica on IL-5 / 천식및알레르기

BACKGROUND: Allergic disease is an inflammatory disease, whose main inflammatory cells are eosinophils, mast cells, and T lymphocytes. From that point, new therapeutic targets on allergic inflammation focusing on them are under investigation. We extracted four isoflavonoids from Sophorica japonica such as sophi, orobol, genistin, and genistein which has been known as PTK antagonist. We documented those three iso-flavonoids except genistein had an anti-inflammatory effect as potent as dexamethasone on carageen-induced ear model. Also they had antagonism on the Y-16 cell line, whose growth is dependent on IL-5. OBJECTIVES: From above results, in this experiment, we tried to find antagonistic effects of those compounds on IL-5 using the inhibition of eosinophil activation and survival in vitro and possibility of anti-allergic medicine. METHODS: LTC4 by RIA and ECP for degranulation by UniCAP(TM), which had been used previously were activation markers. RESULTS: Among those compounds, sophi was the most potent antagonist on IL-5 induced LTC4 release, degranulation, and survival. Orobol and genistin also had antagonism on them, but genistein, an antagonist of PTK didn't show any antagonistic effects. CONCLUSION: From these results, we concluded those three iso-flavonoids were IL-5 antagonist, and the mechanism of it might not be through PTK signaling.

O-glucogenistein inhibits eosinophil recruitment and nasal allergic symptoms in a murine model of nasal allergy / 천식및알레르기

BACKGROUND: Infiltration of eosinophils in the nasal mucosa is a consistent feature of nasal allergic inflammation. Various cytokines, especially interleukin-5(IL-5), were identified to play important roles in the infiltration and activation of eosinophils in nasal mucosa. Our previous study found that among 4 kinds of sophoricosides extracted from Sophora japonica, named sophi, orobol, genistin, and genistein, 3 compounds except genistein known as protein tyrosine kinase(PTK) inhibitor had anti-inflammatory and anti-IL-5 effects, and sophi was the most potent. OBJECTIVE: The goal of this study was to investigate the antagonism of sophi on the nasal eosinophilia in ovalbumin(OA)-sensitized murine nasal allergy model. METHODS: Male BALB/c mice sensitized intraperitoneally and then topically with OA were treated with sophi(10 or 30mg/kg) or anti-mouse IL-5 monoclonal antibody(anti-IL-5 mAb, 1mg/Kg) intravenously 1 hour before challenge. The effect of sophi on the infiltration of eosinophils into the nasal mucosa, peripheral blood eosinophilia, nasal symptom, and OA-specific IgE antibody production were evaluated. Results: Administration of sophi(10, 30mg/kg) significantly inhibited the nasal eosinophil infiltration and nasal symptom compared to that of anti-IL-5 mAb. But eosinophil count inthe peripheral blood and the titer of OA-specific IgE were not affected by sophi. CONCLUSION: Sophi inhibited not only the tissue eosinophilia but also the acute nasal allergic symptom. These findings suggest that sophi has anti-eosinophilic cytokine activity and also plays blockade of early allergic reaction. Taken together, sophi may be a candidate for new anti-allergic medicine.