OBJECTIVETo investigate the effect of chloroquine on airway hyperresponsiveness in asthmatic mice and explore the possible mechanism.

METHODSBalb/c mouse models of asthma established using OVA received intraperitoneal injections of chloroquine, dexamethasone, or both prior to OVA challenge. Within 24 h after the final challenge, airway hyper- responsiveness (AHR) of the mice was assessed, and the total cell count and the counts of different cell populations in the bronchoalveolar lavage fluid (BALF) were determined under light microscopy. The severity of lung inflammation was evaluated using HE staining, and the concentrations of IL-6 and PGF2α in the BALF were detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSChloroquine pretreatment significantly decreased AHR (P<0.001) in the asthmatic mice and reduced the total cell count (P<0.01), eosinophils (P<0.001), neutrophils (P<0.01), and PGF2α levels in the BALF. Chloroquine combined with low-dose dexamethasone significantly lessened inflammations around the bronchioles (P<0.05) and blood vessels (P<0.01) in the lung tissue, and obviously lowered IL-6 (P<0.05) and PGF2α (P<0.001) in the BALF in the asthmatic mice.

CONCLUSIONChloroquine can inhibit AHR in asthmatic mice and produce better anti-inflammatory effect when combined with dexamethasone for treatment of neutrophilic asthma.

Animals ; Asthma ; chemically induced ; drug therapy ; Bronchoalveolar Lavage Fluid ; cytology ; Chloroquine ; pharmacology ; Dexamethasone ; pharmacology ; Dinoprost ; metabolism ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; cytology ; Inflammation ; pathology ; Interleukin-6 ; metabolism ; Leukocyte Count ; Lung ; pathology ; Mice ; Mice, Inbred BALB C ; Neutrophils ; cytology

OBJECTIVE: To study the role of P-JNK and P-c-Jun of JNK (c-Jun N-terminal kinase) on nasal mucosa remodeling in allergic rhinitis rats. METHOD: Sixty male Wistar rats (weighing about 200-250 g) were randomly divided into AR group (A group) and B group(control group). The rats in A group were sensitized for inducing AR by intraperitoneal injection of ovalbumin and Al(OH)₃. Rats in group A were randomized into A4, A8 and A12 group (each had 10 rats). Ovalbumin was dropped in each nasal cavity of every rat for 4,8,12 weeks, respectively. Rats in group B were sensitized by saline instead of OVA, and were also divided into B4, B8 and B12 group. Each group had 10 rats. Pathological changes of nasal mucosa in each period were observed by hematoxylin and eosin stain dyeing. The phosphorylation of JNK and c-Jun were tested by immunohistochemistry. RESULT: In A8 group, mucosal congestion and edema thickening with inflammatory cells infiltration of eosinophils were observed in the eighth week, and the inflammatory changes were significantly increased as time went on. The mean absorbance values of P-JNK and P-c-Jun in A group were significantly higher than those in the corresponding B group (all P < 0.01). Moreover, the mean absorbance values of A12 group were significantly higher than A4 group and A8 group (all P < 0.01 ). CONCLUSION The expression of P-JNK and P-c-Jun in the process of nasal mucosa remodeling in allergic rhinitis rats were increased, which suggested that P-JNK and P-c-Jun played important roles in nasal mucosa remodeling of the allergic rhinitis rats.
Airway Remodeling ; Animals ; Disease Models, Animal ; Eosinophils ; cytology ; Injections, Intraperitoneal ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Male ; Nasal Mucosa ; metabolism ; Ovalbumin ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Rhinitis, Allergic ; metabolism ; Signal Transduction

OBJECTIVE: To discuss the effect of different doses intranasal corticosteroids on remodeling of allergic rhinitis (AR) mice nasal mucosa and expression level of matrix metalloproteinase-9 (MMP-9). METHOD: Thirty BALB/c female mice were divided into five groups randomly and received OVA or normal saline (NS) with intraperitoneal injection or nasal challenge, respectively. The treatment groups received additional different doses of budesonide (0.6 μg/20 g, 3.0 μg/20 g and 15.0 μg/20 g) daily for 16 weeks. We assessed the nasal symptoms at 4 and 16 weeks. Collected the mice nasal tissue, and then stained with hematoxylin-eosin, Masson's Trichrome, and periodic acid-schiff respectively to evaluate airway remodeling at 16 weeks. MMP-9 was measured with enzyme-linked immunosorbent assay (ELISA). Result: Times of rubbing, sneezes and infiltrate of eosinophil increased more in B group than in A group, and subepithelial fibrosis, collagen deposition, goblet cell hyperplasia, and submucosal gland hypertrophy were only observed in B group at 16 weeks. The nasal symptoms and eosinophil infiltration were inhibited by treatment with budesonide from a dose of 0.6 μg onwards, while the prevention of structure changes was only observed with 3.0 μg onwards. In addition, intranasal budesonide reduced MMP-9 in the nasal of AR mice. CONCLUSION The study suggests that higher dose intranasal corticosteroids might inhibit the airway remodeling of nasal mucosa by reducing MMP-9.
Airway Remodeling ; Animals ; Budesonide ; pharmacology ; Disease Models, Animal ; Eosinophils ; cytology ; Female ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa ; drug effects ; Rhinitis, Allergic ; drug therapy ; metabolism

OBJECTIVE: To study the expression of interleukin-17 (IL-17) in nasal polyps from both atopic and nonatopic patients, and its associations with histological features of polyps tissue. METHOD: Thirty patients with nasal polyps (NP) were included and divided into atopic and nonatopic groups according to the skin prick test. Histological characteristics were assessed by eosinophilic infiltration with HE staining. IL-17 expression in polyps tissue was detected by ELISA and RT-PCR. RESULT: Eosinophilic infiltration was significantly higher in atopic NP patients than in nonatopic NP patients (P < 0.01). IL-17 protein and IL-17 mRNA levels were significantly upregulated in both atopic (P < 0.01) and nonatopic (P < 0.05) patients compared with controls. Furthermore, IL-17 levels were significantly higher in the atopic group than in nonatopic group. Significantly positive correlations were found between IL-17 levels and eosinophilic infiltration in NP patients. CONCLUSION These results indicated that expression of IL-17 was significantly upregulated in NP patients and was especially higher in atopic NP patients, suggesting that IL-17 may play an important role in the pathogenesis of NP and atopy may contribute to NP by stimulating the production of IL-17.
Adult ; Aged ; Eosinophils ; cytology ; Female ; Humans ; Hypersensitivity, Immediate ; metabolism ; pathology ; Interleukin-17 ; metabolism ; Male ; Middle Aged ; Nasal Polyps ; metabolism ; pathology ; Young Adult

Phytoestrogens, a group of plant-derived non-steroidal compounds that can behave as estrogens by binding to estrogen receptors, have drawn great attention for their potentially beneficial effects on human health. However, there are few studies investigating the potential side effects of phytoestrogens on the reproductive system. The present study was to elucidate the effects of 17β-estradiol (E2), progesterone (P4), and phytoestrogens genistein (Gen), resveratrol (Res), and phloretin (Phl) on eosinophilic infiltration of the ovariectomized rat uterus and endometrial vascular permeability, and to analyze the underlying mechanisms. The ovariectomized rats received daily subcutaneous injections of E2, E2+P4, P4, Gen, Res, Phl, or an equivalent volume of vehicle for 21 days, and sham-operated animals (Sham rats) were used as the controls. Hematoxylin-eosin staining revealed a marked increase in uterine eosinophilic infiltrations in ovariectomized rats treated with E2, E2+P4 or P4, which was associated with increased expression of vascular endothelial growth factor (VEGF), nuclear factor-κB (NF-κB), and tumor necrosis factor-α (TNF-α) proteins as determined by immunohistochemical and Western blot analysis. However, all three phytoestrogens had no markedly effect on the uterine eosinophilic infiltration and the expressions of VEGF, NF-κB, and TNF-α in the uterus of ovariectomized rats. Our data demonstrate that E2 alone or in combination with P4 increases uterine eosinophilic infiltration which is related with vascular hyperpermeability caused by VEGF, NF-κB and TNF-α, whereas phytoestrogens Gen, Res, and Phl, have no such an effect.
Animals ; Endothelium, Vascular ; drug effects ; Eosinophils ; cytology ; Estradiol ; pharmacology ; Estrogens ; pharmacology ; Female ; Genistein ; pharmacology ; NF-kappa B ; metabolism ; Ovariectomy ; Permeability ; Phloretin ; pharmacology ; Phytoestrogens ; pharmacology ; Progesterone ; pharmacology ; Rats ; Stilbenes ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism ; Uterus ; drug effects ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVEIt is known that Siglec-8 is selectively expressed on human eosinophils at a high level and mediates eosinophil apoptosis when crosslinked with its antibody. The aim of our review is to elucidate the molecular and biological characteristic of Siglec-8 and then discuss the function and possible mechanisms of Siglec-8 in eosinophils. Thereby, we will expand our understanding to the regulation of eosinophil apoptosis, and provide important clues to the treatment of asthma and other hyper-eosinophilic diseases.

DATA SOURCESMost articles were identified by searching of PubMed online resources using the key term Siglecs.

STUDY SELECTIONMainly original milestone articles and critical reviews written by major pioneer investigators in the field were selected.

RESULTSSiglec-8 is selectively expressed on human eosinophil and can specifically induce eosinophil apoptosis.

CONCLUSIONThe restricted expression of Siglec-8 on human eosinophil and the rapid progress in understanding its role as cell signaling and activation of death receptors have made it an attractive target for treatment of asthma and other hyper-eosinophilic diseases.

Apoptosis ; genetics ; physiology ; Asthma ; metabolism ; therapy ; Eosinophils ; cytology ; metabolism ; Humans ; Sialic Acid Binding Immunoglobulin-like Lectins ; metabolism

OBJECTIVETo establish the method of cytological examination and the normal reference values for hypertonic saline solution-induced sputum of healthy children (age range from 5 to 15 years) with physical examination in Guangzhou.

METHODA total of 352 children, 5 to 15 years old, were enrolled from primary school and middle school in Guangzhou from January to December, 2010. All subjects completed a standardized questionnaire on the presence of respiratory, allergic symptoms and family history, the medical history and the physical examination was performed by doctors, lung function (forced expiratory volume at 1 s in predicted normal, FEV(1)%) was determined. There were 266 healthy children (137 males, 129 females) who were selected and undergone hypertonic saline solution induction of sputum, and cytological examination was performed. Hypertonic saline (5%) was nebulized and inhaled for 15 - 30 min. No expectoration within 30 min was defined as failure, and the procedure was terminated. The part of opaque and higher density sputum samples was detected by cytology. The proportion of neutrophils, lymphocytes, eosinophils, macrophages and monocytes was calculated. This study was approved by the institutional Ethics Review Committee of First Affiliated Hospital of Guangzhou Medical College. Informed consent was obtained from the legal guardians of all participants following a detailed description of the purpose and potential benefits of the study.

RESULTThere were 175 subjects' induced sputum specimens (175/266, 65.8%), non-qualified sputum samples were obtained from 16 of the subjects. The proportions of median (IQR) of lymphocytes were 0.012 (0.020), 95%CI were ranged from 0.015 to 0.022; neutrophils 0.207 (0.330), 95%CI 0.266 - 0.356 macrophages 0.761 (0.327), 95%CI 0.607 - 0.699; eosinophils 0.004 (0.019), 95%CI 0.013 - 0.022. There were no significant differences in proportions of cytological findings of female or male, different age groups and second-hand smoking or not (all P > 0.05). The incidence of adverse event was 4.40% (7/159).

CONCLUSIONThe method and the preliminary data may be used for research, diagnosis and treatment of patients with chronic cough and airway inflammation.

Adolescent ; Child ; Child, Preschool ; China ; Cough ; diagnosis ; physiopathology ; Eosinophils ; cytology ; Female ; Forced Expiratory Volume ; Humans ; Leukocyte Count ; Lymphocyte Count ; Lymphocytes ; cytology ; Male ; Monocytes ; cytology ; Neutrophils ; cytology ; Reference Values ; Saline Solution, Hypertonic ; chemistry ; Sputum ; cytology ; metabolism

DNA methylation may regulate gene expression by restricting the access of transcription factors. We have previously demonstrated that GATA-1 regulates the transcription of the CCR3 gene by dynamically interacting with both positively and negatively acting GATA elements of high affinity binding in the proximal promoter region including exon 1. Exon 1 has three CpG sites, two of which are positioned at the negatively acting GATA elements. We hypothesized that the methylation of these two CpGs sites might preclude GATA-1 binding to the negatively acting GATA elements and, as a result, increase the availability of GATA-1 to the positively acting GATA element, thereby contributing to an increase in GATA-1-mediated transcription of the gene. To this end, we determined the methylation of the three CpG sites by bisulfate pyrosequencing in peripheral blood eosinophils, cord blood (CB)-derived eosinophils, PBMCs, and cell lines that vary in CCR3 mRNA expression. Our results demonstrated that methylation of CpG sites at the negatively acting GATA elements severely reduced GATA-1 binding and augmented transcription activity in vitro. In agreement, methylation of these CpG sites positively correlated with CCR3 mRNA expression in the primary cells and cell lines examined. Interestingly, methylation patterns of these three CpG sites in CB-derived eosinophils mostly resembled those in peripheral blood eosinophils. These results suggest that methylation of CpG sites at the GATA elements in the regulatory regions fine-tunes CCR3 transcription.
Binding Sites ; Cell Line ; *CpG Islands ; DNA Methylation ; Enhancer Elements, Genetic ; Eosinophils/cytology/*metabolism ; Exons ; Fetal Blood/cytology/metabolism ; GATA1 Transcription Factor/*genetics/metabolism ; Gene Expression Regulation ; Humans ; Promoter Regions, Genetic ; RNA, Messenger/metabolism ; Receptors, CCR3/*genetics/metabolism ; Sequence Analysis, DNA ; *Transcription, Genetic

To determine the impact of IL-23 knockdown by RNA interference on the development and severity of ovalbumin (OVA)-induced asthmatic inflammation, and the potential mechanisms in mice, the IL-23-specific RNAi-expressing pSRZsi-IL-23p19 plasmid was constructed and inhaled into OVA-sensitized mice before each challenge, as compared with that of control mice treated with alum or budesonide. Inhalation of the pSRZsi-IL-23p19, significantly reduced the levels of OVA-challenge induced IL-23 in the lung tissues by nearly 75%, determined by RT-PCR. In addition, knockdown of IL-23 expression dramatically reduced the numbers of eosinophils and neutrophils in BALF and mitigated inflammation in the lungs of asthmatic mice. Furthermore, knockdown of IL-23 expression significantly decreased the levels of serum IgE, IL-23, IL-17, and IL-4, but not IFNgamma, and its anti-inflammatory effects were similar to or better than that of treatment with budesonide in asthmatic mice. Our data support the notion that IL-23 and associated Th17 responses contribute to the pathogenic process of bronchial asthma. Knockdown of IL-23 by RNAi effectively inhibits asthmatic inflammation, which is associated with mitigating the production of IL-17 and IL-4 in asthmatic mice.
Animals ; Asthma/chemically induced/genetics/metabolism/*prevention & control ; Bronchoalveolar Lavage Fluid/cytology ; Enzyme-Linked Immunosorbent Assay ; Eosinophils ; Female ; Inflammation/metabolism ; Interleukin-23/*genetics ; Leukocyte Count ; Mice ; Mice, Inbred BALB C ; Neutrophils ; Ovalbumin/pharmacology ; Plasmids/genetics ; *RNA Interference ; RNA, Small Interfering/*genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Th17 Cells/immunology

OBJECTIVE: To explore the expression of T-bet/GATA-3 in nasal mucosa tissue of allergic rhinitis rat and to investigate the association between the expression of T-bet/GATA-3 and the eosinophil count. METHOD: Twenty SD rats were randomly divided into a control group and an allergic rhinitis group. The allergic rhinitis rat model was induced with ovalbumin. The total eosinophils were counted in the nasal mucosa. The concentrations of IL-4, IL-5 and IFN-gamma in nasal lavage fluid were measured by ELISA. The mRNA and protein expressions of IL-4, IL-5, IFN-gamma, T-bet and GATA-3 in the nasal mucosa were detected by RT-PCR and Western blot respectively. RESULT: The main inflammatory cells were eosinophils in the nasal mucosa of allergic rhinitis rats. The level of IL-4, IL-5 and IFN-gamma in control group was significantly higher than that in allergic rhinitis group (P < 0.01). The mRNA and protein expression of IFN-gamma and T-bet in allergic rhinitis group was significantly higher than that in control group (P < 0.01). While the mRNA and protein expression of IL-4, IL-5 and GATA-3 in control group was significantly higher than that in allergic rhinitis group (P < 0.01). The ratio of protein expression of T-bet and GATA-3 was negatively correlated with the eosinophil count, IL-4 and IL-5, but positively with the concentrations of IFN-gamma. CONCLUSION The imbalance of transcription factor GATA-3 and T-bet has a close correlation with the eosinophil count, and may play a key role in the formation of allergic rhinitis.
Animals ; Cell Count ; Eosinophils ; cytology ; Female ; GATA3 Transcription Factor ; metabolism ; Hypersensitivity ; immunology ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Rhinitis ; immunology ; metabolism ; T-Box Domain Proteins ; metabolism ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism