IL-17C expression in nasal epithelial cells of chronic rhinosinusitis with nasal polyposis

Interleukin 17C (IL-17C) is a functionally distinct member of the IL-17 family that is selectively induced in epithelia by bacterial challenge and inflammatory stimuli. The goal of this study was to explore the expression of IL-17C in nasal epithelial cells and their role in the pathogenesis of chronic rhinosinusitis with nasal polyposis (CRSwNPs). IL-17C expression was detected using immunohistochemistry (IHC) of the epithelial cell layers and using the western blot assay on whole tissue homogenates from control subjects (n = 10) and CRSwNP patients [10 non-eosinophilic polyps and 10 eosinophilic polyps (EPs)]. Expression of IL-17C and P47-phox were evaluated in the human nasal epithelial cells (RPMI-2650 cells) after treatment with staphylococcal enterotoxin B (SEB) and pretreatment with reactive oxygen species (ROS) scavenger, N-acetyl l-cysteine (NAC). Finally, IL-17C expression was demonstrated in eosinophilic rhinosinusitis murine model using IHC. Epithelial expression of IL-17C was higher in nasal polyps (especially in EPs) compared to control mucosa. SEB increased the expression of IL-17C and P47-phox in RPMI-2650 cells. SEB-induced expressions of both IL-17C and P47-phox were significantly decreased in NAC-pretreated cells. Epithelial expression of IL-17C was significantly higher in experimental mice compared to control mice. SEB-induced IL-17C expression in nasal epithelial cells is mediated by ROS production. This pathway may be associated with the pathogenesis of CRSwNP, especially eosinophilic nasal polyps.     

Expression of the cysteinyl leukotriene 1 receptor and glucocorticoid receptor-β in nasal polyps

The objective of this study was to analyze the expression of cysteinyl leukotriene 1 (CysLT1) receptor and glucocorticoid receptors (GRs) in nasal polyps, and to evaluate the relationship between the expression of CysLT1 receptors and that of GRs. Nasal polyps were taken from 32 patients of chronic rhinosinusitis with nasal polyposis. Samples of middle turbinate from seven healthy subjects were used as controls. Specimens were immunohistochemically stained for CysLT1 receptor, GR-α and GR-β receptor, and were quantified in the unit area of the tissues. Numbers of CysLT1 receptor-positive cells were much increased in nasal polyps than in middle turbinate (281 ± 67 vs. 157 ± 85 cells/mm², P = .01). There was no significant difference in the numbers of GR-α positive cells between nasal polyps and normal turbinate mucosa. GR-β positive cells were increased in nasal polyps as compared to normal turbinate mucosa (36 ± 8 vs. 19 ± 7 cells/mm², P = .03). A significant relationship was found between the expression of CysLT1 receptor and GR-β in nasal polyps (R = .525, P = .04), whereas there was no significant relationship between the expression of CysLT1 receptor and GR-α in nasal polyps. Our study shows that CysLT1 receptor expression predominates on GR-β over-expressed polyps. This may suggest the additional effect of CysLT1 receptor antagonist for the treatment of nasal polyposis resistant to steroid alone.     

Eosinophils and mast cells: a comparison of nasal mucosa histology and cytology to markers in nasal discharge in patients with chronic sino-nasal diseases

Allergic rhinitis (AR), nasal polyps (NP) as well as chronic rhinosinusitis (CRS) are all known to be associated with eosinophilic infiltration and elevated numbers of mast cells (MC) within the mucosa. Both cell types and their markers eosinophilic cationic protein (ECP) and tryptase are utilized in the diagnosis and management of chronic sino-nasal diseases. Mucosal cytology samples were gathered by cytobrush, histological samples were obtained from the inferior turbinate. In both sample sets, the number of eosinophils and MC was determined. Their corresponding markers ECP and tryptase were quantified from nasal discharge. Patients were grouped with reference to their main diagnosis: AR (n = 34), NP (n = 25), CRS (n = 27) and controls (n = 34). Eosinophil counts from cytobrush and ECP levels were significantly elevated in NP compared to all other groups—31- and 13-fold over control, respectively. However, histologic review did not reveal any difference in eosinophil count among groups. Tryptase was significantly elevated threefold in AR versus CRS and controls. No correlation to cytological and histological MC counts could be found. ECP levels in nasal discharge as well as eosinophil counts can provide useful information with regard to the diagnosis. Likewise, tryptase concentrations can do. The presented data show that the measurement of markers in nasal discharge is superior in differentiating among diagnosis groups. Given that the collection of nasal secretions is more comfortable for patients than the more invasive techniques, we recommend first line ECP and tryptase testing performed on nasal secretions.     

金黄色葡萄球菌肠毒素B对人鼻黏膜上皮细胞的促炎作用

目的探讨金黄色葡萄球菌肠毒素B（staphylococcus aureus enterotoxin B，SEB）对原代培养的人鼻黏膜上皮细胞释放促炎因子和（或）趋化因子的作用。方法将无血清原代培养的人鼻息肉及下鼻甲上皮细胞分别在SEB1、10、100ns／ml，白细胞介素（intedeukin，IL）1β 20ng／ml及SEB 10ng／ml＋地塞米松13ng／ml等不同条件下孵育，12、24和48h后采用原位杂交方法检测鼻黏膜上皮细胞分泌的IL-5和粒细胞巨噬细胞集落刺激因子（granuloyte macrophagc colony stimulating factor，GM-CSF）在mRNA水平的变化。结果①SEB刺激鼻黏膜上皮时，IL-5和GM—CSF mRNA表达增加，在一定范围内呈现时间和剂量依赖性，以10ng／ml、24h时最明显（P〈0．05），且鼻息肉组的表达高于下鼻甲组，差异具有统计学意义（P〈0．05）；②IL-1β 20ng／ml组IL-5和GM-CSF mRNA表达增加不及SEB 10ng／ml组明显，差异具有统计学意义（P值均〈0．05）；③SEB 10ng/ml＋地塞米松13ng／ml组的IL-5和GM-CSF mRNA表达强度较SEB 10ng／ml组弱，差异具有统计学意义（P值均〈0．05）。结论SEB对原代培养的人鼻黏膜上皮细胞具有促炎作用。     

Interleukin-17A expression in patients presenting with nasal polyposis

Sinonasal polyposis (SNP) is a chronic inflammatory pathology of the nasal/paranasal cavities which affects from 1%-4% of the population. Although polyps seem to be a manifestation of chronic inflammation of nasal/paranasal sinus mucosa in both allergic and non-allergic subjects, the pathogenesis of nasal polyposis remains unknown. Interleukin-17A (IL-17A) is a key inflammatory cytokine in many disorders. Little attention has been paid to the role of IL-17A in chronic inflammatory disorders.ObjectiveTo investigate the expression of IL-17A in the SNP and verify if this expression is a marker of good or bad prognosis.MethodProspective study with 25 patients presenting with SNP were subjected to the immunohistochemistry technique. After a skin prick test, all patients were divided into atopic and nonatopic groups, and asthmatic or non-asthmatic.ResultsThe IL-17A expression was observed in both atopic and nonatopic patients. The numbers of IL-17A positive cells were greater in nasal polyps of atopic patients than nonatopic (p = 0.0128).ConclusionThese results indicate that IL-17A may play an important role in the pathology of SNP. Considering the inflammatory properties of IL-17A, this study suggests that it could increase susceptibility to atopy and asthma.     

Effects of topical nasal steroids and diclofenac on the nasal mucosa during hyperbaric oxygen therapy: a double-blind experimental study

We aimed to evaluate nasal mucosal changes and efficiency of nasal steroids and diclofenac on nasal mucosa during hyperbaric oxygen (HBO) treatment. Forty adult Albino–Wistar rats were randomized into four groups. Group 1 (control group) (n = 10) not exposed to hyperbaric or enhanced oxygen concentrations; group 2 (HBO group) (n = 10) underwent only HBO treatment; group 3 (n = 10) received HBO and intranasal mometasone furoate (10 μl/day); group 4 (n = 10) treated with HBO and diclofenac sodium (10 mg/kg/day ip). Specimens of nasal mucosa were collected after sacrificing and dissection of animals. The specimens were processed for light microscopic evaluation, and then evaluated histopathologically for fibroblastic proliferation and inflammation. Regarding the scores of inflammation, the level of inflammation in the control group was significantly less severe than the other groups (p < 0.05). Evaluation of the fibrosis scores showed that the scores of both groups 2 and 4 were significantly increased (p < 0.05). There were no statistically significant differences between groups 2, 3, and 4 as for fibrosis and inflammation (p > 0.05). Chronic HBO treatment induced mild inflammation of the nasal mucosa. These effects cannot be prevented adequately by administration of nasal steroids and diclofenac.     

肥大细胞在鼻息肉中分布特点分析

目的：探讨肥大细胞（MC）在鼻息肉中的分布特点及向上皮层移动的机制。方法：以免疫组织化学染色方法观察鼻息肉组织中MC的分布情况,观察CCL5、CCL11、CX3CL1、IL-8、IL-6等MC趋化因子在鼻息肉中的表达情况。结果：MC在鼻息肉组织中向上皮层移动,CCL5、CCL11、CX3CL1、IL-8在鼻息肉组织上皮层中的表达均明显增强;但在鼻息肉上皮层未见IL-6免疫阳性细胞,在间质的炎性细胞中偶见有免疫阳性细胞。结论：MC在鼻息肉中向上皮层移动,其移动的机制可能是上皮细胞受炎症因素刺激后CCL5、CCL11、CX3CL1、IL-8、IL-6等MC趋化因子的表达增加的原因。MC在鼻息肉的形成过程中可能发挥重要作用。     

Distribution of rantes and interleukin-5 in allergic nasal mucosa and nasal polyps.

Eosinophil-chemoattracting cytokines are thought to be important in the pathogenesis of allergic inflammation. However, little is known about the presence and significance of RANTES in nasal allergy and nasal polyps, two well-known rhinologic disorders characterized by eosinophil infiltration in the tissue. In order to evaluate the role of RANTES in eosinophil infiltration in vivo, the tissue distributions of RANTES and interleukin-5 (IL-5) and their correlation with eosinophil infiltration were investigated. Nasal mucosa specimens were obtained from 9 allergic and 12 control subjects, and nasal polyps from 6 allergic and 9 nonallergic subjects. All the subjects were divided into 4 groups: normal mucosa, allergic mucosa, nonallergic polyps, and allergic polyps. To identify the cellular localizations of RANTES and IL-5, we used specific immunohistochemical staining. We also investigated the differences in cytokine expression among the 4 groups, and the correlation between cytokine expression and eosinophil infiltration in the tissue. RANTES was expressed in the epithelium, endothelium, and some submucosal cells, while IL-5 was confined to the cells in the submucosa. Expression of both RANTES and IL-5 significantly increased in allergic mucosa and nasal polyps compared to normal mucosa; however, there was no significant difference in their expression between allergic and nonallergic polyps. Both cytokines had a significant correlation between their expression and either total or activated eosinophil numbers. The results of this study suggest that RANTES, as well as IL-5, plays a role in eosinophil recruitment in allergic nasal mucosa and nasal polyps in vivo.     

Lipopolysaccharide decreases connexin 43 expression on nasal epithelial cells in vitro

Conclusions

Decreased connexin 43 (Cx43) expression as a result of application of lipopolysaccharide (LPS) may limit the diffusion of intercellular signaling molecules that is essential to the coordinated function of neighboring cells. Therefore, it may be related to a ciliary beating defect in nasal epithelial cells and result in accumulation of harmful substances.

Objective

Gap junction intercellular communication (GJIC) is altered during inflammation in tracheal epithelial cells. Thus, we aimed to investigate whether LPS affects the expression of Cx43, the elementary protein composing the gap junction of nasal epithelial cells, in vitro.

Material and methods

LPS (Pseudomonas aeruginosa serotype 10) was applied to epithelial cells obtained from nasal polyp for 24 h in vitro. As an inflammatory indicator, IL-8 secretion was measured using a commercially available ELISA kit. Cx43 protein was detected semi-quantitatively using Western blotting.

Results

The nasal epithelial cells constitutively secreted IL-8 at a concentration of 0.45±0.03 ng/µg protein. In the presence of 10^?2 mg/ml LPS, the concentration of IL-8 was significantly increased to 0.55±0.05 ng/µg protein (n=8). Expression of the Cx43:β-actin ratio decreased in a time- and dose-dependent fashion (10^?3–10^?1 mg/ml LPS).     

The concentration and expression of IL-5 in human nasal polyp tissue]

OBJECTIVE: To study the concentration and expression of IL-5 in nasal polyp tissues and explore its significance in the micro-environment differentiation of eosinophils accumulation and clarify the conception of nasal polyposis. METHODS: The concentration and expression of IL-5 in nasal polyp tissues of 40 patients were determined by ELISA and immunohistochemistry, and inferior turbinate mucosa from patients with nasal polyps and healthy volunteers was used as control. RESULTS: 1. IL-5 concentration in the polyp tissues was significantly higher than that in inferion turbinate mucosa(P < 0.05). There was no significant difference in inferion turbinate mucosa between the patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 concentration in polyp tissues was markedly higher in patients with extensive polypoid change of nasal mucosa, history of previous polypectomy and allergic rhinitis compared with those without these features (P < 0.05). IL-5 concentration had no correlation with age and sex (P > 0.05). 2. 80.1% of the eosinophils were positive for IL-5 and 90.9% of IL-5 positive cells were eosinophils. Only 3.7% of the lymphocytes and neutrophils were IL-5 positive, and IL-5 was not detectable in epithelial cells. IL-5 expression in eosinophils of polyp tissues was remarkably stronger than that of the turbinate mucosa (P < 0.05). There was no significant difference in inferion turbinate mucosa between the patients with nasal polyps and healthy volunteers (P > 0.05). IL-5 expression of eosinophils in polyp tissues was significantly stronger in patients with extensive polypoid change of nasal mucosa, history of previous polypectomy and allergic rhinitis compared with those without these features (P < 0.05). There was no significant difference in IL-5 expression in lymphocytes and neutrophils between polyp tissues and inferior turbinate nasal mucosa (both P > 0.05). CONCLUSION: IL-5 is a key protein in eosinophilic pathologic mechanisms in nasal polyp tissues.     

Effects of intranasal steroid treatment on the presence of biofilms in non-allergic patients with chronic rhinosinusitis with nasal polyposis

Microbial biofilms have been implicated in the pathogenesis of chronic rhinosinusitis with nasal polyposis (CRSwNP). Intranasal application of corticosteroids (INCS) is a reliable option in the management of CRSwNP. INCS medication has been suspected to influence the presence and thickness of microbial biofilms and inflammatory cell patterns in CRSwNP. Two series of identical nasal polyps obtained from non-allergic patients with CRSwNP (n = 56), who underwent endoscopic sinus surgery (ESS), were processed to hematoxylin–eosin (H.E.) and Gram staining, respectively. Patients were recruited into three groups. Group A (n = 21) consisted of patients with continuous preoperative INCS treatment. In group B (n = 17), patients were never treated by INCS, while in group C (n = 18) INCS medication was stopped at least 6 months before ESS. Biofilm positivity varied from 76.4 to 88.8 % in different subject groups. These values and average thickness of biofilms did not reach statistically significant levels (Mann–Whitney’s U probe, p > 0.05) in different patient groups. In contrast, microscopic pattern and numbers of predominant inflammatory cell populations displayed obvious differences according to INCS treatment (Mann–Whitney’s U probe, p < 0.001). According to these observations, INCS treatment does not affect the presence and thickness of microbial biofilms in CRSwNP. In contrast, it has significant effects on the pattern of inflammatory cells infiltrating the subepithelial layer, which might result in beneficially altered extracellular matrix production and cytokine release.     

Trace elements in nasal polyps

The aim of our study is to evaluate the status of selenium and zinc in nasal polyp tissues and to investigate the possible role of trace elements and antioxidants including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in nasal polyps. In this study, the antioxidant enzyme and trace element levels measured in polyp tissues of 37 patients were compared with the levels measured in conchal mucosa of 27 control cases. The antioxidant enzyme and trace element levels in tissues were measured with graphite and flame spectrophotometry methods using Shimatsu UV.1601 spectrophotometer and Perkin Elmer atomic spectrometer. The mean tissue zinc and selenium levels were, respectively, 2.55 μg/g and 30.03 pg/g in patient group, 4.37 μg/g and 44.95 pg/g in control group. The mean tissue SOD and GSH-Px levels were, respectively, 4.27 and 0.69 U/mg protein in patient group, 7.09 and 0.77 U/mg protein in control group. When the measured levels in patients and control cases were compared, there were statistically significant differences between zinc, selenium, and SOD levels (P = 0.001). There was no significant difference between GSH-Px levels (P = 0.465). In conclusion, it has been revealed that the levels of zinc, selenium, and SOD in nasal polyps were significantly lower, and it may be concluded that this may have a role in the development of nasal polyps.     

Lipopolysaccharides induces MUC5AC overproduction in human nasal epithelium

Hyperproduction of mucin in the nasal epithelium is an important feature of nasal inflammatory diseases. We investigated the mechanism of lipopolysaccharides (LPS) involvement in mucin 5 subtype AC (MUC5AC) expression in human nasal epithelial cells. The primary human nasal epithelial cells were cultured in vitro, which were treated with LPS (10 nM/ml or 1 μM/ml) for 12 and 24 h. LPS-induced MUC5AC protein was determined in nasal epithelial cells. The levels of nuclear factor kappa B p65 (NF-κBp65) and its inhibitor kappa Bα (IκBα) protein were also detected, and interleukin-1β (IL-1β) mRNA was detected by real-time PCR. LPS up-regulated MUC5AC protein in human nasal epithelial cells, and we determined that the up-regulation of MUC5AC expression was due to a time- and dose-dependent degradation of IκBα protein, which resulted in the increase of NF-κBp65 nuclear translocation. Subsequently, we also determined that LPS can induce IL-1β mRNA in a time- and dose-dependent manner. These data show that LPS treatment activated NF-κB by promoting the degradation of IκBα and the nuclear localization of NF-κBp65, which induced MUC5AC overproduction.     

低氧刺激鼻息肉黏膜上皮细胞炎性因子变化的初探

目的:研究低氧刺激慢性鼻窦炎伴鼻息肉（CRSwNP）与正常鼻黏膜上皮细胞炎性因子的变化与异同,探讨其在CRSwNP发病机制中的作用。方法:选择2015年6月至2018年1月在吉林大学中日联谊医院就诊的68例CRSwNP患者,其中男性36例,女性32例,年龄（45.2±12.5）岁（ xˉ±s,下同）,患者的...     

鼻息肉中一氧化氮合酶表达及其意义

目的 :了解鼻息肉 (NP)中一氧化氮合酶 (NOS)的分布特点和活性及其在NP发病中的作用。方法 :用免疫组织化学法检测 30例NP(NP组 )及 2 0例正常鼻黏膜 (正常鼻黏膜组 )中NOS的表达 ,并用分光光度计法检测其活性。结果 :NP组NOS活性为 (4.0 79± 0 .6 5 5 )U/mg蛋白 ,正常鼻黏膜组为 (1.5 2 6± 0 .310 )U/mg蛋白 ,二者间差异有统计学意义 (P <0 .0 1) ;NP组iNOS有大量细胞表达 ,分布在黏膜上皮、腺体和血管内皮细胞以及炎症细胞中 ,与正常鼻黏膜组比较 ,差异有统计学意义 (P <0 .0 1) ;而eNOS也有表达 ,但与正常鼻黏膜组比较 ,差异无统计学意义 ;NP组i NOS表达明显高于eNOS表达 ,其差异有统计学意义 (P <0 .0 1)。结论 :NP主要表达iNOS ,分布在黏膜上皮、腺体和血管内皮细胞以及炎症细胞中 ,其产生的大量一氧化氮在NP的发病过程中可能起着重要的作用     

The intranasal Schirmer test: a preliminary study to quantify nasal secretion

Adequate secretion of the nasal mucosa is essential for normal nasal physiology. A “dry” nose is a frequent complaint of ENT patients. Measurement of secretion is currently impossible because of the absence of a recognized test. The aim of the present study was to investigate the feasibility of an intranasal Schirmer test in a large number of patients and to define standard values for nasal secretion. The test population comprised 159 healthy, non-smoking volunteers and 30 healthy smoking volunteers. All subjects were examined by nasal endoscopy for anatomic or mucosal disease. A Schirmer test strip was placed on both sides of the mucosa of the anterior nasal septum by anterior rhinoscopy. After 10 min in standardized conditions, the strip was removed and the wetted distance was measured. Active anterior rhinomanometry (ARR) and acoustic rhinometry (AR) were later performed. In the non-smoking group (n = 159), the median wetting distance of the test strip was 10.3 mm (range 3.6–35.0 mm). Age, gender, nasal geometry, and flow (according to ARR and AR) had no significant influence on nasal secretion. The test for normal distribution was negative. In the smoking group (n = 30), the median wetting distance was 8.4 mm (range 2.5–28.0 mm), significantly shorter than the wetting distance in the non-smoking group (p < 0.05). The Schirmer test offers a practical method to quantify mucosal humidification. The test is inexpensive and well tolerated by patients. In healthy people, wetting distances from 6 to 18 mm are considered normal.     

鼻息肉调节激活正常T细胞表达和分泌因子的测定及其意义

目的 探讨在鼻息肉形成过程中,上皮应答时产生调节激活正常Ｔ细胞表达和分泌因子（ｒｅｇｕｌａｔｅｄ ｕｐｏｎ ａｃｔｉｖａｔｉｏｎ,ｎｏｒｍａｌＴｃｅｌｌ ｅｘｐｒｅｓｓｅｄ ａｎｄ ｓｅｃｒｅｔｅｄ,ＲＡＮＴＥＳ）对嗜酸粒细胞趋化、移行、局部聚集的影响。方法 采用无血清原代细胞培养法培养鼾症患者下鼻甲上皮细胞和鼻息肉上皮细胞,经炎性介质ＩＬ－１β（２５ｕｇ／Ｌ,５０ｕｇ／Ｌ）刺激后收集２４ｈ和４８     

Matrix metalloproteinases and their impact on sinusal extension in chronic rhinosinusitis with nasal polyps

MMPs (metalloproteinases) and their inhibitor TIMP (tissue inhibitor of metalloproteinases) are important in tissue remodeling and have been implicated in nasal polyp pathophysiology. The objective of the present study was to evaluate gene expression of MMP-1, MMP-2, MMP-9 and TIMP-1 gene expression in nasal polyps and compare them with normal nasal mucosa. Since MMPs could induce a more prominent tissue edema, we also assessed if these gene expressions could be related to a more extensive disease. For the experimental study, 30 nasal polyp samples from patients with CRSwNP (Chronic Rhinosinusitis with Nasal Polyposis) and 19 middle turbinates (MT) from controls were obtained. Gene expression of MMP-1, MMP-2, MMP-9 and TIMP-1 was assessed by qRT-PCR. The expressions of these genes were compared between nasal polyps and controls and correlated to each individual Lund–Mackay score, MMP-1, MMP-2 and MMP-9 were significantly more expressed in nasal polyps than in controls (P < 0.005, P < 0.0001 and P < 0.05 respectively). No expression of TIMP-1 mRNA was detected. There was no correlation between disease extension evaluated by the Lund–Mackay score and MMPs gene expression. The present results demonstrate that there is a significantly higher mRNA gene expression of MMP-1, MMP-2 and MMP-9 in patients with CRSwNP than controls. However, no correlation was observed between MMPs gene expression and sinonasal CT scan extension. Level of Evidence: 2c