肺表面活性蛋白D与IL-16在变应性鼻炎及鼻息肉中的表达和意义

目的:检测肺表面活性蛋白D(SP-D)及IL-16在变应性鼻炎鼻黏膜组织和鼻息肉组织中的表达情况,探讨SP-D及IL-16在变应性鼻炎和鼻息肉发病中的作用及意义。方法:采用免疫组织化学方法分别检测15例鼻中隔偏曲伴变应性鼻炎患者下鼻甲黏膜组织(变应性鼻炎组)、15例鼻息肉组织(鼻息肉组)和15例下鼻甲黏膜组织(对照组)中SP-D及IL-16表达。结果:SP-D、IL-16在3组中均有表达,但在变应性鼻炎组和鼻息肉组的表达均明显高于对照组(均P<0.01);SP-D和IL-16在变应性鼻炎组和鼻息肉组中的表达相比较差异无统计学意义(P>0.05)。结论:SP-D及IL-16在正常组织和病变组织中均有表达。SP-D可能参与了变应性鼻炎和鼻息肉的炎症反应过程。IL-16是变应性鼻炎和鼻息肉病变中重要的嗜酸粒细胞趋化因子,并增强了嗜酸粒细胞的局部作用,参与了变应性鼻炎和鼻息肉的病变过程。     

NLRP3炎性小体在嗜酸粒细胞性鼻息肉发病及复发中的作用

目的 探讨NLRP3炎性小体在嗜酸粒细胞性鼻息肉(ENP)发病及复发中的可能作用及机制。 方法 根据HE染色,选取80例ENP(ENP组)、24例嗜酸粒细胞性复发鼻息肉(RENP)患者(RENP组)的鼻息肉组织及同期30例鼻中隔偏曲患者(对照组)的中鼻甲黏膜。所有患者嗜酸粒细胞(EOS)浸润数通过HE染色观察并计算,采用免疫组化法检测3组NLR含热蛋白结构域3(NLRP3)炎性小体、白细胞介素-1beta(IL-1β)、IL-18的表达情况,并分析ENP、RENP组中NLRP3炎性小体与EOS浸润数的相关性。 结果 IL-1β和IL-18在ENP及RENP组中的表达均高于对照组(P<0.05),但在ENP与RENP组中的表达无显著差异(P>0.05)。NLRP3炎性小体在RENP组中的表达明显高于ENP组和对照组(P<0.01),且EOS浸润数明显多于ENP组(P<0.05)和对照组(P<0.01)。NLRP3炎性小体在ENP及RENP组中的表达均与EOS浸润数呈正相关。 结论 NLRP3炎性小体、IL-1β和IL-18均与ENP发病密切相关,NLRP3炎性小体可能参与ENP的复发。     

白细胞介素13对人鼻息肉黏膜上皮细胞嗜酸性趋化因子3蛋白含量及mRNA影响的实验研究

目的　观察白细胞介素13（interleukin-13,IL-13）刺激鼻息肉黏膜上皮细胞后嗜酸性趋化因子3,即CCL26蛋白含量及其mRNA表达变化,探讨嗜酸性粒细胞 浸润鼻黏膜的机制。方法　以IL-13细胞因子刺激原代培养的人嗜酸性鼻息肉黏膜上皮细胞。用实时定量PCR法检测CCL26 mRNA的表达,ELISA检测CCL26蛋白表达。结果  与对照组相比,IL-13刺激组鼻息肉黏膜上皮细胞CCL26蛋白分泌增加,差异具有统计学意义（P <0.05）。其增高呈浓度依赖性。结论　IL-13可诱导鼻息肉黏膜上皮细胞分泌CCL26蛋白。     

嗜酸性粒细胞及IL5在上颌窦后鼻孔息肉与鼻息肉中表达的差异

目的 通过对比上颌窦后鼻孔息肉(ACP)、伴变应性鼻炎的鼻息肉(NPwAR)及不伴变应性鼻炎的鼻息肉(NPsAR)组织中嗜酸性粒细胞浸润及IL5的表达水平,探讨嗜酸性粒细胞及IL5在不同鼻腔炎性疾病发生中作用的差异。 方法 对10例ACP、9例NPwAR及14例NPsAR组织切片行H&E染色,对嗜酸性粒细胞的浸润进行计数;提取组织总RNA,通过实时荧光定量核酸扩增检测系统(QPCR)对组织中IL5的mRNA表达水平进行检测。 结果 嗜酸性粒细胞在ACP中的浸润显著低于NPwAR(P<0.001),嗜酸性粒细胞在ACP中浸润百分比的中位数低于NPsAR,但两者间差异无统计学意义;嗜酸性粒细胞在NPwAR中的表达水平显著高于NPsAR(P<0.05)。在ACP组织中IL5的mRNA表达水平显著低于NPsAR(P<0.05)和NPwAR(P=0.001),且NPsAR组织中IL5的mRNA表达水平显著低于NPwAR(P<0.05)。 结论 ACP与NP的发病机制有所不同。嗜酸性粒细胞及IL5参与了NP的发病过程,并与变应性鼻炎的发生密切相关,但在ACP的形成中可能未起到重要作用。     

鼻息肉中IL-5mRNA表达与嗜酸性粒细胞浸润和活化的相关性研究

目的:观察鼻息肉中IL-5 mRNA的表达水平,并研究其与嗜酸性粒细胞浸润和活化状态的相关性,以期加深对鼻息肉发病机制的认识.方法:采用逆转录-多聚酶链反应(RT-PCR)结合半定量分析方法,测定16例鼻息肉中IL-5mRNA的表达水平.同时分别采用chromotrope特染和免疫组化(抗EG2)法,标记组织中的嗜酸性粒细胞和活化嗜酸性粒细胞.最后进行相关性分析.结果:①16例鼻息肉组织中均有IL-5mRNA表达,而10例中鼻甲组织中只有1例显示IL-5mRNA表达.②变应性患者(n=5)和非变应性患者(n=11),IL-5/β-actin光密度比值无显著性差异(P＞0.05).IL-5/β-actin光密度比值与chromotrope 2R阳性细胞密度相关(y=-30.892+-107.530x,r=0.510,P＜0.05),与EG2阳性细胞密度的相关性不显著(r=0.376,P＞0.05).结论:IL-5mRNA在鼻息肉组织中稳定表达,表达水平与嗜酸性粒细胞的浸润程度密切相关,说明组织中IL-5的基因表达状况与嗜酸性粒细胞浸润程度之间存在密切联系.     

S-100蛋白和HLA-DR在鼻息肉组织中的表达

目的 研究糖皮质激素治疗前后S-100蛋白和HLA-DR在鼻息肉组织中的表达, 探讨二者介导的免疫反应在糖皮质激素治疗鼻息肉的作用机制。方法 收集2010年11月至2012年6月在滨州医学院烟台附属医院耳鼻喉科行鼻内镜手术治疗的鼻息肉标本74例, 其中激素治疗组37例, 未用激素治疗组37例;另取同期收治的单纯鼻中隔偏曲患者行鼻内镜下鼻中隔偏曲矫正术中切除的下鼻甲组织21例为正常对照组。采用免疫组织化学染色法检测S-100蛋白、HLA-DR在三组的表达水平。结果 S-100蛋白、HLA-DR在有鼻息肉组织中的表达明显高于正常对照组, 差异有统计学意义(P<0.05)。S-100蛋白、HLA-DR在未经治疗的鼻息肉组中的表达明显高于经激素治疗5 d的鼻息肉组, 差异亦有统计学意义(P<0.05)。结论 S-100蛋白、HLA-DR在鼻息肉组织中的高表达提示, 由二者介导的免疫反应在鼻息肉的发病中起重要作用。激素治疗5 d后鼻息肉组织中S-100蛋白、HLA-DR的表达降低, 表明激素的抗免疫机制发挥了作用。     

鼻息肉及变应性鼻炎中IL-17表达与嗜酸粒细胞浸润的意义

目的:观察鼻息肉与变应性鼻炎中IL-17表达与嗜酸粒细胞(Eos)浸润情况,探讨IL-17与Eos在其发病中作用及相关性。方法:采用免疫组织化学SP法检测鼻息肉21例、变应性鼻炎18例及单纯中隔偏曲12例中IL-17表达情况,苏木精-伊红染色下观察炎性细胞浸润并计数Eos进行比较分析。结果:①发现在变应性鼻炎鼻黏膜下固有层、鼻息肉组织上皮层及间质内Eos、中性粒细胞为主的炎性细胞及部分腺体内有IL-17表达;②检测3组中免疫组织化学IL-17阳性细胞数、IOD(IL-17阳性表达吸收度值)及苏木精-伊红染色Eos计数,发现鼻息肉与变应性鼻炎间差异无统计学意义(P>0.05),但均高于正常对照组,差异有统计学意义(P<0.05)。Eos数与IL-17阳性细胞数在鼻息肉中存在正相关性,相关系数r=0.606,P<0.01,在变应性鼻炎中两者亦有相关性,相关系数R=0.446,P<0.05。结论:①IL-17是一种新型的细胞因子,在鼻息肉组织与变应性鼻炎鼻黏膜组织中有表达,因此在发病过程中可能起调节免疫炎症反应作用,可做为深入研究鼻息肉与变应性鼻炎发病机制的指标;②推断在鼻息肉及变应性鼻炎发病中IL-17亦有趋化E...     

BRF2在慢性鼻-鼻窦炎伴鼻息肉发病过程中的作用研究

目的 研究BRF2在慢性鼻-鼻窦炎伴鼻息肉中的表达及意义。 方法 收集山东大学第二医院耳鼻咽喉头颈外科2016至2017年慢性鼻-鼻窦炎伴鼻息肉(CRSwNP)手术标本37例及单纯鼻中隔偏曲手术标本18例,采用免疫荧光染色检测BRF2在慢性鼻-鼻窦炎伴鼻息肉组织中的表达及定位;脂多糖(LPS)刺激体外培养人鼻黏膜上皮细胞后,采用免疫荧光检测BRF2在细胞中的表达及定位,Western bloting及实时荧光定量PCR检测BRF2和内质网应激相关蛋白GRP78在细胞中的表达。 结果 BRF2在慢性鼻-鼻窦炎伴鼻息肉组织中表达量上调并在胞浆中的分布增多;在体外,LPS刺激可诱导鼻黏膜上皮细胞中BRF2表达上调并在胞浆中的分布增多,并发现内质网应激相关蛋白GRP78表达上升。 结论 BRF2可能介导了内质网应激,从而参与了慢性鼻-鼻窦炎伴鼻息肉的发病过程。     

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。     

慢性鼻窦炎伴鼻息肉合并哮喘患者临床特征及转录组分析

目的　探讨慢性鼻窦炎伴鼻息肉（c h r o n i c rhinosinusitis with polyps,CRSwNP）合并哮喘（asthma,AS）患者的临床特征,并比较C R S w N P 合并A S 组（CRSwNP+AS组）组与CRSwNP不伴AS组（CRSwNPAS组）的转录组表达差异。方法　纳入北京协和医院于2016年2月～2018年2月收治的CRSwNP患者201例,比较CRSwNP+AS与CRSwNP-AS患者的临床特征差异,通过RNA-Seq分析二者转录组差异。结果　CRSwNP+AS占22.9%,CRSwNP-AS占77.1%。与CRSwNP-AS相比,CRSwNP+AS在嗜酸性粒细胞型鼻息肉、合并过敏性鼻炎或阿司匹林不耐受的比例、筛窦CT评分及CT总分、外周血及组织嗜酸性粒细胞计数及百分比上均显著高于 CRSwNP-AS组。RNA-Seq结果显示,与CRSwNP-AS组相比,CRSwNP+AS组上调的基因主要与2型免疫炎症反应及组织重塑有关,如CLCA1、COL6A5、IGHE、CCL7、CCL24、CCL26、CCR3、IL-5、IL13等。结论　CRSwNP+AS患者临床表现更重,鼻息肉组织嗜酸性粒细胞浸润更高,2型免疫反应相关基因表达更高。     

Notch通路在鼻息肉中的表达及其与调节性T细胞表达和嗜酸粒细胞浸润的相关性研究

目的探讨Notch通路在鼻息肉中的表达及其与调节性T细胞(Treg细胞)表达和嗜酸粒细胞(Eos)浸润的相关性。方法选择2012年11月至2018年8月期间在中山大学附属第三医院接受鼻内镜手术的慢性鼻窦炎(CRS)和鼻中隔偏曲的患者,分别作为CRS组和对照组。收集慢性鼻窦炎不伴鼻息肉(CRSsNP)患者(30例,男14例,女16例,年龄18~63岁)、慢性鼻窦炎伴鼻息肉(CRSwNP)患者(58例,男38例,女20例,年龄18~65岁)的窦口鼻道复合体黏膜组织、鼻息肉,对照组(29例,男19例,女10例,年龄20~57岁)患者的下鼻甲黏膜组织。通过苏木精-伊红(hematoxylin-eosin,HE)染色法观察组织中的Eos浸润情况,并将CRSwNP分为嗜酸粒细胞性慢性鼻窦炎伴息肉(Eos-CRSwNP)和非嗜酸粒细胞性慢性鼻窦炎伴息肉(non-Eos-CRSwNP)。通过实时荧光定量聚合酶链反应(quantitative real-time polymerase chain reaction,qRT-PCR)的方法检测4组患者Notch通路受体Notch-1、Notch-2、Notch-3、Notch-4及其配体Jagged-1、Jagged-2、Delta-1、Delta-3和Delta-4的表达,Th2型细胞因子[白细胞介素4(IL-4)、IL-5、IL-13]、嗜酸粒细胞阳离子蛋白(eosinophilic cationic protein,ECP)以及Treg细胞关键转录因子Foxp3的表达。采用流式细胞术检测CD4+CD25+Foxp3+Treg细胞在4组患者鼻黏膜组织中的表达。采用SPSS 20.0软件进行统计学分析,多组间均数比较采用单因素方差分析,相关性分析采用Spearman检验。结果与对照组、CRSsNP组和non-Eos-CRSwNP组相比,Eos-CRSwNP组的IL-4、IL-5、IL-13表达水平最高(F值分别为16.930、9.197、9.116,P值均<0.05);Foxp3的表达水平最低(F=2.780,P<0.05);Foxp3与ECP呈负相关(r=-0.326,P<0.05)。Eos-CRSwNP组中CD4+CD25+Foxp3+Treg细胞占比低于对照组、CRSsNP组、non-Eos-CRSwNP组(F=13.140,P<0.01)。Notch-1、Jagged-1在Eos-CRSwNP组中表达高于non-Eos-CRSwNP组,差异有统计学意义(F值分别为5.953、 6.380,P值均<0.05);且在鼻息肉组中Notch-1/Jagged-1与Foxp3表达呈负相关(r值分别为-0.611、-0.346,P值均<0.05),与IL-4、IL-5、IL-13和ECP的表达呈正相关(r值分别为0.781、0.459,0.621、0.601,0.605、0.490,0.464、0.668,P值均<0.05);在鼻息肉组中其余Notch通路受体/配体在各组间的表达无相关性。结论 Notch-1/Jagged-1通路异常激活可能参与调控Eos-CRSwNP中Treg细胞功能的抑制,以促进Th2型炎性反应及Eos浸润。     

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.     

慢性鼻窦炎伴鼻息肉的内型研究及其指导下的精准控制与治疗

慢性鼻窦炎伴鼻息肉(CRSwNP)是指病程超过12周、伴息肉形成的鼻腔鼻窦慢性炎症,是一组表型相似但内在机制存在显著差异的复杂疾病。基于其中参与的炎症因子及其病理机制的不同可将其分为1、2、3型炎症,其中2型炎症对应于嗜酸粒细胞浸润为主的CRSwNP。不同内型(endotype)的临床表现、人口学特征、治疗反应性、预后均存在差异。因此,在深入了解不同内型病理机制的基础上,可根据其临床特点建立多种间接预测模型,以优化诊断;还可在内型指导下实现症状的精准控制与治疗,以改善预后。本文就CRSwNP内型的研究进展及其指导下的精准控制与治疗作一综述。     

Impact of allergy on phenotypic and endotypic profiles of nasal polyposis

ObjectivesTo assess the impact of allergy on clinical presentations (phenotypes) and inflammatory patterns (endotypes) of chronic rhinosinusitis with nasal polyps (CRSwNP).MethodsA single-center prospective study was conducted over an 18-month period. Fifty-seven patients with refractory CRSwNP were included. The diagnosis of allergy was based on concordant skin prick tests and symptoms. Phenotypes were determined on symptom severity score, polyp size classification and Lund-Mackay CT staging. Inflammatory endotypes were determined on biomarker analysis (IgE, IgA, IL-5, IL-9, ECP, EDN) in blood and nasal secretions. Eosinophil counts were obtained in blood, nasal secretions and polyps.ResultsPhenotype and endotype profiles were comparable in patients with (n = 15) or without (n = 42) allergy. Only asthma with high total IgE blood concentration showed association with allergy.ConclusionsThe present results suggest that allergy is not directly involved in the clinical expression and specific inflammatory pathways of CRSwNP. New therapies target inflammation signaling pathways, and identifying accurate blood and tissue biomarkers will be the line of research most likely to improve treatment of CRSwNP.     

Effect of Obstructive Sleep Apnea on Immunity in Cases of Chronic Rhinosinusitis With Nasal Polyps

Objectives.Chronic rhinosinusitis with nasal polyps (CRSwNP) is a more severe inflammatory form of CRS that often coexists with obstructive sleep apnea (OSA). However, little is known about the relationship between OSA and the immune profile in patients with CRSwNP. We aimed to investigate the immune profile of patients with CRSwNP according to OSA severity.Methods.This study included 63 patients with CRSwNP and nine control subjects. Protein levels of inflammatory mediators were determined using multiplex immunoassays. All patients underwent standard polysomnography.Results.In patients with eosinophilic CRSwNP (ECRSwNP), interleukin (IL)-6 and chemokine [C-X-C motif] ligand (CXCL)-1 (type 1 immune-related markers) were upregulated in cases of moderate-to-severe OSA. Additionally, IL-4, IL-13, C-C motif chemokine (CCL)-11, CCL-24 (type 2 immune-related markers), and IL-17A (a type 3 immune-related marker) were present at elevated levels in patients with moderate-to-severe OSA. Although there were no significant differences in type 1, 2, or 3 immune-related markers among patients with non-eosinophilic CRSwNP (NECRSwNP) according to the severity of OSA, transforming growth factor-beta expression was higher in those with moderate-to-severe OSA. Furthermore, in ECRSwNP with moderate-to-severe OSA, associations were detected between serum markers and some upregulated inflammatory markers.Conclusion.OSA may increase the heterogeneity of the immune profile (types 1, 2, and 3) in patients with ECRSwNP, but not in those with NECRSwNP.     

Th2 cytokines associated with chronic rhinosinusitis with polyps down-regulate the antimicrobial immune function of human sinonasal epithelial cells

BACKGROUND: Chronic rhinosinusitis with nasal polyps (CRSwNPs) is a disorder characterized by persistent eosinophilic Th2 inflammation and frequent sinonasal microbial colonization. It has been postulated that an abnormal mucosal immune response underlies disease pathogenesis. The relationship between Th2 inflammatory cytokines and the innate immune function of sinonasal epithelial cells (SNECs) has not been explored. METHODS: Human SNECs (HSNECs) isolated from control subjects and patients with CRS were assessed for expression of antimicrobial innate immune genes and proinflammatory cytokine genes by real-time polymerase chain reaction, ELISA, and flow cytometry. A model of the Th2 inflammatory environment was created by exposure of primary HSNEC to the Th2 cytokine interleukin (IL)-4 or IL-13 for 36 hours, with subsequent assessment of innate immune gene expression. RESULTS: HSNEC obtained from CRSwNP patients displayed decreased expression of multiple antimicrobial innate immune markers, including toll-like receptor 9, human beta-defensin 2, and surfactant protein A. Baseline expression of these genes by normal and CRS HSNEC in culture is significantly down-regulated after incubation with IL-4 or IL-13. CONCLUSION: Expression of multiple innate immune genes by HSNEC is reduced in CRSwNP. One mechanism appears to be a direct effect of the leukocyte-derived Th2 cytokines present in the sinonasal mucosa in CRSwNP. Impaired mucosal innate immunity may contribute to microbial colonization and abnormal immune responses associated with CRSwNP.     

Increased expression of nucleophosmin is associated with the pathophysiology of chronic rhinosinusitis with nasal polyposis

ObjectiveNucleophosmin (NPM1) has been suggested to be involved in the pathophysiologic mechanism of inflammatory disorders. We measured the expression level of NPM1 in nasal polyp (NP) tissues of patients with chronic rhinosinusitis with nasal polyposis (CRSwNP). We also assessed the correlation between NPM1 expression and other parameters such as eosinophilic infiltration, inflammatory cytokines, and clinical indicators such as Lund-Mackay computed tomography (CT) score.MethodsThirty patients with CRSwNP were included. We performed pre-operative CT scan to determine Lund-Mackay CT scores. During endoscopic sinus surgery, we harvested NP tissues from patients with CRSwNP. We performed Sirius red staining to evaluate eosinophilia and conducted immunohistochemical staining for NPM1 and real-time PCR for cytokines including interleukin (IL)-5, IL-17A, and IL-32.ResultsThe mRNA expression of NPM1 was significantly up-regulated in eosinophilic NP tissues (RQ 0.58 ± 0.06), compared to non-eosinophilic NP tissues (RQ 0.38 ± 0.08, p < 0.05). In the epithelium of NP tissue, a significant positive correlation was observed between eosinophilic infiltration and NPM1 expression. The expression of NPM1 was significantly correlated with that of IL-5 (r = 0.6229, p = 0.0004), IL-17A (r = 0.5971, p = 0.001), and IL-32 (r = −0.5985, p = 0.0068). There was no significant correlation between the mRNA expression of NPM1 and the Lund-Mackay CT score (Spearman r = −0.2563, p = 0.1879).ConclusionExpression of NPM1 was significantly increased in eosinophilic NP tissues from patients with CRSwNP. We observed an association between NPM1 expression and various pro-inflammatory cytokines such as IL-5, IL-17, and IL-32 and eosinophilic infiltration, which is thought to contribute to the pathophysiology of NP.     

Overexpression of hypoxia-inducible factor 1α is associated with neutrophilic inflammation in chronic rhinosinusitis with nasal polyps

ObjectiveThis study aimed to assess the possible role of hypoxia-inducible factor 1α (HIF-1α) in promoting neutrophilic inflammation in chronic rhinosinusitis with nasal polyps (CRSwNP) patients.MethodsWe examined HIF-1α expression in sinonasal tissues from CRSwNP patients and healthy controls by using immunohistochemistry, qRT-PCR, and western blot. Next, the stimulatory effects of several cytokines (IFN-γ, IL-17A, IL-6, etc.) and reagents (dexamethasone (DEX), clarithromycin (CAM) and curcumin (CUM)) on HIF-1α expression in cultured normal nasal epithelial cells (NECs) were also evaluated. Moreover, the effects of CAM and glucocorticoid on nasal symptoms and signs of uncontrolled neutrophilic CRSwNP patients were evaluated.ResultsThe mRNA and protein expression of HIF-1α were significantly increased in polyp tissues compared with healthy controls (P < 0.05), and the HIF-1α level in polyp tissues was positively associated with IL-17A production and tissue neutrophilia (P < 0.05). Moreover, in cultured NECs, HIF-1α expression was upregulated in the presence of IL-17A and IL-6 (P < 0.05). Both CAM and CUM showed an additive effect with DEX in inhibiting HIF-1α expression (P < 0.05). Moreover, combined glucocorticoid and CAM significantly improved nasal symptoms and signs compared with glucocorticoid alone in uncontrolled neutrophilic CRSwNP patients (P < 0.05).ConclusionOur findings indicate that HIF-1α is associated with neutrophilic inflammation and glucocorticoid resistance in CRSwNP patients.     

Interferon-α action in cytokine profile in eosinophilic nasal polyp cultures

IntroductionChronic rhinosinusitis is currently classified into two types: chronic rhinosinusitis without nasal polyps and chronic rhinosinusitis with nasal polyps. In the West, approximately 80% of chronic rhinosinusitis with nasal polyps cases are characterized by a predominantly eosinophilic cell infiltrate and a Th2 cytokine pattern.ObjectiveTo evaluate the effect of Interferon-α on cytokine levels of the eosinophilic nasal polyp cell culture supernatant.MethodsCell cultures were performed based on nasal polypoid tissue samples collected from 13 patients with eosinophilic chronic rhinosinusitis with nasal polyps. Polyps were considered eosinophilic according to the histopathological examination. Cell cultures were stimulated with 3000 IU of interferon-α. Before and after the stimulus, concentrations of Interferon-γ, tumor necrosis factor αand IL 2, 4, 6 and 10, using cytometric bead array, were assessed.ResultsCell samples from eosinophilic nasal polyps from 13 patients were included in the study. Twenty-four hours after interferon-α stimulation, eosinophilic nasal polyp culture supernatants showed significantly decreased IL-4 concentrations and increase in interferon-γ, IL-10 and IL-6 concentrations compared to controls. There were no significant differences in tumor necrosis factor -α and IL-2 concentrations.ConclusionWe demonstrated that interferon-α in vitro alters the pattern of cytokines in cell cultures of eosinophilic nasal polyps. Analysis of these alterations suggests that interferon-α promotes a rebalancing of inflammatory profiles in cell cultures, favoring the expression of Th1 and regulatory cytokines over Th2 cytokines.     

鼻息肉组织及上皮细胞的形态学特征与嗜酸粒细胞阳离子蛋白的表达

目的:研究鼻息肉组织及其上皮细胞的形态学特征,检测嗜酸粒细胞阳离子蛋白(ECP)在其中的表达,探讨其发病机制及病理特点、ECP的作用。方法:慢性鼻-鼻窦炎不伴鼻息肉(鼻窦炎组)5例,慢性鼻-鼻窦炎伴鼻息肉(鼻息肉组)5例,正常下鼻甲黏膜(对照组)5例,标本均分成2份,一份放入4%的戊二醛液中固定,常规电镜制片;另一份放入10%甲醛中固定,石蜡包埋行ECP原位杂交检测。结果:电镜下见鼻息肉组黏膜上皮排列紊乱,基底膜增厚,纤毛排列不整齐,倒伏,其中基底层有脱颗粒现象,鼻窦炎组上皮层破坏、基底膜水肿;对照组上皮层连续,基底层排列整齐。鼻窦炎组、鼻息肉组ECP阳性表达于上皮层、黏膜下层及血管周,呈棕褐色,但鼻窦炎组表达较鼻息肉组低。结论:鼻息肉组织及上皮细胞内有活化的嗜酸粒细胞,脱颗粒现象,鼻息肉组织中ECP高表达,说明鼻窦炎和(或)鼻息肉组织中有活跃的炎症反应,引起组织上皮损伤和脱落,最终形成鼻息肉。