鼻息肉中IgE及GM-CSF与嗜酸粒细胞浸润的相关性研究

目的:探讨鼻息肉组织中IgE和粒细胞巨噬细胞集落刺激因子(GM-CSF)的表达对嗜酸粒细胞浸润聚集的作用及二者的关系及意义。方法:采用免疫组织化学SP法检测30例鼻息肉标本(鼻息肉组)和11例下鼻甲部分切除术患者下鼻甲黏膜标本(对照组)组织中IgE阳性细胞分布情况;酶联免疫吸附法检测上述标本中GM-CSF含量,同时观测2组组织中嗜酸粒细胞的浸润程度。结果:鼻息肉组中IgE、GM-CSF水平及嗜酸粒细胞数均明显高于对照组(均P<0.01),且IgE与GM-CSF的水平分别与鼻息肉组织中嗜酸粒细胞浸润数呈正相关(r=0.65、0.62,均P<0.01),鼻息肉组中IgE与GM-CSF水平呈正相关(r=0.52,P<0.01)。结论:鼻息肉是以嗜酸粒细胞浸润为特征的疾病过程,IgE及GM-CSF则对鼻息肉中嗜酸粒细胞浸润发挥重要作用,IgE介导的Ⅰ型变态反应在鼻息肉的形成和发展中占有重要地位。     

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.     

鼻息肉中嗜酸性粒细胞浸润和活化与白细胞介素5表达

目的通过观察鼻息肉中嗜酸性粒细胞的浸润和活化状态,以及白细胞介素5(interleukin-5,IL-5)的表达水平,揭示二者的关系,探索鼻息肉中嗜酸性粒细胞浸润增多调节的机制。方法对30例鼻息肉组织,分别采用chromotrope2R法标记鼻息肉组织中的嗜酸性粒细胞,采用免疫组织化学ABC法,观察嗜酸性粒细胞的活化状态和IL-5的表达水平,然后进行相关性分析。结果①chromotrope 2R染色可特异性地将嗜酸粒细胞的胞浆染成粉红色,活化嗜酸性粒细胞(EG2阳性细胞)和IL-5阳性细胞胞浆中可见棕褐色颗粒。30例鼻息肉中嗜酸性粒细胞的密度为50.7±16.1/0.25mm2,活化嗜酸性粒细胞的密度为20.7±14.3/0.25mm2,IL-5阳性细胞密度为15.9±5.7/0.25mm2,上述三项指标在变应性患者和非变应性患者比较未见显著性差异(P>0.05);②鼻息肉中IL-5阳性细胞密度与chromotrope2R阳性细胞密度密切相关(y=14.723 2.012x,r=0.642,P<0.01),与EG2阳性细胞密度也存在相关关系(y=11.817 1.092x,r=0.602,P<0.05)。结论嗜酸性粒细胞是鼻息肉组织中IL-5的来源之一,IL-5的表达水平是鼻息肉中嗜酸性粒细胞的浸润和活化的调节因素之一。     

鼻腔应用布地奈德对鼻息肉中IL-5表达的作用

目的 :观察鼻腔应用布地奈德治疗对鼻息肉中 IL - 5表达的影响 ,深入了解糖皮质激素类药物局部治疗对鼻息肉的作用机制。方法 :采用免疫组化 ABC法 ,观察经布地奈德治疗 6～ 8周和未经治疗的鼻息肉组织 (各 16例 )中 ,IL -5阳性细胞的浸润和分布状况。结果 :鼻息肉组织固有层中可见 IL - 5阳性细胞表达 ,且多为嗜酸性粒细胞 ,IL - 5阳性细胞密度与嗜酸性粒细胞的浸润程度密切相关 (y=14.782 + 2 .0 0 2 x,r=0 .6 48,P <0 .0 1)。经布地奈德治疗的鼻息肉组织中 IL - 5阳性细胞的浸润程度有下降的趋势 ,但与未治组织的差异未达显著性水平 (P >0 .0 5 )。结论 :1IL - 5的表达与嗜酸性粒细胞的浸润密切相关 ,嗜酸性粒细胞是鼻息肉组织中 IL- 5的来源之一 ;2鼻腔应用布地奈德 6～ 8周治疗可能抑制鼻息肉中 IL- 5表达。     

鼻腔局部应用激素对鼻息肉中嗜酸粒细胞浸润和活化的影响

目的　探讨鼻腔局部应用激素布地奈德 (budisonide)对鼻息肉组织中嗜酸粒细胞浸润和活化状态的影响。方法　采用免疫组化ABC法和chromotrope 2R染色 ,分别标记鼻息肉组织中的活化嗜酸粒细胞和全部嗜酸粒细胞。观察比较经布地奈德喷鼻治疗 6～ 8周和未经治疗的鼻息肉组织 (各16例 )中 ,嗜酸粒细胞浸润和活化状况。结果　①鼻息肉组织中浸润的嗜酸粒细胞多处于活化状态 ;②激素治疗组与未治组相比 ,前者鼻息肉组织中嗜酸粒细胞的活化比例显著减少 (P <0 0 1) ,而嗜酸粒细胞总数和活化嗜酸粒细胞的浸润程度虽呈下降趋势 ,组间比较差异未见显著性 (P >0 0 5 )。结论　嗜酸粒细胞的浸润和活化程度在鼻息肉的发病机制中可能起重要作用。 6～ 8周布地奈德喷鼻治疗使局部嗜酸粒细胞性炎性反应的严重程度减轻 ,其对嗜酸粒细胞活化水平的抑制作用 ,较其对嗜酸粒细胞数量的影响更为显著。     

鼻息肉中总IgE及IL-5与嗜酸性粒细胞浸润的相关性研究

目的探讨鼻息肉中总IgE、IL-5的表达、嗜酸性粒细胞(eosinophils,EOS)浸润情况及其相关关系,以分析其在鼻息肉发病中的作用.方法采用免疫组化SP法检测38例鼻息肉,15例正常鼻腔黏膜组织中总IgE阳性细胞分布情况;酶联免疫吸附试验(ELISA)检测鼻息肉中IL-5含量;Chromotrope 2R特染法检测鼻息肉中EOS浸润情况.结果①鼻息肉组织中总IgE、IL-5及EOS水平均明显高于正常鼻腔黏膜组织,P均＜0.01;②鼻息肉组织中总IgE与EOS、IL-5与EOS以及IL-5与总IgE之间均呈明显正相关关系,相关系数r分别为0.843,0.642,0.667,P均＜0.01.结论EOS浸润在鼻息肉的形成和发展中起重要作用,而IgE及IL-5则对鼻息肉中EOS浸润发挥重要影响.     

两种细胞粘附分子及肿瘤坏死因子-α在鼻息肉组织中的表达及意义

目的 :通过检测鼻息肉组织中细胞间粘附分子 (ICAM- 1) ,血管细胞粘附分子 (VCAM- 1)及肿瘤坏死因子(TNF-α)的表达 ,以了解三者与嗜酸性粒细胞浸润的关系 ,探讨粘附分子与 TNF-α的相关性。方法 :分别以 ICAM- 1、VCAM- 1及 TNF-α的单克隆抗体对 4 2例鼻息肉组织 (鼻息肉组 )和 16例鼻甲粘膜 (对照组 )进行免疫组织化学染色 ,光镜观察。结果 :ICAM- 1、VCAM- 1及 TNF-α在鼻息肉组织中的表达均明显高于对照组 (P <0 .0 1) ,ICAM- 1、VCAM- 1与 TNF-α的表达呈正相关 (r1 =0 .5 2 ,p1 <0 .0 1;r2 =0 .6 8,p2 <0 .0 1)。鼻息肉中两种细胞粘附分子与嗜酸性粒细胞浸润程度一致。结论 :1嗜酸性粒细胞在鼻息肉的发生发展中发挥重要作用 ;2而 TNF-α通过上调鼻息肉组织内皮细胞ICAM- 1及 VCAM- 1的表达 ,从而促进嗜酸性粒细胞与内皮细胞的粘附 ,穿内皮迁移、聚集于组织局部 ,引起鼻息肉形成。     

慢性鼻窦炎伴鼻息肉中IL4基因异常甲基化与嗜酸性粒细胞浸润机制研究CSCD

目的 探究IL4基因异常甲基化与嗜酸性粒细胞浸润的机制。方法 选取符合慢性鼻窦炎伴鼻息肉诊断标准的9例患者标本,根据鼻息肉组织病理嗜酸性粒细胞(Eos)浸润程度,分为嗜酸粒细胞性慢性鼻窦炎伴鼻息肉(eosinophilic chronic rhinosinusitis with nasal polyps,ECRSwNP)和非嗜酸粒细胞性慢性鼻窦炎伴鼻息肉(non-ECRSwNP,NECRSwNP)两组,比较两组样本不同炎症细胞计数差异。设计DNA甲基化引物,MassARRAY法进行IL4基因CpG位点DNA甲基化检测。运用线性拟合回归探究Eos计数和IL4基因CpG位点甲基化程度之间的线性关系。结果 ECRSwNP组Eos计数高于NECRSwNP组(P=0.0043),其他炎症细胞数目相比无统计学差异。DNA甲基化检测显示,ECRSwNP组CpG位点5(CpG-5)的甲基化率显著低于NECRSwNP组(P=0.0311)。我们还发现IL4基因CpG-5甲基化率和组织Eos浸润有着较强的负相关关系(R2=0.53,P=0.0261)。通过回归模型拟合得到Eos=496.25-5.13×CpG-5(DNA甲基化率),结果表明随着IL4基因CpG-5甲基化水平的降低,鼻息肉中Eos浸润增加。结论 IL4基因异常甲基化状态与Eos浸润程度相关,为CRSwNP发病机制提供表观遗传学证据。     

IL-17在鼻息肉中的表达及其意义

目的：研究伴变应性体质和不伴变应性体质的鼻息肉患者息肉中IL-17的表达及与病理特征的关系。方法：30例鼻息肉患者根据皮肤点刺实验分为伴变应性体质组和不伴变应性体质组,通过苏木精一伊红染色法嗜酸粒细胞浸润程度评价组织学特征,运用ELISA及RT—PCR方法检测IL-17在鼻息肉中的表达。结果：伴变应性体质组嗜酸粒细胞浸润明显高于不伴变应性体质组（P〈O．01）;两组息肉组织中IL-17蛋白及mRNA水平均显著高于对照组（P〈O．01或P〈0．05）,且变应性体质组IL-17水平明显高于不伴变应性体质组,鼻息肉患者IL-17水平与嗜酸粒细胞浸润呈正相关。结论：IL-17在鼻息肉组织中表达明显上调,且在伴变应性体质组中水平更高,提示IL-17可能在鼻息肉的发病机制中发挥重要作用,变应性因素在鼻息肉形成中可能是通过促进IL-17曲立牛而,卉＇枷曲     

鼻息肉中总IgE与嗜酸粒细胞浸润的相关性研究

目的:探讨总IgE及嗜酸粒细胞(EOS)在鼻息肉中的表达及其相关关系,以分析其在鼻息肉发病中的作用。方法:采用免疫组织化学SP法检测38例鼻息肉、15例中鼻甲黏膜组织中总IgE阳性细胞表达;采用Chromotrope2R染色法检测EOS浸润情况。结果:①鼻息肉组织中总IgE及EOS水平均明显高于中鼻甲黏膜组织,均P<0.01;②鼻息肉组织中总IgE与EOS呈明显正相关关系(r=0.843,P<0.01)。结论:EOS浸润在鼻息肉的形成和发展中起重要作用,IgE介导的Ⅰ型变态反应对鼻息肉中EOS浸润发挥一定作用。     

鼻息肉组织中白细胞介素—5和粒细胞巨噬细胞集落刺激因子水平测定及其意义

OBJECTIVE: To study the relationship between the concentration of interleukin-5 (IL-5) and granulocyte-macrophage colony-stimulation factor (GM-CSF) in nasal polyps and its significance. METHODS: The concentration of IL-5, GM-CSF was measured by enzyme linked immunosorbent assay (ELISA) in a total of 38 cases, and the number of eosinophils were counted in the same tissues. RESULTS: The statistical analysis showed that the concentrations of IL-5 and GM-CSF have significant difference (P < 0.05) between the nasal polyps and the control group. There is a positive correlation between the quality of IL-5/GM-CSF and the extent of eosinophils (r = 0.75/0.71). CONCLUSION: This study demonstrated that there are inductive compartments, namely the microenvironment, in which immune-effector and inflammatory cell function is modulated in nasal polyps. A special discussion has been made of eosinophils and IL-5/GM-CSF in the formation of nasal polyps, and the effect of the IL-5/GM-CSF in the accumulation of eosinophils.     

鼻息肉组织中IL-4 IL-5 IL-6和IL-8的含量及其表达

目的：通过检测鼻息肉组织中IL-4、IL-5、IL-6、IL-8的水平及表达方式,探讨细胞因子在鼻息肉形成中的作用。方法：应用放射免疫法检测54例鼻息肉组织（鼻息肉组）中IL-4、IL-5、IL-6及IL-8的浓度,同时用免疫组织化学法观察其表达,以22例行鼻中隔手术患者中鼻甲黏膜作为对照（对照组）。结果：鼻息肉组IL-5及IL-8水平均明显高于对照组（均P〈0．01）;而IL-6在两组间差异无统计学意义（P〉0．05）;IL-4在变应原皮试阳性组明显增加,与对照组比较P〈0．05。IL-4主要表达于息肉组织内炎性细胞,多为淋巴细胞或浆细胞;IL-5主要表达于鼻息肉组织中嗜酸粒细胞和淋巴细胞;IL-6、IL-8主要表达于息肉上皮层及息肉组织中的炎性细胞,皮试阳性组和阴性组间无明显区别。结论：IL-5及IL-8在所有鼻息肉组织中具有一定作用,而IL-4仅在变应原皮试阳性息肉中具有一定意义,IL-6在鼻息肉组织中无明显作用。     

白介素-5在变应性鼻炎鼻黏膜中的表达及意义

目的：了解白介素 5（IL 5）在变应性鼻炎鼻粘膜内浸润的炎性细胞中的表达及分布，探讨IL 5与嗜酸性粒细胞积聚及发病机制的关系。方法：采用免疫组化染色法（SP法）对变应性鼻炎下鼻甲粘膜（A组）、单发鼻息肉组织（B组）、无变应性鼻炎下鼻甲粘膜组织（C组）切片进行IL 5染色，对IL 5染色阳性细胞计数分类，对统计结果行方差分析。结果：A、B两组组织中可见较多的IL 5阳性染色细胞，多见嗜酸性细胞、淋巴细胞、中性粒细胞等阳性染色，两组IL 5阳性细胞总数、Eos计数无统计学差异，但均高于C组（P＜0.01）。结论：IL 5在变应性鼻炎鼻粘膜、鼻息肉组织内浸润的多种炎性细胞中表达，能客观反映免疫或炎症反应的程度，可作为变应性鼻炎诊断评分系统的补充。     

白细胞介素-5与嗜酸性粒细胞阳离子蛋白在鼻息肉发病中的意义及相互关系

目的：探讨白细胞介素—5(IL—5)及嗜酸性粒细胞阳离子蛋白(ECP)在鼻息肉发病中的作用及相互关系。方法：采用pharmacia CAP荧光免疫系统和ELISA双抗体夹心法对30例鼻息肉患者(鼻息肉组)和8例鼻中隔偏曲或阻塞性睡眠呼吸暂停综合征(OSAS)患者(对照组)分别进行血清中ECP及组织匀浆中ECP、IL—5的检测。结果：鼻息肉组匀浆中IL—5的水平明显高于对照组，其差异有显著性意义(P＜0．05)；鼻息肉组血清与匀浆中的ECP含量明显高于对照组，其差异亦有显著性意义(P＜0．05)。鼻息肉组匀浆中IL—5与血清中ECP水平呈明显正相关(r＝0．598，P＜0．05)；与匀浆中的ECP水平也呈正相关(r＝0．451，P＜0．05)。结论：ECP是嗜酸性粒细胞活化的标志，也是导致鼻腔炎症发生的重要因子；IL—5在鼻息肉组织中高表达，并与血清和组织中ECP水平密切相关，共同促进鼻腔炎症过程的不断加重。     

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.     

Eosinophils in nasal polyposis pathogenia. Bibliographic revision

The aim of this work is to do a review on relations between the eosinophils and nasosinusal polyps. The etiopathogenical action of eosinophils are reviewed, from bone narrow to nasal mucosa infiltration. The mechanism of tissue eosinophilic mediated inflammation, his regulation and the significance of the balance between positive (IL-5) and negative (TGFbeta and IL10) mediators are discussed. We conclude that the degree of eosinophilic infiltration and degranulation's patterns are important aspects conditioning the severity and prognosis of chronic rhinosinusitis and nasosinusal polyps.     

Inflammatory cells in nasal mucosa and nasal polyps

OBJECTIVE: Since some controversy exists concerning the frequency of inflammatory cells in nasal polyps, we have compared the frequency of tissue inflammatory cells (lymphocytes, neutrophils, eosinophils and plasma cells) including 11 kinds of lymphocyte subsets in the same specimens of nasal mucosa and nasal polyps. METHODS: Histopathological observations and flow cytometric analyses were performed on eight mucosal specimens of the inferior turbinates of patients with nasal polyps and on 13 polyp specimens. RESULTS: Nasal polyps contained significantly more eosinophils, neutrophils and plasma cells than nasal mucosa, and EG2+ cells (activated eosinophils) were significantly more frequent in nasal polyps than in nasal mucosa. Flow cytometric analysis showed that there were no significant differences in the frequencies of lymphocytes and lymphocyte subsets (CD1+, CD2+, CD3+, CD5+, CD7+, CD4+, CD8+, CD10+, CD19+, CD20+ and HLA-DR+ cells) including CD4/8 ratios between nasal mucosa and polyps, though, both nasal mucosa and polyps contained significantly more lymphocytes than eosinophils, neutrophils or plasma cells. The T cell lineage (CD2+, CD3+, CD5+ and CD7+ cells) was found in high frequency and B cell lineage (CD10+, CD19+ and CD20+ cells) in low frequency in both nasal mucosa and polyps. The frequency of HLA-DR+ cells (most of which were activated T cells) was not significantly different between nasal mucosa and nasal polyps. CONCLUSION: Histopathological and flow cytometric analyses were performed on the composition of inflammatory cells in nasal mucosa of the inferior turbinates and in polyps from the same patients. The elevated numbers of activated eosinophils, neutrophils and plasma cells in nasal polyps compared with nasal mucosa suggest that inflammatory processes play important roles in the pathophysiology of nasal polyps. The frequencies of lymphocytes and lymphocyte subsets were not significantly different between these two tissues.     

鼻息肉中碳酸酐酶IX表达与嗜酸细胞浸润相关性分析

目的研究碳酸酐酶IX（carbonic anhydrase IX,CAIX）在鼻息肉组织中表达及其与嗜酸细胞（eosinophils,Eos）浸润的相关性,探讨CAIX在鼻息肉发病中的作用。方法收集28例鼻息肉组织（鼻息肉组）和12例正常下鼻甲组织（对照组）,HE染色观察Eos浸润情况,免疫组化检测CAIX的表达。结果与对照组比较,鼻息肉组Eos浸润显著增加,CAIX免疫阳性细胞数及着色强度明显增高。图像分析显示,鼻息肉组CAIX的积分光密度（103/HP）为26.87±4.60,与对照组3.25±1.20相比,有显著性差异（P〈0.05）;Spearman等级相关分析显示,鼻息肉中CAIX表达与Eos浸润表达密切相关（r=0.632,P〈0.01）。结论鼻息肉组织中CAIX表达、Eos浸润均上调,二者有协同表达关系,说明鼻息肉组织中存在缺氧及缺氧适应现象,CAIX表达上调,从而使细胞适应缺氧微环境,保证缺氧区域细胞的持续生长和增殖,促进Eos浸润生长,导致一种慢性炎症反应。     

Effect of erythromycin on the expression of apoptotic gene on eosinophils in nasal polyps in vitro]

OBJECTIVE: To study the expression of apoptotic gene in nasal polyps and the possible effect of erythromycin on it. METHODS: Total 40 nasal polyps biopsies were detected the expression of Bcl-2, Bax, Bcl-xL as well as Fas protein on eosinophils by immunohistochemistry. The expression of Bax, Bcl-xL, Fas on eosinophils in 15 nasal polyps explants which cultured with or without erythromycin were detected also. RESULTS: In nasal polyps, the expression of Bcl-2, Bax, Bcl-xL and Fas protein on eosinophils are 5% (2/10), 65% (26/40), 40% (16/40) and 90% (36/40) respectively. As compared with inferior turbinate, there were significant difference in expression of Bax and Fas protein between two tissues (P < 0.05). After cultured with erythromycin, the expression of Bax protein increased (P < 0.05), but the expression of Bcl-xL and Fas protein had not changed. CONCLUSION: Bax may be the most significant gene which control the apoptosis of eosinophils in nasal polyps. And a possible mechaism which erythromycin promote the apoptosis of eosinophils is increase the expression of Bax.