Clinical factors influencing the eosinophil infiltration of nasal polyps

AIMS AND METHODS: Our study, based on a retrospective chart analysis, was aimed 1) to describe the varying degree of eosinophil infiltration in a series of 263 adult patients operated on diffuse and bilateral nasal polyposis (NPS) after failure of medical treatment, in 15 cystic fibrosis patients with bilateral nasal polyps, and in 31 patients with chronic sinusitis without polyps (18 bilateral, 13 unilateral) 2) to search for clinical factors that might influence the degree of eosinophil infiltration. Eosinophil infiltration was expressed semi-quantativity as a percentage of inflammatory cells. RESULTS: Our study confirms that eosinophil infiltration is a striking feature of nasal polyposis. All patients with chronic sinusitis showed less than 10% eosinophils (mean +/- SEM = 2 +/- 2%) whereas 88% of patients with NPS showed more than 10% eosinophils (50 +/- 2%). Cystic fibrosis lied in between with 40% of patients showing more than 10% eosinophils. In idiopathic bilateral NPS the number of eosinophils was increased in patients with asthma (58 +/- 3%) and even more in Widal's triad (75 +/- 4%). Atopic patients did not have more eosinophils (52 +/- 5%). Patients treated with systemic steroids within two months before surgery showed decreased eosinophil infiltration (22 +/- 3% vs 50 +/- 2 for treated versus untreated) whereas patients treated with topical steroids did not (47 +/- 2%). CONCLUSIONS: Thus, a link might exist between clinical presentation and eosinophil infiltration. Chronic sinusitis and nasal polyps are probably not the same disease. Eosinophils appear as a link between nasal polyps, asthma and aspirin intolerance. Atopic status does not modify eosinophil infiltration of nasal polyps. Systemic steroids appear significantly more effective to reduce the eosinophil infiltrate than topical steroids in our selected group of operated patients.     

TNF-alpha upregulates VCAM-1 and NF-kappaB in fibroblasts from nasal polyps

OBJECTIVE: Lung and synovial fibroblasts produce VCAM-1 in response to TNF-alpha. However, the massive infiltration of eosinophils, the effects of the increased amount of TNF-alpha and the production of VCAM-1 in human nasal polyp fibroblasts are not yet fully understood. The present study examines the role of VCAM-1 and the molecular mechanism of its expression in nasal fibroblasts. METHODS: Nasal fibroblasts were isolated from human nasal polyps and after four passages, the cells were stimulated with TNF-alpha and VCAM-1 expression was examined by ELISA, flow cytometry, and RT-PCR. The activation of NF-kappaB induced by TNF-alpha was determined by electrophoretic mobility shift assays and the influence on the expression of VCAM-1 was investigated. RESULTS: VCAM-1 protein and mRNA were expressed in unstimulated controls and remarkably increased by TNF-alpha stimulation. NF-kappaB activity was enhanced by TNF-alpha stimulation and remarkably suppressed by NF-kappaB proteasome inhibitor. CONCLUSIONS: The present study discovered that nasal fibroblasts produce VCAM-1 protein and mRNA and that production is increased by TNF-alpha stimulation. Furthermore, VCAM-1 expression in nasal fibroblasts is induced through an NF-kappaB-dependent pathway. These findings might provide a rationale for using NF-kappaB inhibitors as a treatment for nasal inflammatory diseases such as polyps.     

Decreased connexin 43 expression correlated with eosinophil infiltration in nasal polyps

BACKGROUND: A nasal polyp usually is characterized by eosinophil infiltration. Eosinophil-fibroblast interaction is an important event of persistent inflammation in airways. We have found abundant connexin 43 (Cx43) expression in subepithelial fibroblasts of nasal mucosa. Thus, we aim to analyze the relationship of Cx43 expression and eosinophil in nasal polyps. METHODS: In 25 nasal polyps and 19 inferior turbinates, indirect immunofluorescent and hematoxylin and eosin staining were performed in adjacent sections. We calculated the density of Cx43 staining and eosinophil individually by fluorescent and light microscope. RESULTS: Positive Cx43 staining under confocal microscope was shown as punctated spots on cell margin. The density of Cx43 and eosinophil staining was significantly different between groups of inferior turbinate and nasal polyp (p = 0.01 and 0.03, respectively). Decreased Cx43 expression in the subepithelial fibroblast was correlated with eosinophil infiltration in nasal polyps. Spearman rank order coefficient was equal to -0.43 (p < 0.05). CONCLUSION: This is the first demonstration of decreased Cx43 expression related to eosinophil infiltration. To the best of our knowledge, interleukin-8 may be a link between Cx43 and eosinophil and orchestrating both in developing nasal polyps.     

蛋白激酶C在鼻息肉组织嗜酸性粒细胞浸润增多中的调控作用

目的 :探讨蛋白激酶C(PKC)何种亚型对鼻息肉组织中嗜酸性粒细胞 (EOS)浸润增多具有调控作用。方法 :采用原位杂交法和免疫组织化学MGG染色 ,观察 2 6例鼻息肉组织中PKC几种亚型PKCα、PKCβ1、PKCβ2 、PKCγ的表达 ,分析何种亚型在鼻息肉EOS增多中起调控作用。 结果 :2 6例鼻息肉组织EOS中均有PKC表达 ,且与Bcl 2mRNA及其蛋白成正相关 (r1=0 .0 87 5 ,r2 =0 .0 82 3,P <0 .0 1) ,其中PKCα为强表达 ,而PKCβ1、PKCβ2 为弱表达 ,PKCγ则不表达。结论 :EOS增多主要是由于激活了PKC这一信号传导途径 ,使EOS凋亡受到抑制、生存延长 ,致使其数量增多 ,且主要是PKC亚型中的PKCα在鼻息肉组织EOS增多中起调控作用     

The regulation of PKCalpha in eosinophil infiltration and proliferation in nasal polyps]

OBJECTIVE: To explore the regulation of protein kinase C (PKC) isoform--PKCalpha in eosinophil (EOS) proliferation and infiltration in nasal polyp tissues. METHOD: With the methods of in situ hybridization staining and immunohistochemistry MGG staining, to check out the relationship between PKC and bcl-2/BaxmRNA and associated protein, especially PKC isoform--PKCalpha, PKCbeta1 , PKCbeta2, and PKCgamma did not express at all. RESULT: There were PKC expression in the eosinophils of 26 cases from nasal polyps, and the expression of PKC and Bcl-2 mRNA/their protein in EOS of nasal polyps showed remarkably positive relation (r1 = 0.0875, r2 = 0.0823, P < 0.01), but in PKC isoforms, PKCalpha expression was the strongest, but PKCbeta1 and PKCbeta2 expressed thinner and PKCgamma did not express at all. CONCLUSION: The reason of eosinophil proliferation and infiltration in nasal polyps is that PKC signal transduction pathway was activated, and leaded to inhibition of eosinophil apoptosis, and eosinophil survival was delayed, and eosinophil proliferated and infiltrated, and in PKC family, PKCalpha is main.     

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...     

STAT6在鼻息肉组织中的表达及其与嗜酸粒细胞浸润的关系

目的：研究信号转导和转录激活因子6（STAT6）在鼻息肉组织中的表达及其对嗜酸粒细胞（EOS）浸润聚集的作用,探讨其在鼻息肉发生中的可能作用。方法：选取符合纳入标准的鼻息肉患者手术切除标本（鼻息肉组）30例和同期单纯行鼻中隔偏曲矫正术中切除的下鼻甲组织（对照组）10例。采用免疫组织化学SP法检测2组下鼻甲黏膜中STAT6的表达。应用SPSS13.0软件进行统计学分析。结果：STAT6和EOS在鼻息肉组织中的表达明显高于下鼻甲,差异有统计学意义（P〈0.05）。STAT6阳性细胞主要集中于鼻息肉的上皮细胞、腺体细胞和组织中浸润的炎性细胞的细胞质中。鼻息肉组中STAT6的表达与EOS浸润程度一致（P〈0.01）。结论：STAT6在鼻息肉组织中的高表达及其对EOS浸润聚集的作用,可能与鼻息肉的发生和发展关系密切。     

IL-12和IL-4在鼻息肉表达的意义

目的探讨鼻息肉组织中T辅助细胞1（Th1）和Th2细胞及细胞因子IL 12和IL 4的含量及意义。方法标本取自30例鼻息肉患者的息肉组织和20例行鼻中隔手术患者的正常中鼻甲黏膜（对照组）。新鲜鼻息肉手术标本和正常中鼻甲黏膜制备单细胞悬液,流式细胞术测定Th1和Th2细胞的百分率。用酶联免疫吸附测定（ELISA）法检测IL 12和IL 4在冰冻鼻息肉标本和正常中鼻甲黏膜的含量。结果鼻息肉组织以Th1细胞亚群为主,且其中Th1和Th2细胞亚群显著高于对照组（P<0.05）;鼻息肉组织IL 12、IL 4的水平高于对照组（P<0.05）;Th1细胞百分率与IL 12的水平正相关（P<0.05）,Th2细胞百分率与IL 4的水平正相关（P<0.05）,IL 12的水平与IL 4负相关（P<0.05）。结论鼻息肉中Th细胞在IL 12和IL 4作用下分化为Th1和Th2,二者共同参与鼻息肉的发生发展和免疫应答。     

嗜酸性粒细胞浸润程度与鼻息肉术后疗效的相关性分析

目的探讨嗜酸性粒细胞在鼻息肉中的浸润程度与术后疗效的关系。方法收集眉山市人民医院2014~2017年接受手术治疗的124例鼻息肉（包括Ⅱ、Ⅲ型）患者为研究对象,术后对鼻息肉中嗜酸性粒细胞浸润程度与疗效进行分析。结果术后随访6个月,Ⅱ型1期有效率为100.00％,Ⅱ型2期有效率为88.00％,Ⅱ型3期有效率为100.00％,Ⅲ型有效率为82.61％,Ⅱ型1期和3期有效率均高于Ⅱ2期和Ⅲ型 ,Ⅱ型2期有效率高于Ⅲ型 ,但各组比较差异均无统计学意义（P>0.05）;Ⅰ度嗜酸性粒细胞浸润的鼻息肉手术有效率为100.00％,Ⅱ度有效率为96.43％,Ⅲ度有效率为75.00％, Ⅳ度有效率为0.00％,Ⅰ度有效率均高于Ⅱ、Ⅲ和Ⅳ度（χ2=4.009、11.786、52.000,P<0.05）,Ⅱ、Ⅲ度手术有效率高于Ⅳ度（χ2=19.371、12.000,P<0.05）,Ⅱ与Ⅲ度比较差异无统计学意义（χ2=3.292,P>0.05）;Spearman相关性分析显示,嗜酸性粒细胞浸润程度与术后疗效呈负相关（P<0.05）。结论嗜酸性粒细胞浸润程度与鼻息肉术后疗效相关,嗜酸性粒细胞浸润程度与鼻息肉术后疗效有关,嗜酸性粒细胞浸润程度越深,鼻息肉术后疗效越差,手术联合白三烯受体拮抗剂与鼻喷激素,对难治性慢性鼻-鼻窦炎伴鼻息肉患者有较好的治疗效果。     

Expression of RANTES by IL-1 beta and TNF-alpha stimulated nasal polyp fibroblasts

OBJECTIVE: Accumulation of eosinophils (Eo) is one of the most characteristic feature of nasal polyps. However, the question remains why eosinophils accumulate into the nasal polyp tissue. RANTES (regulated upon activation, normal T cell expressed and presumably secreted) is a recently described chemokine that is said to play a role in the recruitment of eosinophils into inflammatory tissue sites. Fibroblasts are a rich source of cytokines and inflammatory mediators. The objective of this study was to demonstrated the expression of the chemokine RANTES in nasal polyp fibroblasts after stimulation with proinflammatory cytokines like TNF-alpha and IL-1 beta. METHODS: Fibroblast lines were established from human nasal polyp biopsy tissues taken from patients with chronic sinusitis who had no other associated diseases. Cultured nasal polyp fibroblasts were stimulated with TNF-alpha or IL-1 beta at various doses (0.1, 1.0, 1 ng/ml) or for various times (l, 6, 12, 24, 48, 72 h). To detect the RANTES gene expression, RT-PCR was performed. The resulting supernatants were assayed with ELISA for the level of RANTES. RESULTS: We demonstrated that TNF-alpha and IL-1 beta induced the gene expression and protein production of RANTES in nasal polyp fibroblasts. This responsiveness to TNF-alpha and IL-1 beta was time and dose-dependent. CONCLUSION: These findings suggest that nasal polypfibroblasts may also play an important role in the recruitment of Eo through the production of RANTES.     

TNF-alpha and endotoxin increase hypoxia-induced VEGF production by cultured human nasal fibroblasts in synergistic fashion

OBJECTIVE: Vascular endothelial growth factor (VEGF) promotes angiogenesis and is associated with the invasion and metastasis of malignant tumors. It enhances vascular permeability and is expressed in inflammatory nasal as well as middle-ear mucosa. As the mechanism of VEGF induction during chronic inflammation, such as chronic paranasal sinusitis (CPS) remains to be clarified, we studied the factors regulating the production of VEGF in cultured human nasal fibroblasts and discussed the role of VEGF in the pathogenesis of CPS. METHODS: We used ELISA to quantify VEGF levels in paranasal sinus effusions, nasal secretions, and serum from patients with CPS. In addition, we cultured human nasal fibroblasts isolated from nasal polyps of CPS patients and studied the effects of hypoxia, TNF-alpha, and endotoxin on their production of VEGF using ELISA and PCR. RESULTS: The VEGF concentration was significantly higher in paranasal sinus effusions than in nasal secretions and serum. Nasal fibroblasts produced high levels of VEGF, when cultured under hypoxic condition and this production was remarkably enhanced in the presence of TNF-alpha or endotoxin. CONCLUSION: VEGF is locally produced in paranasal sinuses as well as nasal mucosa and its production is increased in patients with CPS. Hypoxia is associated with the production of VEGF by nasal fibroblasts and TNF-alpha and endotoxin may act synergistically to enhance VEGF production in paranasal sinuses under hypoxic condition.     

鼻息肉中嗜酸性粒细胞浸润和活化状况的研究

目的 :观察鼻息肉组织中嗜酸性粒细胞浸润和活化状况 ,探讨嗜酸性粒细胞在鼻息肉发病机制中的作用。方法 :16例鼻息肉组织标本 ,采用组织化学染色 Chromotrope 2R染色法 ,标记鼻息肉组织中的嗜酸性粒细胞 ,结合应用嗜酸性粒细胞阳离子蛋白抗体EG2 的免疫组织化学染色结果 ,观察鼻息肉中嗜酸性粒细胞的浸润和活化状况。结果 :①鼻息肉组织中有较多嗜酸性粒细胞浸润 ,且多处于活化状态 ;②变应性患者鼻息肉组织中EG2 阳性细胞密度、Chromotrope 2R阳性细胞的密度及嗜酸性粒细胞活化比率 ,分别与非变应性患者相比较 ,均无显著性差异 (P >0 .0 5 )。结论 :①Chromotrope 2R特染法结合应用EG2 抗体的免疫组化染色法简便易行 ,适合临床应用观察鼻息肉中嗜酸性粒细胞的浸润和活化状况 ;②活化嗜酸性粒细胞在鼻息肉的发病机制中可能起重要作用。     

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。     

鼻息肉成纤维细胞在常氧与低氧环境下的原代培养及鉴定

目的 探讨鼻息肉成纤维细胞在常氧与低氧环境下的原代培养及鉴定,以备后续研究其生物学特性。方法 取2012至2013年行鼻内镜下鼻息肉切除的息肉组织20例。标本放入4℃无菌的PBS溶液中,在超净台中反复冲洗、浸泡,剪成1～2mm大的组织块后,分成2份转入EP管中,分别用0.1％Ⅰ型胶原酶和1mL的dispaseⅡ酶消化过夜。次日取出,轻轻吹打,进行计数后,加入培养液(DMEM+10％胎牛血清+1×105U/L青霉素+100mg/L链霉素),分别置于常氧、低氧条件下培养(常氧培养以空气作为混合气体,氧体积分数约为20%;低氧培养则以氮气作为混合气体,调节氧体积分数为5%。二者其他培养条件相同,均为37℃、含体积分数为5%的CO2和饱和湿度) 。在倒置相差显微镜下观察细胞的形态、增殖及生长状况、有无污染。用CCK-8比色法检测细胞的增殖,描绘生长曲线。结果 dispaseⅡ酶消化组的细胞数目高于Ⅰ型胶原酶消化组,两组之间差异有统计学意义(P<0.05)。鼻息肉成纤维细胞在常氧及低氧条件下培养的生长曲线无明显差异。结论 dispaseⅡ酶消化的细胞数目较多,是一种较好的获得细胞的方法。在常氧及低氧条件下均可建立稳定、可靠的人鼻黏膜成纤维细胞原代培养模型, 可为鼻息肉的相关研究提供良好的细胞系。     

IL-36在慢性鼻-鼻窦炎伴鼻息肉中的研究进展

白介素-36(IL-36)是IL-1超家族成员(IL-1F)之一,在上皮细胞和特异性免疫细胞中具有生物活性,主要功能包括促进细胞的活化、分泌细胞因子和趋化因子、招募和激活不同的免疫细胞等。近年有学者研究发现,IL-36家族在慢性鼻-鼻窦炎伴鼻息肉(CRSwNP)发病中具有一定的作用。就IL-36的生物学特征及其在慢性鼻-鼻窦炎伴鼻息肉中促进炎症反应、组织重塑、破坏上皮屏障等方面的研究进展进行综述。     

Eosinophilic nasal polyps are a rich source of eotaxin, eotaxin-2 and eotaxin-3

INTRODUCTION: The CC-chemokine eotaxin plays a key role in the pathologic mechanism of tissue eosinophilia in nasal polyposis. In this study, we investigated a possible role of eotaxin-2 and eotaxin-3, the recently discovered members of the eotaxin family. METHODS: Nasal polyps from 24 patients (non allergic/allergic/aspirin-intolerant patients) and turbinate tissue from 8 controls were investigated. Chemokine protein content (eotaxin, eotaxin-2, and -3) of tissue homogenates was measured by ELISA. Paraffin sections of samples were stained to determine the extent of eosinophilia. RESULTS: Protein expression of eotaxin, eotaxin-2 and eotaxin-3 was significantly higher in nasal polyps than in controls. There was a direct correlation between the protein concentrations of all three eotaxins. Further, protein levels of all chemokines were significantly correlated to the amount of eosinophilia. In aspirin-sensitive polyps the number of eosinophils was significantly higher than in the other patient groups and they had significantly higher eotaxin, eotaxin-2, and -3 protein levels than non-allergic and significantly higher amounts of eotaxin-3 compared with allergic patients. CONCLUSIONS: Our findings suggest, that all members of the eotaxin family are involved in the pathogenesis of nasal polyposis. The results are more likely indicative of a complex cooperation between all members of the eotaxin family than of a specific role in the development of eosinophilia and nasal polyposis.