Protein kinase C in eosinophil proliferation and infiltration in nasal polyp tissue]

OBJECTIVE: To explore the significance of protein kinase C (PKC) in eosinophil (EOS) proliferation and infiltration in nasal polyp tissues. METHODS: With the methods of in situ hybridization and immunohistochemistry staining, PKC and apoptosis gene (Bcl-2, Bax) expression were measured between nasal polyps from 26 patients and inferior turbinate mucous membrane tissue (ITMMT) from 20 normal persons. EOS was identified with May-Grünwald-Giemsa (MGG) staining. RESULTS: In EOS, the positive cell expressive rate of Bcl-2 mRNA and its protein were significantly higher in group of nasal polyps than that in group of ITMMT (P < 0.01). Although the positive cell expressive rate of Bax mRNA and its protein were slightly higher in group of nasal polyp tissue than that in group of ITMMT, the difference was not significant (P > 0.05). PKC was expressed in EOS of all 26 cases from nasal polyp tissues, but in 20 cases from ITMMT, only 7 cases showed occasional PKC expression. In both groups, PKC positive cell expression was significantly different, and the expression of PKC and Bcl-2 mRNA as well as their protein in EOS of nasal polyps showed remarkably positive relation (r1 = 0.0875, r2 = 0.0823, P < 0.01). CONCLUSION: PKC expression increasing in EOS of nasal polyp tissues was closely associated with apoptosis inhibiting. So we presumed that EOS inhibiting in nasal polyp tissues was obtained by activating PKC signal transduction pathway.     

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.     

Significance and expression of transforming growth factor beta in human nasal polyp tissue]

OBJECTIVE: To explore the pathogenesis of nasal polyposis and the expression of transforming growth factor beta (TGF-beta) in human inflammatory nasal polyps. METHODS: TGF-beta 1-3 in nasal polyp tissues and inferior turbinate mucosa of twenty-five polyposis patients were detected with immunohistochemistry alkaline phosphatase and anti-alkaline phosphatase (APAAP) method. The inferior turbinate mucosa of eight healthy volunteers were selected as control. Six polyp tissues were estimated with double immunolabeling and Western-blot analysis to compare the characterization of the TGF-beta isoforms expression and the proportion of macrophages and eosinophils in nasal polyp tissues. RESULTS: The expression of TGF-beta 1-3 in nasal polyps was significantly higher than that in nasal mucosa and indetecable in nasal mucosa from healthy volunteers; TGF-beta 1 was the main isoform detected in nasal polyps; TGF-beta positively was accompanied by numerous macrophage and eosinophil infiltration. CONCLUSIONS: TGF-beta mainly TGF-beta 1 is strongly expressed in nasal polyps and its mucosa, where it could be produced by macrophages and eosinophils. TGF-beta could induce modification of epithelium and connective tissue and therefore be involved in the pathogenesis of nasal polyposis.     

Topical budesonide treatment reduces endothelial expression of intercellular adhesion molecules (vascular cell adhesion molecule-1 and P-selectin) and eosinophil infiltration in nasal polyps.

Infiltration of eosinophil granulocytes and endothelial expression of vascular cell adhesion molecule-1 (VCAM-1) and P-selectin was investigated in biopsies of polyps and inferior turbinates from 16 patients with nasal polyps, by the use of immunohistochemical staining and stereological quantification before, during and after topical treatment with budesonide (Rhinocort Turbuhaler). Before glucocorticoid treatment a higher density of eosinophil profiles (p < 0.005), making up a larger proportion of the total cellular infiltrate (p < 0.0005), was found in the polyps compared with the inferior turbinates. Endothelial VCAM-1 expression was higher in polyps than in inferior turbinates (p < 0.005), in contrast with the expression of P-selectin, which was more frequently expressed in the inferior turbinates (p < 0.05). Topical glucocorticoid treatment reduced the density of eosinophil profiles (p < 0.05) and the endothelial expression of VCAM-1 (p < 0.007) and P-selectin (p < 0.02) in polyps. Eosinophil counts and VCAM-1 expression returned to pre-treatment levels 8 weeks after discontinuation of budesonide treatment. The observed reduction in endothelial expression of cellular adhesion molecules may interfere with cellular recruitment in nasal polyps and thus contribute to the known anti-inflammatory effect of glucocorticoid in nasal polyposis.     

鼻息肉中CD34的表达及其与嗜酸粒细胞浸润的关系

目的 探讨CD34在人鼻息肉组织中的表达及其与嗜酸粒细胞浸润的关系。方法 采用HE和免疫组化ElivisionPlus二步法，检测30例正常下鼻甲，60例鼻息肉组织中CD34蛋白的表达情况，并对CD34阻性血管进行微血管（MVD））计数，了解微血管的分布及嗜酸粒细胞的浸润情况。结果 鼻息肉组织中CD34阳性表达广泛，正常下鼻甲组织中CD34呈散在的弱阳性表达；鼻息肉组织中MVD明显高于下鼻甲组织中MVD（P〈0．01），与嗜酸粒细胞的浸润具有正相关性。结论 CD34蛋白有可能介导嗜酸粒细胞的浸润，加重对鼻黏膜的损害。     

Survivin在鼻息肉组织中的表达及与嗜酸性粒细胞浸润相关性的研究

目的研究生存素（Survivin）在鼻息肉组织中的表达及其与嗜酸性粒细胞浸润的相关性,探讨Survivin在鼻息肉发病过程中的作用。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）,用HE染色法观察组织中的嗜酸性粒细胞浸润情况,免疫组化法检测组织中Survivin的表达。结果①HE染色显示鼻息肉组嗜酸性粒细胞浸润显著增加。②免疫组织化学染色显示Survivin免疫阳性细胞数及着色强度均明显高于对照组,图像分析显示,鼻息肉组Survivin的积分光密度（IOD,103/HP）为50.21±6.32,与对照组5.67±0.58相比有统计学意义（P〈0.05）。③Spearman等级相关分析显示,鼻息肉中Survivin表达与嗜酸性粒细胞浸润表达密切相关（r=0.673,P〈0.01）。结论鼻息肉组织中Survivin表达、嗜酸性粒细胞浸润均上调,且二者有协同表达关系。鼻息肉可能是通过凋亡抑制基因Survivin使炎性细胞凋亡受到抑制,从而促进炎性细胞主要是嗜酸性粒细胞浸润生长,导致慢性炎症反应。     

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。     

Detection of activated eosinophils in nasal polyps of an aspirin-induced asthma patient

Aspirin-induced asthma (AIA) is frequently accompanied by nasal polyps. Eosinophil infiltration is a characteristic feature of nasal polyps associated with AIA. Even though steroids are well known to be effective on managing AIA and its nasal polyps, histochemical examinations after steroid therapy and at recurrence, involving eosinophil infiltration of nasal polyps, have been less studied. To know the histochemical effects of steroid treatment on eosinophil accumulation in nasal polyps of AIA and the histochemical feature of a recurring polyp and to detect distributional differences between storage and secreted forms of eosinophil cationic proteins, we carried out immunocytochemical labelling with antibodies against EGI (recognizing resting and activated eosinophils) and EG2 (recognizing only activated eosinophils), and determined eosinophil infiltration in nasal polyps that were obtained before and after steroid treatment, and at recurrence of polyps. A large number of eosinophils in AIA polyps were found before steroid treatment and at recurrence, and they were predominantly composed of activated eosinophils (EG2-positive). In contrast, eosinophil infiltration was rare in polyps obtained immediately after steroid treatment. This finding suggests that eosinophil infiltration may be associated with nasal polyp formation in AIA, and that activation of eosinophils plays an important role in accumulation of eosinophils and polyp formation beginning with the initial stage.     

还氧合酶-2与鼻息肉发病的关系

目的探讨还氧合酶-2(COX-2)在鼻息肉组织中的表达,分析它在鼻息肉发病过程中的作用,为鼻息肉发病机制的研究提供理论依据,并为鼻息肉的药物治疗寻找新的特异性靶点。方法选取34例未应用任何药物的鼻息肉手术组织标本,根据1997年海口会议标准,分为鼻息肉A组（Ⅱ型1、2期鼻窦炎鼻息肉）和鼻息肉B组（Ⅱ型3期及Ⅲ型鼻窦炎鼻息肉）。同时取10例手术患者的下鼻甲游离缘作为正常对照。采用免疫组织化学SP法对鼻息肉和正常下鼻甲黏膜组织中的COX-2进行观测并拍照。采用JD901图像分析软件分析阳性染色面积和染色密度。结果 COX2在正常下鼻甲黏膜组织中无或极少量表达,在鼻息肉组织中均显示高表达（P<0.01）;Ⅲ型鼻息肉组织中的阳性表达面积和密度均高于Ⅱ型鼻息肉组(P<0.05)。结论COX-2在鼻息肉组织中高表达,且与鼻息肉的分型有关,表明其催化产物PGE2参与了鼻息肉的发生发展。     

Interaction between fibronectin and eosinophils in the growth of nasal polyps

OBJECTIVES: The aim of this study was to determine roles of fibronectin and eosinophils in the etiology of nasal polyposis. STUDY DESIGN: We designed a cohort study of cases with nasal polyposis. Sampled nasal polyps were differentiated by their histopathologic characteristics, and compared by size and computed tomography (CT) stage. METHODS: The size of nasal polyps was determined on the basis of the endoscopic findings, and the extent of sinusitis was evaluated by CT staging. Nasal polyp samples were taken from 82 patients during ethmoidectomy and differentiated by morphologic characteristics, infiltration cell types, or fibronectin positivity. Then their sizes and CT stages were compared. In addition, correlation among these histological characteristics was analyzed. RESULTS: Nasal polyps showing edematous morphology, eosinophil infiltration, or fibronectin expression were significantly large in size. Concerning CT stages, only the infiltration cell type showed a significant difference. Significant correlation among edematous morphology, eosinophil infiltration, and fibronectin expression was also recognized. CONCLUSIONS: These findings suggest that interaction between eosinophils and fibronectin may play a role in edema formation, which contributes to the growth of nasal polyps.     

嗜酸性粒细胞与鼻息肉术后复发和预后的初步探讨

目的研究嗜酸性粒细胞与鼻息肉术后复发和预后的关系。方法对51例鼻息肉病人术前鼻息肉组织和复发鼻息肉组织行嗜酸性粒细胞定量测定，并进行比较。结果嗜酸性粒细胞浸润程度越深，鼻息肉越易复发。结论嗜酸性粒细胞浸润程度与鼻息肉复发直接相关，可做为术后鼻息肉是否治愈的判断指标之一。     

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.     

Expression of ecalectin, a novel eosinophil chemoattractant, in nasal polyps

CONCLUSION: Ecalectin, which is produced in the mucosa of nasal polyps, seems to play an important role in the accumulation and activation of eosinophils in nasal polyps, regardless of the presence or absence of atopic predisposition. OBJECTIVE: Ecalectin is a recently discovered eosinophil chemoattractant which elongs to the galectin family. We investigated the expression of ecalectin in nasal polyp tissues associated with various nasal and paranasal diseases in order to clarify the pathogenesis of eosinophilia in nasal polyposis. MATERIAL AND METHODS: Nasal polyps were taken from 56 patients diagnosed as having chronic sinusitis with nasal polyposis. The surgically resected polyps and nasal turbinates were immunohistochemically stained using antibodies against EG2, human mast cell tryptase, CD3 and ecalectin. RESULTS: The number of EG2- and ecalectin-positive cells was significantly higher in nasal polyps than control turbinates. Ecalectin-positive cells were observed in the subepithelial layer, where many EG2-positive cells were present. The number of ecalectin-positive cells correlated significantly with the number of EG2-positive cells in nasal polyps. Many ecalectin mRNA-positive cells were also observed in nasal polyps with an accumulation of EG2-positive cells.     

Eotaxin-1在真菌球型鼻-鼻窦炎合并鼻息肉中的表达与嗜酸性粒细胞浸润的相关性

目的观察真菌球型鼻-鼻窦炎合并鼻息肉的组织病理学特点,检测Eotaxin-1在真菌球型鼻-鼻窦炎合并鼻息肉中的表达及与嗜酸性粒细胞浸润的相关性,探讨Eotaxin-1在真菌球型鼻-鼻窦炎合并鼻息肉发病过程中的作用。方法收集真菌球型鼻-鼻窦炎合并鼻息肉20例和鼻中隔偏曲矫正术患者代偿性肥大的下鼻甲黏膜15例,用HE染色法观察组织中的嗜酸性粒细胞浸润,免疫组织化学法检测组织中Eotaxin-1的表达,比较两组之间的表达差异,分析组织中嗜酸性粒细胞浸润数目与Eotaxin-1表达的相关性。结果①真菌球型鼻-鼻窦炎合并鼻息肉上皮脱落、鳞状上皮化生现象明显,间质为大量疏松结缔组织,其中可见较多的淋巴细胞、嗜酸性粒细胞和巨噬细胞浸润及大量的毛细血管增生,有时可见较多的纤维组织增生,黏膜内未见真菌菌丝和孢子;②Eotaxin-1在下鼻甲黏膜上皮细胞中有少量表达,真菌球型鼻-鼻窦炎合并鼻息肉表达Eotaxin-1明显增多（P＜0.05）,主要由上皮细胞和血管内皮细胞分泌,间质中可见嗜酸性粒细胞和巨噬细胞表达Eotaxin-1;③组织中嗜酸性粒细胞浸润数目和Eotaxin-1的表达明显相关（P＜0.05）。结论真菌球型鼻-鼻窦炎合并鼻息肉组织呈慢性炎症改变,其中可见较多的嗜酸性粒细胞浸润,真菌可能通过介导Eotaxin-1的过量表达,引起嗜酸性粒细胞聚集活化增多,是真菌球型鼻窦炎合并鼻息肉发生的重要因素之一,拮抗Eotaxin-1发挥功能可能对其临床防治有意义。     

HLA-DR and ICAM-1 expression and modulation in epithelial cells from nasal polyps

OBJECTIVE/HYPOTHESIS: Through human leukocyte antigen-DR (HLA-DR) and intercellular adhesion molecule-1 (ICAM-1) expression, nasal epithelial cells could actively participate in the chronic inflammation and eosinophil infiltration observed in nasal polyps. The objective of the study was to evaluate HLA-DR and ICAM-1 expression in polyp epithelium and in a culture model of polyp epithelial cells allowing ciliated and secretory differentiation. STUDY DESIGN: Prospective non-randomized controlled in vitro study. METHODS: The in vitro HLA-DR and ICAM-1 expression was studied under basal conditions or after exposure to interferon-gamma, transforming growth factor-beta1, lipopolysaccharide, dexamethasone, or cetirizine. HLA-DR and ICAM-1 expression was investigated in situ by immunohistochemical staining of polyps and in vitro by immunofluorescent staining of cell cultures. HLA-DR and ICAM-1 were localized in cultured cells by confocal microscopy. Cultured cells expressing HLA-DR and ICAM-1 were quantified by flow cytometry. RESULTS: Both HLA-DR and ICAM-1 showed significant immunostaining of nasal polyp epithelium. In nasal polyp epithelial cell cultures, less than 5% of cells were positive for HLA-DR whereas 40% were positive for ICAM-1 at day 3. In vitro, HLA-DR was mainly located in the cytoplasm and ICAM-1 predominated on the apicolateral cytoplasmic membrane. Comparison of in situ and in vitro results showed that well-differentiated and poorly differentiated cells predominantly expressed HLA-DR and ICAM-1, respectively. Interferon-gamma significantly increased HLA-DR and ICAM-1 expression, whereas transforming growth factor-beta1 significantly decreased HLA-DR expression and lipopolysaccharide significantly increased ICAM-1 expression. CONCLUSION: HLA-DR and ICAM-1 epithelial expression in nasal polyps in situ and in vitro and their in vitro modulation reinforce the active role of epithelial cells in chronic inflammatory diseases of the upper airways.     

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     

催乳素在鼻息肉巨噬细胞中的表达及意义

OBJECTIVE: To detect the expression level and distribution of prolactin (PRL) in nasal polyp and to find out the significance of the mechanism of PRL in the invasion of nasal polyp. METHODS: Twenty-five polyp tissues were obtained from the patients who were subjected to nasal polypectomy in our Department. Inferior turbinate mucosa was used as control obtained from 12 patients with rhinogenous snoring. HE staining was performed for routine histopathologic examination. The expression of PRL in nasal polyps was observed by immunohistochemical staining, and six polyp tissues were estimated through double staining for determining cells which expressed PRL. RESULTS: (1) Positive expression of PRL was significantly stronger (t =4.004, P < 0.01) in 25 nasal polyp tissues (2.05 +/- 0.88) than that in 12 normal inferior turbinate mucosa (0.96 +/- 0.50). Positive expression of macrophage (CD68) was significantly stronger (t = 3.519, P < 0.01) in 25 nasal polyp tissues (1.85 +/- 0.83) than that in 12 normal inferior turbinate mucosa (0.93 +/- 0.52). (2) The PRL expressing cell mainly was the macrophage as demonstrated by double immunohistochemical method. CONCLUSION: PRL derived from macrophages of nasal mucosa may participate in the formation of nasal polyp through its local immune modulation.