变应性鼻炎患者组胺受体表达及与嗜酸性粒细胞浸润相关性分析

目的 通过观察变应性鼻炎患者鼻黏膜中各种组胺受体亚型的表达及分布差异,初步阐明各种组胺受体在变应性鼻炎患者鼻黏膜中的表达,并与嗜酸性粒细胞浸润进行相关性分析,为了解组胺及其受体在变应性鼻炎病理生理学中的作用提供理论基础.方法 选择变应性鼻炎患者共10例,表麻下取下鼻甲黏膜,分别行组胺受体半定量RT-PCR检测以及嗜酸性粒细胞浸润程度评估.结果 半定量RT-PCR检测显示变应性鼻炎患者鼻黏膜中4种组胺受体均有表达,而且HR1表达显著高于HR2、HR3、HR4三种受体(P均<0.01),HR4表达显著高于HR2、HR3(P均<0.01),而HR2,HR3表达无显著差异(P>0.05).HR1,HR4与变应性鼻炎患者鼻黏膜中嗜酸性粒细胞浸润相关,其中HR4相关性相对较高,而HR2,HR3与其浸润无明显相关性.结论 变应性鼻炎患者鼻黏膜中四种组胺受体均有表达,其表达强弱顺序为HR1>HR4>HR2≈HR3.,HR1,HR4与变应性鼻炎患者鼻黏膜中嗜酸性粒细胞浸润相关.     

变应性鼻炎患者鼻黏膜中组胺H4受体的差异性表达

目的:观察新型组胺受体H4在正常人及变应性鼻炎患者鼻腔黏膜中的表达及分布, 初步了解组胺H4受体在变应性鼻炎发病中的作用.方法:选取正常健康者及变应性鼻炎患者各10例,取鼻黏膜分别通过免疫组织化学及RT-PCR方法检测组胺H4受体在蛋白质及转录水平的表达及分布情况并进行对比.结果:变应性鼻炎患者鼻黏膜中组胺H4受体的表达(49676±8541,0.69±0.11)较正常人(25509±6441,0.42±0.08) 显著增加(P<0.05),鼻黏膜结构细胞及免疫细胞均见H4受体表达.结论:H4受体存在于正常人鼻黏膜中,在变应性鼻炎患者鼻黏膜中表达显著增强,提示组胺H4受体可能是介导组胺参与变应性鼻炎发病的重要配体之一.     

Eotaxin在变应性鼻炎中的表达及组胺调控作用的研究

目的：通过变应性鼻炎动物模型了解Eotaxin在变应性鼻炎中的表达以及变态反应炎性递质组胺对Eotaxin表达的影响,以期深入了解变应性鼻炎的发病机制。方法：卵蛋白致敏法建立变应性鼻炎大鼠模型,应用免疫组织化学、RT-PCR及ELISA法,观察组胺及组胺HR1拮抗剂干预后鼻黏膜、血清及鼻腔灌洗液中Eotaxin的表达量或含量变化。结果：变应性鼻炎干预组与变应性鼻炎无干预组相比,鼻腔灌洗液以及鼻黏膜中Eotaxin的表达量较前者显著增加（P〈0.05）,而H1拮抗剂干预组血清、鼻腔灌洗液以及鼻黏膜中Eotaxin增加均显著受到抑制（P〈0.05）。结论：本实验进一步确认了Eotaxin参与了变应性鼻炎发病的理论基础。组胺作为一种变应性鼻炎中的重要炎症递质可以通过影响Eotaxin的表达参与变应性鼻炎的发病过程,且HR1亦参与了这一过程。     

Toll样受体mRNA在慢性鼻窦炎患者鼻黏膜上皮细胞中的表达

目的　探讨Toll样受体 2 (Toll likereceptor 2 ,TLR 2 )及其Toll样受体 4(Toll likereceptor 4,TLR 4)mRNA在慢性鼻窦炎患者和健康成人鼻黏膜上皮细胞中的表达 ,分析TLR 2和TLR 4在鼻黏膜天然免疫中的作用。方法　应用核酸分子原位杂交技术检测 3 0例慢性鼻窦炎患者和 2 1例健康成人鼻黏膜上皮细胞中TLR 2和TLR 4mRNA的表达。结果　TLR 2和TLR 4mRNA在 3 0例慢性鼻窦炎上皮细胞中均有表达 ,且分别与对照组间差异有显著性意义 (t =8 60 5,P <0 0 0 0 5,t =9 0 50 ,P <0 0 0 0 5)。结论　鼻黏膜上皮细胞中可以产生TLR 2和TLR 4,表明鼻黏膜上皮不仅是单纯的物理屏障 ,更可以通过Toll样受体对病原微生物进行识别 ,以直接启动有效的抗感染机制。     

抗组胺药在变应性鼻炎中的作用机制及应用进展

变应性鼻炎(AR)是一种鼻黏膜的慢性非感染性炎症,主要是由机体接触致敏变应原后诱发的由特异性免疫球蛋白E(IgE)介导的、多种炎性细胞(如肥大细胞、嗜碱性粒细胞等)参与的鼻黏膜的高敏反应.组胺在AR发病中发挥重要作用,是引起鼻痒、喷嚏、流涕等临床症状的核心介质.抗组胺药物通常通过拮抗组胺相关受体来阻断组胺与受体结合,从...     

组胺受体及抗组胺药的分类及其在变应性鼻炎中的应用

组胺是体内一种重要的化学介质,可通过活化H1、H2、H3、H4等类型受体参与多种病理生理过程.在变应性鼻炎(AR)的速发相中,肥大细胞脱颗粒释放组胺等炎性介质,组胺与鼻黏膜中H1受体结合触发喷嚏、鼻痒和流涕等症状,应用H1受体拮抗剂(H1RAs)可显著缓解上述症状.新近研究发现鼻黏膜中除H1受体外还存在H2、H3和H4...     

趋化因子及其受体在变应性鼻炎中基因表达的临床研究

目的：探讨趋化因子及其受体在变应性鼻炎发病机制中的作用。方法：采用基因芯片技术在芯片上点阵30多个趋化因子及其受体的cDNA．对4例变应性鼻炎鼻黏膜及止常人鼻黏膜组织进行基因芯片检测．观察趋化因子及其受体的表达情况。结果：在4例变应性鼻炎鼻黏膜组织标本中．存在eotaxin-1(CCL11)．eotaxin-2(CCL24)．MCP-3(CCL7)。MCP-4(CCL13)。RANTES(CCL5)等趋化因子和CCR2、CCR3、CCR4、CCR5等趋化因子受体基因的差异表达．其中大多数趋化因子及受体倾向Th2优势。结论：变应性鼻炎存在Th免疫失衡,趋化因子及其受体特别是Th2优势的趋化因子及受体在变应性鼻炎发病机制中发挥重要作用．这为变应性鼻炎的诊治提供新的着眼点。     

p38丝裂原激活蛋白激酶通路对变应性鼻炎中黏蛋白5AC的信号转导机制初探

目的:探讨组胺诱导的离体鼻黏膜组织中,p38丝裂原激活蛋白激酶(p38MAPK)信号通路及环氧合酶-2(COX-2)对黏蛋白5AC(MUC5AC)表达的影响,以期阐明变应性鼻炎(AR)中黏液过度分泌的病理学机制。方法:应用Western blot技术检测不同浓度梯度的组胺诱导及p38MAPK特异性抑制剂SB203580或COX-2特异性抑制剂NS-398干预前后,体外培养的鼻黏膜组织中p38MAPK、COX-2和MUC5AC的蛋白质表达变化规律。结果:p38MAPK、COX-2和MUC5AC在体外培养的健康鼻黏膜组织中呈微弱表达。组胺呈浓度依赖性诱导鼻黏膜组织中p38MAPK、COX-2和MUC5AC蛋白质表达增加;以不同浓度梯度的NS-398阻断COX-2后,观察到NS-398呈浓度依赖性减弱组胺对MUC5AC蛋白质表达的诱导,对p38MAPK表达无明显影响。以不同浓度梯度的SB203580阻断p38MAPK后,观察到SB203580呈浓度依赖性减弱组胺对COX-2和MUC5AC蛋白质表达的诱导。培养液中单独加入SB203580或NS-398对MUC5AC的蛋白质表达无明显影响。结论:组胺通过诱导p38MA...     

Toll样受体在真菌性鼻及鼻窦炎的表达及意义

目的研究Toll样受体2(Toll—like receptor 2,TLR2)和Toll样受体4(Toll—like receptor 4,TLR4)在真菌性鼻及鼻窦炎(fungal rhinosinusitis,FRS)患者鼻黏膜中的表达,并探讨其在FRS发病中的可能作用。方法采用逆转录-聚合酶链反应和免疫组化SP法分别检测24例FRS、24例慢性鼻及鼻窦炎(chronic rhinosinusitis,CRS)和15例鼻中隔偏曲手术患者正常下鼻甲鼻黏膜中TLR2和TLR4 mRNA和蛋白的表达水平。结果TLR2和TLR4 mRNA在FRS组的表达水平(0.208±0.063和0.1 59±0.059)明显低于CRS组(0.505±0.097和0.406±0.102)和鼻中隔偏曲患者组(0.327±0.047和0.232±0.088,P均<0.05)。FRS组TLR2和TLR4蛋白的阳性表达率明显低于CRS组和鼻中隔偏曲组(P<0.05)。结论TLR2和TLR4在FRS中表达下调,提示Toll样受体介导的天然免疫在鼻黏膜抗真菌防御反应中作用下降,是引起鼻黏膜真菌清除率下降和真菌易感性增加的原因之一。     

组胺H3受体激动剂IMETIT对豚鼠变应性鼻炎作用的初步探讨

目的:探索组胺H3受体激动剂(H3R激动剂)IMETIT对豚鼠变应性鼻炎(AR)模型的治疗作用和对P物质(SP)及其受体(SP-R)mRNA 表达的影响.方法:以OVA为致敏原,用豚鼠AR模型,观察症状及鼻黏膜病理状况以研究IMETIT对豚鼠AR 的治疗作用;免疫组织化学检测鼻中隔黏膜中SP含量,RT-PCR测定鼻中隔黏膜中SP-R mRNA 表达以研究IMETIT对SP/SP-R分泌及表达的影响.结果:组胺H3R激动剂IMETIT能有效改善豚鼠AR喷嚏(P＜0.01)、鼻痒(P＜0.01)、鼻塞(P＜0.05),减轻鼻黏膜病理性改变,调低SP/SP-R mRNA分泌及表达(P＜0.05或P＜0.01).结论:组胺H3R激动剂IMETIT能有效缓解豚鼠AR的症状,其作用机制与调低SP/SP-R 分泌及表达有关.     

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.     

Survivin及Caspase-3在鼻腔鼻窦内翻性乳头状瘤及鼻腔鳞状细胞癌中的表达及其意义

目的探讨细胞凋亡抑制蛋白Survivin及半胱氨酸天冬氨酸蛋白酶3(Caspase-3)在鼻腔鼻窦内翻性乳头状瘤(sinonasal inverted papillomas,SNIP)及鼻腔鳞状细胞癌(nasal squamous cell carcinoma,NSCC)中的表达及其意义。方法采用免疫组化Envision二步法检测37例SNIP、19例NSCC及20例下鼻甲黏膜组织标本中的Survivin及Caspase-3的表达。结果 SNIP组和NSCC组中Survivin的表达率分别为67.6%(25/37)、89.5%(17/19),均显著高于下鼻甲黏膜组的表达率0(0/20,P<0.01)。NSCC组中Caspase-3的表达率为26.3%(5/19),显著低于下鼻甲黏膜组的表达率100.0%(20/20,P<0.01),SNIP组中Caspase-3的表达率为43.2%(16/37),低于下鼻甲黏膜组,但两者之间的差异无显著性(P>0.05)。Survivin和Caspase-3在SNIP和NSCC中的表达呈负相关(P<0.01)。结论 Survivin可能在SNIP和NSCC发生和发展过程中起着重要的促进作用,且与Caspase-3呈负相关,Survivin可能成为SNIP和NSCC基因治疗的新靶点。     

糖皮质激素受体-α表达下调与鼻息肉相关性的研究

目的 检测糖皮质激素受体α(glucocorticoidreceptor-α,GR-α) mRNA在鼻息肉组织及健康人群鼻腔下鼻甲黏膜、窦口鼻道复合体(ostiomeatal complex,OMC)区黏膜的定量表达,并探讨其在鼻息肉发生发展中可能存在的的机制.方法 采用荧光定量逆转录-聚合酶链反应(FQ-RT-PCR...     

Immunohistochemical localization of histamine receptor subtypes in human inferior turbinates

Histamine is an important chemical mediator in allergic rhinitis and plays an important role in eliciting the nasal symptoms of the disorder. However, the immunohistochemical localization of histamine receptor subtypes (H1R, H2R, H3R, and H4R) in human nasal mucosa is unknown. There are also no prior studies of H3R and H4R in human nasal mucosa. The objective of this study was to examine the distribution of histamine receptor subtypes in the human inferior turbinates by an immunohistochemical method. H1R was localized primarily in the epithelium, vessels, and nerves. H2R was localized primarily in the epithelium and the glands. H3R and H4R were clearly distributed on the nerves. In addition, H1R, H3R, and H4R were clearly localized on the same nerves. This result indicates that H1R, H3R, and H4R adjoin and regulate each other in the same nerves. All histamine receptor subtypes may play some role in patients with allergic rhinitis.     

VEGF在伴鼻息肉的慢性鼻-鼻窦炎中的表达及克拉霉素对其的调节作用

目的：探讨血管内皮生长因子（VEGF）在伴鼻息肉的慢性鼻-鼻窦炎（CRSwNP）患者鼻黏膜中的表达及克拉霉素对其的调控作用。方法：收集16例CRSwNP组织及21例单纯鼻中隔偏曲患者下鼻甲黏膜,采用实时定量RT-PCR方法检测样本中VEGF的表达。另外收集10例伴鼻息肉的慢性鼻窦炎鼻息肉组织及5例单纯鼻中隔偏曲患者下鼻甲黏膜,采用体外组织块培养及ELISA方法,观察克拉霉素对VEGF表达的调节作用。结果：①VEGF mRNA在CRSwNP组的表达较对照组增高,差异有统计学意义（P〈0.01）;②经克拉霉素刺激前后比较,CRSwNP组织中VEGF蛋白的表达量下降,差异有统计学意义（P〈0.05）。结论：CRSwNP组织中VEGF的表达显著升高,推测VEGF在鼻息肉的发病机制中具有极其重要的作用。克拉霉素可能通过抑制VEGF的表达而达到治疗慢性鼻-鼻窦炎的作用。     

辛芩颗粒联合H1受体拮抗剂治疗变应性鼻炎实验研究

目的探索辛芩颗粒联合H1受体拮抗剂氯雷他定对豚鼠变应性鼻炎（allergic rhinitis,AR）模型的治疗作用和对P物质（substance P,SP）及其受体（substance P receptor,SP-R）mRNA表达的影响。方法36只清洁级豚鼠随机分为4组,每组9只,分别为AR模型无干预组（A组）、辛芩颗粒干预组（B组）、H1受体拮抗剂氯雷他定于预组（C组）、辛芩颗粒联合氯雷他定干预组（D组）。各组分别给予不同药物干预后进行喷嚏、抓鼻、呼吸频率的行为学观察,同时应用免疫组化检测鼻中隔黏膜中SP分泌,逆转录聚合酶链反应测定鼻中隔黏膜中SP-R mRNA表达。结果各组间在喷嚏、抓鼻、呼吸频率、SP细胞计数及SP—R mRNA的相对表达量比较均有统计学意义（P值均〈0．05）。实验分析显示,辛芩颗粒联合氯雷他定对上述影响具有协同作用（P值均〈0．05）。结论辛芩颗粒联合氯雷他定在缓解豚鼠AR的症状,减少鼻黏膜中SP阳性细胞数量以及降低SP—R mRNA的表达优于单独用药组,并且具有协同作用。     

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.     

TLR2和TLR4在慢性鼻窦炎鼻息肉中的表达及临床意义

目的检测Toll样受体2(toll-like receptor 2,TLR2)和Toll样受体4(toll-like receptor 4,TLR4)蛋白在慢性鼻窦炎鼻息肉黏膜组织中的表达,并探讨其在鼻窦炎鼻息肉发病机制中的作用。方法采用免疫组织化学技术检测62例鼻窦炎鼻息肉黏膜组织中TLR2和TLR4蛋白的表达和分布,同期选取5例正常筛窦黏膜组织进行对照。按照1997年海口制定的"慢性鼻窦炎鼻息肉临床分型分期及鼻内镜下鼻窦手术疗效评定标准"将其分组,比较各组间TLR2和TLR4表达程度的差异。结果①TLR2、TLR4主要表达在鼻腔黏膜上皮细胞、固有层炎性细胞及黏膜下层腺体的胞浆和胞膜上。②慢性鼻窦炎鼻息肉各型各期均较对照组高;并且随着分型的升高,TLR2、TLR4的表达量也随之增加,差异具有统计学意义(P<0.05)。结论 TLR2和TLR4在鼻窦炎鼻息肉中的表达可能与鼻窦炎鼻息肉的发病机制存在着一定的相互关联,TLR在该机制中可能扮演重要角色。     

Thresholds in nasal histamine challenge in patients with allergic rhinitis, patients with hyperreflectory rhinopathy, and healthy volunteers

BACKGROUND: Characteristic symptoms of hyperreflectory rhinopathy include recurrent sneezing, nasal obstruction, and nasal secretion without an allergic background. The diagnosis can only be made if all differential diagnoses have been excluded. So far no clinical test has been established to reliably diagnose hyperreactivity of the nasal mucosa. The present study aimed to investigate whether nasal provocation with histamine allows identification of patients with hyperreflectory rhinopathy. MATERIALS AND METHODS: One-sided nasal challenge with histamine was applied to 13 patients with allergic rhinitis, 13 patients with hyperreflectory rhinitis, and 12 healthy volunteers. Histamine concentrations used were 0.25, 0.5, 1.0, 2.0, 4.0, 8.0, and 16.0 mg/mL. Test results were quantified using a symptom score (positive at values above 3) and active anterior rhinomanometry (positive at a reduction of airflow of 40% or more in comparison to challenge with solvent). RESULTS: While there was a significant difference between controls and patients with allergic rhinitis or hyperreflectory rhinopathy, respectively, no significant difference was observed between the two groups of patients. Results indicated that one-sided nasal provocation with histamine at a concentration of 1 mg/mL is sufficient to separate healthy subjects from patients with hyperreactivity of the nasal mucosa. In terms of the differentiation between subjects with hyperreactivity of the nasal mucosa and healthy controls, the sensitivity of one-sided nasal histamine provocation with 1 mg/mL was found to be 100%; its specificity was 83% if it was evaluated by rhinomanometry and symptom score. CONCLUSION: The present results indicate that one-sided nasal challenge with histamine at a concentration of 1 mg/mL is sufficient to separate healthy subjects from patients with hyperreactivity of the nasal mucosa. However, the test does not differentiate between patients with allergic rhinitis and patients with hyperreflectory rhinitis.     

Expression of TLR2 and TLR4 messenger RNA in the epithelial cells of the nasal airway

BACKGROUND: Epithelium of nasal mucosa is the first line of defense against invading pathogens. This study investigated the expression of Toll-like receptor (TLR) 2 and TLR4 in epithelial cells of nasal mucosa and understood the role of TLRs in the innate immunity of nasal mucosa. METHODS: Human nasal epithelial cells were obtained by scraping the middle one-third of inferior turbinates from 30 patients with chronic rhinosinusitis and 20 healthy adult volunteers. The epithelial cells are made into smears. In situ hybridization was performed for TLR2 and TLR4 messenger RNA (mRNA). RESULTS: TLR2 and TLR4 mRNA were expressed in the nasal epithelial cells. The expression of the two genes was significantly higher in the chronic rhinosinusitis group than in the normal control (TLR2, t = 8.605, p < 0.0005; TLR4, t = 9.050, p < 0.0005). CONCLUSION: This study is the first to establish the presence of both TLR2 and TLR4 mRNA on epithelial cells of nasal mucosa, and their expression can be up-regulated in infectious conditions. These results show that TLR2 and TLR4 may play a important role in local host defense of nasal mucosa.