一氧化氮合酶在鼻息肉中的表达

一氧化氮合酶(nitric oxide synthase，NOS)有3种亚型^[1]，分别为神经元型一氧化氮合酶(neuronic nitric oxide synthase，nNOS)、内皮型一氧化氮合酶(endothelial nitric oxide synthase，eNOS)和诱导型一氧化氮合酶(inducible nitric oxide synthase，iNOS)。NOS可能在鼻炎和鼻息肉形成中起着重要作用。为了解NOS在鼻息肉组织中分布情况     

变应性鼻炎动物模型血一氧化氮和鼻黏膜一氧化氮合酶的动态检测

目的探讨在变应性鼻炎（allergic rhinitis,AR）发病及应用抗过敏药物治疗过程的不同阶段,变应性鼻炎动物模型血一氧化氮（Nitric Oxide,NO）水平和鼻黏膜一氧化氮合酶（Nitric-Oxide Synthase,NOS）表达的动态变化。方法选用健康SD大鼠50只,将其随机分为5组：正常指标组、实验1组、实验2组、实验3组、治疗组。在不同阶段分别取血和鼻黏膜;检测血浆中NO含量及鼻黏膜匀浆液NOS含量。结果实验1组和实验3组动物的血浆NO及鼻黏膜匀浆上清液NOS含量均显著升高,实验2组的NO及NOS含量均无显著变化;治疗组NO含量降至正常指标组水平,NOS亦有部分下降。血浆NO与鼻黏膜NOS含量之间存在显著的正相关关系（r=0．770,P=0．013）。结论血浆NO及鼻黏膜NOS水平在AR造模的不同阶段有明显的波动性变化,常规抗过敏药物能有效抑制患病机体NO的产生。血NO含量的变化能较好地反应鼻黏膜NOS活性的改变。     

丹参对顺铂耳蜗毒性的防护作用

目前有研究发现一氧化氮(nitric oxide,NO)与顺铂导致的内耳毒性关系密切,增加的NO可能参与了顺铂的耳毒性作用。近来又有文献报道,丹参salvia miltiorrhiza)可以通过降低一氧化氮合酶(nitric oxide synthase,NOS)活性以降低庆大霉素耳毒性。但是,丹参能否降低顺铂耳毒性,是否通过影响NOS表达来实现,目前尚未见报道。本研究拟采用顺铂内耳中毒的动物模型,通过免疫组织化学、图像分析、听性脑干反应测定、电镜观察等方法,来探讨顺铂耳蜗毒性的机制,以期寻求防治顺铂耳蜗毒性损伤的有效药物。     

一氧化氮在鼻部变态反应疾病中的细胞机制研究进展

随着对变应性鼻炎、鼻息肉等鼻部变态反应性疾病的不断深入研究 ,许多新的机制不断被发现。如嗜酸性粒细胞 ( EOS)在疾病中作用的重新认识〔1〕,Th1 以及 Th2 细胞平衡关系在疾病中的作用〔2〕等 ,使人们对鼻部变态反应有了更深入的理解。一氧化氮 ( NO)是近年来发现的一种广泛存在于人体内的小分子活性物质。其首先作为一种血管内皮的舒张因子于 1 980年被 Furchgott等发现。NO在体内主要由左旋精氨酸 ( L-Arg)在一氧化氮合成酶 ( NOS)的作用下脱掉末端胍基氮原子而合成〔3〕。NOS分为神经元型 ( n NOS或 NOS1 )、诱导型 ( i NO…     

鼻用糖皮质激素对变应性鼻炎和非变应性鼻炎患者呼吸道一氧化氮浓度的影响

目的　评估鼻用皮质类固醇激素对常年性变应性鼻炎及非变应性鼻炎患者鼻呼吸道一氧化氮（nitric oxide,NO）浓度的影响。方法　根据鼻内镜检查和鼻窦CT扫描排除鼻息肉及鼻窦受累患者,以皮肤点刺试验了解患者的特应性状态,并行鼻分泌物嗜酸性粒细胞检查。共选择收集23例常年性变应性鼻炎患者,17例非变应性鼻炎伴嗜酸性粒细胞增多症（nonallergic rhinitis with eosinophpilia,NARES）患者,20例正常人作为对照组。采集一般病史,视觉模拟量表评分法评估患者鼻部症状的严重程度,并填写鼻-结膜炎相关生活质量问卷,采用NIOX测量鼻呼出气。以布地奈德鼻喷雾剂（64 μg/喷/鼻）喷鼻治疗,2次/天,持续治疗4周,并于2周和4周时复查,再次评估症状、生活质量及鼻腔NO水平。结果　变应性鼻炎患者鼻NO水平明显高于NARES患者[（1053±137）ppb vs（741±65）ppb,P <0.001）];对比正常对照组[（838±79）ppb）],两组患者鼻NO水平较之分别表现为升高和降低,且差异具有统计学意义（P 均<0.001）。经鼻用布地奈德鼻喷雾剂治疗2～4周后,变应性鼻炎患者表现为鼻NO浓度降低,而NARES患者表现为鼻NO升高,且同组治疗前、后差异有统计学意义（P <0.001,P <0.05）。治疗2～4周之后,变应性鼻炎及NARES患者鼻NO水平持续向对照组接近,同时,患者的鼻部症状评分及生活质量评分明显改善。结论　变应性鼻炎和非变应性鼻 炎患者分别表现为NO浓度升高和降低;经鼻用糖皮质激素喷雾治疗后,两组患者鼻腔NO浓度向正常人水平回归。鼻腔NO水平改变可以作鼻炎患者炎症控制的指标。     

氧自由基与分泌性中耳炎

分泌性中耳炎(secretory otitis media，SOM)是以鼓室积液及听力下降为主要特征的中耳非化脓性炎性疾病。关于SOM的发病机理至今未明确，其中以各种原因所致咽鼓管功能障碍最受重视。关于一氧化氮(nitric oxide，NO)、一氧化氮合酶(nitric oxide synthase，NOS)及氧自由基(oxygen free radical，OFR)在SOM中作用的研究较少，本文就此做简要概述。     

头颈部鳞状细胞癌中诱生型一氧化氮合酶的表达及其临床意义

本研究采用硝酸还原酶法测定血清一氧化氮（nitric oxibe,NO)水平，采用逆转录聚合酶链反应（revevse transriptase polymerase chain reaction,PTPCR)法检测组织中诱生型一氧化氮合酶（inducible NOS,iNOS)mRNA表达，探讨其在头颈部鳞状细胞癌（简称鳞癌）发病机制中的作用。     

实验性微栓塞缺血豚鼠耳蜗cNOS的表达变化

目的 探究一氧化氮（nitricoxide，NO）在一过性微栓塞耳蜗缺血性损伤中的作用。方法 20只豚鼠随机分为实验组和对照组各10只，实验组动物通过磁微粒栓塞造成实验性耳蜗缺血模型；以扫描电镜观察缺血耳蜗听纤毛变化；以免疫组织化学方法观察原生型一氧化氮合酶（constitutive nitric oxide synthase，cNOS）在两组耳蜗的表达变化。结果 实验性耳蜗缺血造成内外毛细胞听纤毛散在的粘结、倒伏或缺失，内毛细胞听纤毛病变明显；内皮型一氧化氮合酶（endothelial NOS，eNOS）表达分布于蜗轴及内侧螺旋板内神经纤维、螺旋神经节、内侧螺旋缘、Corti器细胞及血管纹／螺旋韧带区；神经型一氧化氮合酶（neuron NOS，nNOS）主要分布于蜗轴及内侧螺旋板内神经纤维、螺旋神经节细胞，在Corti器细胞、血管纹细胞及内侧螺旋缘上皮细胞有较弱的表达；实验性缺血组豚鼠耳蜗与正常对照组耳蜗原生型一氧化氮合酶的表达无差异。结论 微栓塞耳蜗缺血引起散在听毛细胞损伤；两种原生型NOS在耳蜗固有表达，分布有交叉，功能存在相互代偿，但未发现因微栓塞缺血而引起变化。     

黏蛋白在鼻息肉及变应性鼻炎中的基因表达水平的研究

目的：探讨5种人黏蛋白基因（MUC2、MUC5AC、MUC5B、MUC18、MUC19）在鼻息肉和变应性鼻炎中的基因表达水平及临床意义。方法：采用RT-PCR及实时荧光定量RT-PCR,检测35例鼻息肉、18例变应性鼻炎患者下鼻甲及18例正常人黏蛋白基因（MUCS）在下鼻甲黏膜上皮的基因表达水平。结果：RT-PCR及实时荧光定量RT-PCR的结果表明,MUC5AC、MUC5B在鼻息肉组和变应性鼻炎组中的表达高于对照组（P〈0.05）,鼻息肉组与变应性鼻炎组及对照组间表达差异均无统计学意义（P〉0.05）。MUC2、MUC18在3组中的表达差异无统计学意义（P〉0.05）。MUC19在变应性鼻炎组中的表达高于鼻息肉组和对照组（P〈0.05或P〈0.01）。结论：MUC5AC和MUC5B在鼻息肉、变应性鼻炎的黏液过度分泌过程中具有重要意义;MUC19在变应性鼻炎的黏液过度分泌过程中也有重要意义,但是对鼻息肉患者的黏液分泌没有明显作用;未发现MUC2、MUC18在变应性鼻炎的黏液过度分泌过程中起明显作用。     

呼出气一氧化氮与变应性鼻炎的相关性分析

目的　探讨呼出气一氧化氮（f r a c t io n a l exhaled nitric oxide,FeNO）与变应性鼻炎的相关性。方法　研究66例变应性鼻炎患者视觉模拟量表（visual analogue scale,VAS）评分在治疗前、后与FeNO的相关性。结果　经治疗后FeNO明显下降（t =4.25,P =0.0000）,下降比例为（14.56±27.23）%,VAS评分明显下降（t =3.34,P =0.0008）,下降比例为（8.68±53.17）%。相关性结果分析,治疗前、后变应性鼻炎患者FeNO与VAS评分具有显著相关性（r =0.43,P =0.0003;r =0.45,P =0.0002）。治疗后变应性鼻炎患者FeNO与VAS评分下降比例具有显著相关性（r =0.25,P =0.0437)。结论　FeNO可以对变应性鼻炎的治疗疗效提供客观的数据依据。     

Expression of nitric oxide synthases in nasal mucosa from a mouse model of allergic rhinitis

Nitric oxide (NO), which is produced by nitric oxide synthase (NOS), has been recently identified as a multifunctional mediator. As for nasal tissue, however, the distribution and expression patterns of 3 isoforms of NOS, including neuronal NOS (nNOS, type I NOS), inducible NOS (iNOS, type II NOS), and endothelial NOS (eNOS, type III NOS), are still unclear. To evaluate the function of NO in the pathophysiology of nasal allergy, we investigated the distribution of NOSs in the nasal mucosa of C57BL/6 mice with allergic rhinitis to the house dust mite, Dermatophagoides farinae. Immunoreactivity to each isoform of NOS was immunohistochemically observed. In the allergic nasal mucosa, many eosinophils had infiltrated. Immunoreactivity to NOS types I and III was localized to the surface epithelial and vascular endothelial cells in both allergic and control groups without a statistically significant difference. In contrast, the type II NOS immunoreactivity was weak in normal mice and increased after allergic sensitization. The type II NOS expression of the surface epithelial and vascular endothelial cells was significantly elevated in the allergic group as compared with the control group. These findings suggest that a large amount of NO may be produced in the nasal mucosa of mice by type II NOS after allergic sensitization and that type II NOS may play an important role in the pathogenesis of allergic rhinitis.     

Increased expression of inducible nitric oxide synthase in nasal mucosae of guinea pigs with induced allergic rhinitis

BACKGROUND: Nitric oxide (NO) is produced by the action of NO synthase (NOS) isoforms and is considered an important mediator of inflammatory response including airways. In this study, the changes in the expression levels of NOS isoforms in nasal mucosae were determined in a guinea pig model of allergic rhinitis. METHODS: An allergic rhinitis model was prepared in guinea pigs by repeated challenge with aerosolized dinitrophenylated ovalbumin antigen. Twenty-four hours after the last antigen challenge, the expression levels of NOS isoforms in nasal mucosae were determined by immunoblottings. Changes in the isometrical tension of isolated mucosal tissues of nasal septa induced by histamine were measured also. RESULTS: Although the expression levels of endothelial NOS (eNOS) and neuronal NOS (nNOS) in nasal mucosae were not affected by the repeated antigen exposure, the inducible NOS (iNOS) level was markedly and significantly increased in the challenged animals. In isolated nasal mucosal tissues, histamine induced a concentration-dependent relaxation, which was sensitive to an H1-receptor antagonist, mepyramine, and an NOS inhibitor, L-NMMA. No significant change in the histamine responsiveness was observed between the sensitized control and repeatedly antigen-challenged groups. CONCLUSION: The expression of three isoforms of NOS, including eNOS, nNOS, and iNOS, was presented in guinea pig nasal mucosa. A marked increase in iNOS expression in the repeatedly antigen-challenged animals suggests an important role of iNOS in the pathogenesis of allergic rhinitis. However, the pathophysiological role(s) of NO generated by iNOS in nasal allergy is still unclear.     

Tumor necrosis factor-alpha and interferon-gamma modulate in vitro expression of nitric oxide synthase in human nasal epithelial cells

BACKGROUND: Interest in the physiological, pathological and therapeutic implications of nitric oxide (NO) have grown exponentially, with human nasal cavity and paranasal sinuses considered a dominant source of NO, indicating that this molecule possesses the diversity of biological effects in the regulation of airway clearance and nonspecific cellular immunity. We previously observed differences in NO synthase (NOS) isoform constitutively expressed in nasal epithelial cells (NECs) from allergic and normal subjects. OBJECTIVES: We extended the previous work to determine whether in vitro stimulation with proinflammatory cytokines influences levels of different NOS isoform expression. METHODS: Nasal epithelial cells were sampled from the inferior turbinate in a group of 16 healthy normal controls and 11 patients with perennial allergic rhinitis against house dust (HD) mite antigen. 1 x 10(5) cells were incubated in conditioned medium for 24 hours. Human recombinant interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), or both (cytomix) were added to a final concentration of 10 ng/ml. Cells were then fixed with 4% paraformaldehyde and processed for fluorescence immunocytochemistry. Immunoreactivity for 2 NOS isoforms, inducible NOS (iNOS) and endothelial NOS (eNOS), was studied by laser scanning confocal microscopy and fluorescence intensity was assessed quantitatively. RESULTS: We observed constitutive eNOS expression in epithelial cells of all subjects. Different treatments with cytokines did not affect eNOS expression. Cytokine treatment, however, significantly augmented iNOS expression in the control group. The average increase induced with IFN-gamma, TNF-alpha, and cytomix was 1.8, 2.33, and 2.31-fold. Nasal epithelial cells in the HD group showed elevated steady-state iNOS expression even in untreated. Cytokine treatment did not affect the degree of iNOS expression in this group. CONCLUSION: These results confirm our previous findings that nasal epithelial cells in patients with allergic rhinitis produce higher levels of NO through the concomitant expression of different NOS isoforms. We also demonstrated that nasal epithelial cells have the potential to express iNOS protein spontaneously or upon stimulation with inflammatory cytokines, such as IFN-gamma and TNF-alpha. Because the high level of exhaled NO is considered a potential marker of allergic airway inflammation, preserving the iNOS gene from its unregulated induction may be important for maintenance of nasal homeostasis and may offer a tool for therapeutic intervention.     

Localisation of heme oxygenase isoforms in allergic human nasal mucosa

Carbon monoxide (CO) is an endogenously produced gas mediator produced by heme oxygenase (HO). Like nitric oxide (NO), CO is produced in the nasal mucosa. Given that induced NO synthase (iNOS) expression in nasal mucosa has been found to be up-regulated in allergic rhinitis, the current study investigated the expression of HO isoforms in allergic human nasal mucosa. Immunohistochemical staining for type 1 and 2 HO isoforms were carried out in nasal inferior turbinate mucosa from six patients with persistent allergic rhinitis, and compared with six control patients without nasal allergy. Focal and weak expression of HO-1 was observed in seromucous glands, with no difference between allergic and control specimens. Vascular endothelium, erythrocytes, smooth muscle and inflammatory cells (except macrophages) in the allergic group exhibited stronger HO-1 immunoreactivity compared to the control. Minimal expression was found in the respiratory epithelium in either group. Intravascular HO-1 expression was found in the allergic mucosa only. Intense HO-2 immunoreactivity was observed in the respiratory epithelium, vascular endothelium and seromucous glands in both allergic and control groups with no differences in intensity. In conclusion, unlike iNOS, HO-1 is minimally expressed in the nasal respiratory epithelium of either group. However, our findings suggest that it may be involved in the inflammatory process of allergic rhinitis at the submucosal level.     

Increased expression of inducible nitric oxide synthase in nasal epithelial cells in patients with allergic rhinitis

OBJECTIVES: Although ciliated epithelial cells of human nose and paranasal sinuses have recently been reported to be the major source of locally detected nitric oxide (NO), changes to the NO production by these cells and their functional roles remain uncertain in relation to allergic rhinitis. The objective of this study is to investigate differences in the ability of induction of nitric oxide synthase (NOS) isoforms by nasal epithelial cells. STUDY DESIGN: Epithelial cells of the inferior turbinate taken from 12 normal subjects and 12 allergic patients against house dust mite were used. Samples from the house dust group were taken both before and after antigen provocation. METHODS: Immunoreactivity for two NOS isoforms, inducible NOS (iNOS) and endothelial NOS (eNOS), was examined by the laser scanning confocal microscope. The labeled cells were processed into digital images, and the fluorescence intensity was assessed quantitatively. RESULTS: The degree of iNOS expression of the epithelial cells was significantly elevated in the house dust group compared with that of the control group. The expression appeared identical both before and after antigen provocation in the house dust group. On the other hand, there was no significant difference in eNOS expression between the two groups. CONCLUSIONS: We assume that the increased iNOS expression of the epithelial cells in the house dust group might result from stimulated secretion of proinflammatory cytokines during allergic responses. This further suggests profound contribution of nasal epithelial cells to modifying the airway clearance through the production of high levels of NO.     

细胞黏附分子及一氧化氮合酶在变应性鼻炎鼻黏膜中的表达

目的:对细胞黏附分子(CAM)及一氧化氮合酶(NOS)进行原位观察,探讨它们在变应性鼻炎(AR)发病中的作用。方法:采用链霉卵白素-生物素复合体(SABC)法,对AR患者及对照组手术切除的下鼻甲黏膜内细胞间黏附分子-1(ICAM-1)、血管CAM-1(VCAM-1)和淋巴细胞功能相关抗原-1(LFA-1),以及神经型NOS(nNOS)、诱导型NOS(iNOS)和内皮细胞型NOS(eNOS)进行原位检测。结果:AR下鼻甲黏膜内3种CAM表达的阳性细胞数ICAM-1为[(14.4±2.2)个/HP(×400),以下同],LFA-1为(17.2±3.3)个/HP,VCAM-1为(11.5±2.7)个/HP;对照组下鼻甲黏膜内3种CAM表达的阳性细胞数ICAM-1为(8.7±1.8)个/HP,LFA-1为(10.3±2.1)个/HP,VCAM-1为(6.9±1.8)个/HP。t值分别是11.57,10.02和8.07(均P<0.01)。AR及对照组下鼻甲黏膜内nNOS表达的阳性细胞数分别为(9.4±1.7)个/HP和(4.7±1.3)个/HP,t值为12.62,(P<0.01);iNOS表达的阳性细胞数分别为(27.5±3.2)个/HP和(4.3±1.7)个/HP,t值为36.03(P<0.01)。eNOS表达的阳性细胞数分别为(6.5±2.1)个/HP,(6.2±1.9)个/HP,t值为0.62(P>0.05)。结论:CAM在黏膜上皮、腺上皮、血管内皮以及黏膜下的各种炎性细胞等的表达,说明CAM参与AR的发生、发展。nNOS和iNOS在AR的发病过程中可能起重要作用。     

Immunolocalization of inducible nitric oxide synthase and 3-nitrotyrosine in the nasal mucosa of patients with rhinitis

Since nitric oxide (NO) can be involved in multiple physiological and pathological functions, we evaluated its possible involvement and that of peroxynitrite in the pathogenesis of rhinitis. Inferior nasal turbinates were obtained from allergic rhinitis and nonallergic rhinitis patients during corrective nasal surgery. The expressions of the inducible form of nitric oxide synthase (iNOS) and the production of peroxynitrite and its metabolite 3-nitrotyrosine were examined by immunohistochemistry in consecutive tissue sections. Each section (or tissue compartment) was given a score of 0–4 according to the labeling intensity seen, with the highest number representing the highest labeling intensity. The results showed that iNOS expression was present mainly in the mucosal epithelium, vascular endothelium, and submucosal glands. A significant difference was only observed in the labeling scores of glandular tissues of the allergic group, which had a higher iNOS labeling score. We also found that sections with a higher iNOS level did not necessarily exhibit a higher 3-nitrotyrosine labeling intensity. These data suggest that iNOS-derived NO may have a role in the pathophysiology of rhinitis, especially the glandular function of allergic nasal mucosa. Moreover, our findings suggest that the production of peroxynitrite in rhinitis patients is not dependent on the level of iNOS alone. Received: 23 September 1999 / Accepted: 4 November 1999     

沈阳市中学生变应性鼻炎的变应原分布特点

目的分析沈阳市中学生变应性鼻炎(AR)的变应原分布特点,提出对本地区青少年AR有针对性的防治策略。方法按照2006年WHO发布的《变应性鼻炎及其对哮喘的影响》(allergic rhinitis and its impacton asthma,ARIA)指南,对沈阳市2000名14～16岁中学生进行问卷调查,确定182名中学生患有AR。行皮肤点刺试验,分析其变应原,并对患者的鼻部症状行视觉模拟量表(visual analogue scale,VAS)评分和鼻结膜炎相关生活质量问卷(rhinoconjunctivitis quality of lifequestionnaire,RQLQ)进行相关性分析。结果 182例患者中,未检测出变应原者54例,检出1种变应原者15例(11.72%),检出2种变应原者19例(14.84%),检出3种及以上变应原者94例(73.44%)。屋尘螨为最常见变应原,共检出106例。鼻部症状的严重程度和生活质量呈正相关(r=0.653,P<0.001)。结论尘螨是沈阳中学生AR的主要变应原。     

我国11个城市变应性鼻炎自报患病率的相关因素分析

目的在获取我国11个中心城市变应性鼻炎自报患病率的基础上，分析社会经济和环境因素对变应性鼻炎自报患病率的影响。方法2004年9月-2005年5月，在中国2个直辖市（北京、上海）和9个省会城市（西安、沈阳、武汉、长沙、南京、杭州、广州、长春和乌鲁木齐）进行电话问卷调查，了解11个城市变应性鼻炎自报患病率。同时，获取相应城市的主要社会经济水平、气象状况和空气污染状况等指标，分析患病率与相关指标间的关系。结果①11个城市经性别和年龄校正后的变应性鼻炎自报患病率，与相应城市的主要社会经济指标（人口数量、地区生产总值、人均地区生产总值、工业生产总值和人均工业生产总值）相关性分析无统计学意义（P值均〉0．05）。②经性别和年龄矫正后的变应性鼻炎自报患病率与相应城市空气中二氧化硫浓度呈正相关关系（r2值分别为0．673和0．614，P值均〈0．05），而与其他空气污染指标的相关性无统计学意义。③经性别和年龄校正的变应性鼻炎自报患病率与相应城市的经度呈负相关关系（r^2值分别为-0．712和-0．635，P值均〈0．05），而与相应城市的纬度相关性检验，差异无统计学意义（P值均〉0．05），且与主要气象指标的相关性差异无统计学意义（P值均〉0．05）。结论11个中心城市经性别和年龄校正后的变应性鼻炎自报患病率，与空气中二氧化硫的浓度呈正相关关系，与所在城市的经度呈负相关关系，与其他社会经济和环境因素的相关关系不显著，仍需深入研究。