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。     

变应性鼻炎患者外周血白细胞及鼻黏膜诱导型一氧化氮合酶mRNA 表达的意义

目的　探讨变应性鼻炎 (allergicrhinitis,AR)患者外周血白细胞及鼻黏膜诱导型一氧化氮合酶 (induciblenitricoxidesynthase,iNOS)mRNA表达的关系。方法　选择 3 5例AR患者及 3 0例健康人的外周血白细胞。其中 8例变应性鼻炎患者鼻黏膜 ,6例正常鼻黏膜。iNOS mRNA表达采用原位杂交方法。血浆一氧化氮 (nitricoxide,NO)水平采用硝酸还原酶比色法测定。结果　健康人外周血白细胞未见iNOS mRNA表达 ,而AR患者外周血白细胞iNOS mRNA表达高度增强 ,其阳性率达 40 82 %。正常人鼻黏膜上皮、腺体及巨噬细胞可见iNOS mRNA的低度表达 ,而AR患者中上皮、腺体及巨噬细胞增生 ,并iNOS mRNA表达高度增强 (t=2 3 17,P <0 0 0 1)。AR组血浆NO水平显著高于对照组 (t=2 7 89,P <0 0 1)。结论　AR患者血浆NO水平与外周血白细胞及组织内iNOS mRNA表达高度增强有关。提示iNOS NO通路在AR的发病过程中可能起重要作用。本研究为检测体内某些信号提供了简便易行的原位杂交试验方法     

鼻息肉中一氧化氮合酶表达及其意义

目的 :了解鼻息肉 (NP)中一氧化氮合酶 (NOS)的分布特点和活性及其在NP发病中的作用。方法 :用免疫组织化学法检测 30例NP(NP组 )及 2 0例正常鼻黏膜 (正常鼻黏膜组 )中NOS的表达 ,并用分光光度计法检测其活性。结果 :NP组NOS活性为 (4.0 79± 0 .6 5 5 )U/mg蛋白 ,正常鼻黏膜组为 (1.5 2 6± 0 .310 )U/mg蛋白 ,二者间差异有统计学意义 (P <0 .0 1) ;NP组iNOS有大量细胞表达 ,分布在黏膜上皮、腺体和血管内皮细胞以及炎症细胞中 ,与正常鼻黏膜组比较 ,差异有统计学意义 (P <0 .0 1) ;而eNOS也有表达 ,但与正常鼻黏膜组比较 ,差异无统计学意义 ;NP组i NOS表达明显高于eNOS表达 ,其差异有统计学意义 (P <0 .0 1)。结论 :NP主要表达iNOS ,分布在黏膜上皮、腺体和血管内皮细胞以及炎症细胞中 ,其产生的大量一氧化氮在NP的发病过程中可能起着重要的作用     

内源性一氧化碳对变应性鼻炎豚鼠诱导型一氧化氮合酶表达的影响

目的 制备豚鼠变应性鼻炎(allergic rhinitis,AR)动物模型,研究在AR豚鼠模型中内源性一氧化碳(carbon monoxide,CO)对诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)表达的影响.方法 24只豚鼠以随机数字表法分为4组,每组6只.第1组以生理盐水处理作为正常对照组,第2(AR组)、3、4组以卵清蛋白(ovalbumin,OVA)致敏,制成AR动物模型,第3、4组再分别以血红素氧合酶1(hemeoxygenase 1,HO-1)诱导剂氯化血红素和抑制剂锌原卟啉干预处理,分别作为HO诱导组和HO抑制组,分别测定各组豚鼠血浆中碳氧血红蛋白(carboxyhemoglobin,COHb)的百分含量(用来代表血浆中CO含量),并采用实时荧光定量反转录聚合酶链反应(RT-PCR)法测定鼻黏膜中HO-1和iNOS的相对表达量.结果 第2、3、4组豚鼠AR造模成功.血浆COHb含量(x-±s,以下同)第2组(2.27%±1.13%)高于第1组(1.08%±0.24%),差异有统计学意义(q=4.10,P＜0.01);第3组(3.17%±0.68%)高于第2组,差异有统计学意义(q=3.12,P＜0.05).鼻黏膜中HO-1、iNOS的相对表达量(x-±s,以下同)第2组[分别为(7.80±1.60)×10~(-3)和(5.81±0.05)×10~(-3)]高于第1组[分别为(1.96±0.71)×10~(-3)和(0.97±0.05)×10~(-3)],差异有统计学意义(q值分别为5.52、7.21,P值均＜0.01),第3组[分别为(11.89±4.78)×10~(-3)和(7.42±0.70)×10~(-3)]高于第2组,差异有统计学意义(q值分别为3.86、2.22,P值均＜0.05),第4组[分别为(3.82±0.98)×10~(-3)和(2.34±0.04)×10~(-3)]低于第2组,差异有统计学意义(q值分别为3.76、5.18,P值均＜0.05).结论 内源性CO在AR中影响iNOS的表达.     

Toll样受体-4介导诱发型一氧化氮合酶mRNA在鼻黏膜上皮细胞中的表达

目的 :探讨Toll样受体 4 (TLR 4 )对诱发型一氧化氮合酶 (iNOS )在鼻黏膜上皮细胞中表达的上调作用。方法 :应用核酸分子原位杂交技术检测 30例慢性鼻窦炎患者 (鼻窦炎组 )和 2 1例健康者 (对照组 )鼻黏膜上皮细胞中TLR 4和iNOSmRNA的表达。结果 :TLR 4和iNOSmRNA在鼻窦炎组鼻黏膜上皮细胞中的表达 ,分别为 (0 .14 33± 0 .0 184 0 ) /A和 (0 .136 8± 0 .0 0 76 5 ) /A ,分别与对照组 [(0 .10 2 4± 0 .0 1133) /A和 (0 .0 72 4± 0 .0 136 4 ) /A ]比较 ,差异有统计学意义 (均P <0 .0 1) ,且二者表达呈正相关 (r =0 .4 35 ,P <0 .0 1)。结论 :鼻黏膜上皮细胞通过TLR 4识别病原微生物 ,上调iNOSmRNA的表达 ,增加NO的合成 ,以杀伤、清除病原菌 ,增强宿主防御和免疫应答。     

缺氧对鼻粘膜上皮细胞中诱发型一氧化氮合酶的影响及其意义

目的 :探讨鼻粘膜上皮细胞应答局部组织缺氧 ,合成分泌诱发型一氧化氮合酶 (iNOS)的变化 ,以及地塞米松对此的影响及意义。方法 :将人鼻粘膜上皮细胞在常氧和缺氧状态下进行无血清原代细胞培养 ,并加入不同浓度的地塞米松共同孵育 ,采用流式细胞仪观察常氧和缺氧条件下上皮细胞动力周期的变化 ,采用原位杂交的方法检测iNOSmRNA的变化。结果 :缺氧条件下上皮细胞的动力周期延长 ;在缺氧 3h后iNOSmRNA水平开始升高 ,12h达高峰 ,2 4h后下降 ,4 8h几乎消失 ;加入地塞米松后 ,降低这种升高的水平。但 4 μg L以上的浓度 ,并不进一步降低被缺氧升高的iNOSmRNA水平。结论 :缺氧诱发鼻粘膜上皮细胞合成高水平的iNOSmR NA ,应用一定浓度的地塞米松能降低这种作用     

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     

鼻息肉鼻内镜术后诱导型一氧化氮合酶的检测

目的　检测诱导型一氧化氮合酶 (iNOS)在鼻息肉 (NP)鼻内镜术中术后的表达情况 ,探讨iNOS在预测NP的复发及客观判断内镜手术疗效中的作用。方法　取 5 0例NP鼻内镜术中术后NP组织、中鼻道黏膜组织和 37例行下鼻甲切除的下鼻甲黏膜组织 (对照组 ) ,应用免疫组化染色方法 ,分别检测两组组织中iNOS的表达情况。结果　①iNOS在 5 0例鼻内镜术中NP组织的上皮及腺上皮细胞中均有表达 ,在炎症细胞也有表达 ;在 37例对照组鼻黏膜上皮及腺上皮组织均无表达 (P <0 .0 1)。②NP组在术后 1～ 4个月中鼻道黏膜iNOS表达机率逐渐减少 ,大多数可见弱阳性表达。结论　iNOS在NP发生发展中发挥着重要作用 ;NP鼻内镜术后的iNOS动态监测可以作为制定手术疗效客观标准的参考和预测NP复发的高危因素之一     

鼻黏膜中一氧化氮合酶表达及其意义

目的了解正常鼻黏膜中一氧化氮合酶的分布特点及活性,并分析其在鼻黏膜中的生理作用。方法应用免疫组化法对20例正常鼻黏膜中内皮型一氧化氮合酶和诱导型一氧化氮合酶的表达情况进行检测,并应用分光光度法检测一氧化氮合酶活性,然后分析其表达特征,探讨其生理意义。结果正常鼻黏膜组织中有内皮型一氧化氮合酶表达,分布于黏膜上皮、腺体和血管内皮细胞;诱导型一氧化氮合酶偶见细胞表达;两者表达水平差异有显著性意义(P<0.01)。一氧化氮合酶活性1.526±0.310U/mgPro。结论鼻黏膜中主要表达内皮型一氧化氮合酶,分布于黏膜上皮、腺体和血管内皮细胞。由该酶产生的一氧化氮在鼻腔的正常生理功能中可能发挥重要作用。     

慢性鼻窦炎鼻息肉伴发嗅觉障碍病人iNOS在嗅粘膜中的表达

目的 探讨诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)在慢性鼻窦炎鼻息肉嗅觉障碍发病过程中的作用。方法 30例慢性鼻窦炎嗅觉障碍病人经临床分型分期,行嗅觉功能检查后,取嗅粘膜标本,行iNOS的免疫组化染色,光镜观察。结果 慢性鼻窦炎鼻息肉2型2,3期和3型的嗅粘膜中iNOS的表达显著高于1型和2型1期(P<0.05)。并与嗅觉障碍的严重程度呈正相关。结论 iNOS可能参与了慢性鼻窦炎所致嗅觉障碍的嗅觉损害过程,损害程度与慢性耳窦炎鼻息肉的临床分型分期有关。     

诱生型一氧化氮合酶在慢性上颌窦炎黏膜的表达

目的 观察诱生型一氧化氮合酶(inducible nitric oxide synthase，iNOS)在慢性鼻窦炎病人鼻窦粘膜中的表达。方法 将实验分为两组：对照组为正常成人，无鼻窦病变。实验组为确诊为慢性上颌窦炎的病人。在局麻鼻内镜下行鼻窦黏膜活检，石蜡包埋、切片，用免疫组织化学的方法观察iNOS在两组上颌窦黏膜中的表达。结果 iNOS在正常上颌窦炎黏膜呈阴性表达，在慢性上颌窦炎黏膜呈阳性表达，阳性区域主要在黏膜上皮细胞。结论 iNOS在慢性上颌窦炎黏膜呈阳性表达，提示一氧化氮在慢性上颌窦炎的发病机制中起重要作用。     

豚鼠变应性鼻炎模型鼻黏膜的病理改变

目的：观察变应性鼻炎鼻黏膜的病理变化。方法：用橄榄油将甲苯-2,4-二异氰酸酯配成浓度为10%的溶液作为致敏剂,滴鼻,建立豚鼠变应性鼻炎动物模型8例。8例正常豚鼠作对照。致敏结束、模型成功后,取两组鼻黏膜组织,光镜下观察病理变化。结果：模型组鼻黏膜上皮脱落,杯状细胞增生,鳞状上皮组织转化,上皮坏死,固有层和黏膜下层腺体增生,血管扩张,组织水肿,并见特征性的嗜酸性粒细胞、肥大细胞浸润,这种病理改变与变应性鼻炎的临床表现大致吻合。对照组鼻黏膜上皮层为假复层纤毛柱状上皮,连续、完整、清晰,可见正常的黏膜上皮层、固有层和黏膜下层。结论：变应性鼻炎鼻黏膜出现特征性的炎症病理改变。     

Lipopolysaccharide-induced expression of inducible nitric oxide synthase in the guinea pig organ of Corti

The purpose of the investigation was to ascertain whether inoculation of bacterial lipopolysaccharide (LPS) into the cochlea of the guinea pig could elicit formation of inducible nitric oxide synthase (iNOS). Immunohistochemical study revealed that immunoreactivity to iNOS was seen below outer hair cells representing nerve fibers and synaptic nerve endings. iNOS-staining could also be observed in phalangeal dendrites of Deiter’s cells pointing to the cuticular membrane, Hensen’s cells and on stria vascularis 48 h after inoculation with LPS. Immunohistochemical investigation with a specific anti-nitrotyrosine antibody also revealed intense immunoreactivity identical to that of iNOS, suggesting formation of peroxynitrite in the organ of Corti by the reaction of NO with O₂⁻. On the basis of these findings, it can be concluded that NO together with O₂⁻, which form the more reactive peroxynitrite, are the most important pathogenic agents in LPS-induced damage of cochlea in the guinea pig.     

Distribution of nitric oxide in the nasal mucosa of the rat: A histochemical study

We evaluated the distribution of nitric oxide (NO) in the rat nasal mucosa using nicotineamide adenosine dinucleotide phosphate (NADPH)-diaphorase histochemistry. The NADPH-diaphorase positive nerve fibers in the nasal mucosa were observed around blood vessels and submucosal glands and in sphenopalatine ganglions. Strong positive reactions for NADPH-diaphorase were observed in ganglions as compared with the other tissues. In septal and turbinate mucosa, positive reactions for NADPH-diaphorase were mainly seen in the anterior portion, and a few positive reactions were observed in the posterior portion. No positive reactions for NADPH-diaphorase were demonstrated in the sinus mucosa. These results suggest that NO may be related to regulation of blood flow, glandular secretion and neurotransmission, and also that NO may play an important role in the defence mechanism of the upper airway system against external environments.     

Reduction of neuronal and inducible nitric oxide synthase gene expression in patients with cystic fibrosis

As a consequence of diminished nitric oxide synthase (NOS) protein concentration, the airway concentration of nitric oxide (NO) is reduced in patients with cystic fibrosis (CF). This appears to lead to a reduced elimination of such microorganisms as Pseudomonas aeruginosa. The objective of this study was to analyze whether inducible (iNOS), endothelial (eNOS) and neuronal (bNOS) NOS are reduced at mRNA level and if so whether this is caused directly by the defective CF transmembrane conductance regulator (CFTR). Nasal polyps from three patients with CF and four otherwise healthy patients were obtained. The expression of the three NOS isoenzymes was quantified using real-time PCR. The iNOS expression was assessed in colon carcinoma cells (CaCo) transfected with a normal and a mutated (DeltaF508) CFTR. In CF patients, iNOS mRNA expression was 10- to 20-fold and bNOS gene expression was one-fifth to one-tenth that in control patients (P < 0.001). In CaCo cells, iNOS gene expression under basal and endotoxin-stimulated conditions did not differ between cells transfected with a mutated CFTR and those transfected with an intact CFTR. This observation suggests that cystic fibrosis is associated with reduced iNOS and bNOS gene expression in nasopharyngeal tissue, possibly disturbing the barrier against infective agents already at the site of entrance. Received: 7 March 2001 / Accepted: 26 September 2001     

Regulation of mucociliary motility by nitric oxide and expression of nitric oxide synthase in the human sinus epithelial cells

OBJECTIVES: This study was performed to investigate the changes in ciliary beat frequency (CBF) after treatment with Larginine in the human sinus mucosa and to determine the distribution of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in the healthy sinus mucosa. STUDY DESIGN/METHODS: CBF was measured in the sphenoid sinus mucosa of 12 patients who underwent trans-septal trans-sphenoidal hypophysectomy for the treatment of pituitary gland tumor. CBF was measured over 24 hours in Dulbecco's modified Eagle's medium (DMEM) after treatment with L-arginine, its inactive spatial isomer D-arginine, or an NOS inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME). DMEM without treatment with these materials was used as a control. Other pieces of the mucosa were exposed to L-NAME and its inactive spatial isomer D-NAME after preincubation with L-arginine. The specimens were immunohistochemically stained for iNOS and eNOS. RESULTS: CBF increased 24 hours after treatment with L-arginine as compared with control groups. CBF increased in proportion to the increasing concentrations of L-arginine. There was no significant change after treatment with D-arginine or L-NAME. CBF increased after treatment with L-arginine at 30 minutes and maintained for 24 hours. L-NAME inhibited the increase in CBF by L-arginine, but D-NAME showed no such effect. Immunoreactivity to both iNOS and eNOS was frequently observed in the ciliated epithelial cells and was stronger to eNOS than to iNOS. CONCLUSIONS: From the results of this study it is suggested that nitric oxide (NO) produced by iNOS and eNOS using L-arginine may increase CBF in the healthy sinus mucosa and that NO may have a regulatory function in ciliary motility in the human sinus mucosa.     

诱导型一氧化氮合酶在变应性真菌性鼻-鼻窦炎黏膜中的表达

目的 :通过对变应性真菌性鼻 鼻窦炎窦腔病变黏膜 (AFRS)和慢性鼻窦炎 (CRS)病变黏膜中诱导型一氧化氮合酶 (iNOS)的定位和半定量研究 ,探讨AFRS和CRS发病机制的差异 ,一氧化氮 (NO)的生成情况和意义。方法 :用免疫组化 (S P法 )的方法对 2 8例AFRS和 6例CRS标本中iNOS进行定位和半定量检测。并且用Ridit分析进行统计分析。结果 :iNOS广泛存在于上皮组织、血管内皮、平滑肌细胞、黏膜下浆液性腺体和炎症细胞中 ,以细颗粒或粗颗粒存在于细胞质中 ,细胞膜上偶见。细胞核及间质细胞内无iNOS表达。AFRS黏膜与CRS黏膜之间的表达差异有显著性意义 (P <0 .0 1)。结论 :iNOS在AFRS窦腔黏膜中呈大量表达 ,iNOS对AFRS发病起重要作用 ;AFRS与CRS可能是不同的两个疾病     

Endothelial nitric oxide synthase/soluble guanylate cyclase system in human nasal polyps

The aim of our study was to analyze the level of expression of the endothelial nitric oxide synthase (eNOS)/soluble guanylate cyclase (sGC) system in nasal polyps and control nasal mucosae. The study was performed in polyps from 15 patients and nasal mucosae from 11 subjects operated on the nasal septum (control group). The expression of endothelial nitric oxide synthase (eNOS) and soluble guanylate cyclase (sGC) was determined in nasal mucosae. Western blot analysis demonstrated that eNOS protein was overexpressed in the nasal polyps with respect to control nasal mucosae. Immunohistochemistry also demonstrated that the vascular endothelium of nasal polyps contained higher amounts of eNOS protein than control nasal mucosae. Moreover, the ₁ subunit of sGC was also overexpressed in the nasal polyps, which was associated with an increased content of cyclic GMP in the nasal polyps with respect to nasal control mucosae. In human nasal polyposis, there is an overexpression of the eNOS/sGC system. Further studies are needed to assess whether this overexpression is involved in the genesis of nasal polyposis.     

白噪声对豚鼠耳蜗核一氧化氮合酶活性的影响

采用硫辛酰胺脱氢酶组织化学方法及图象分析技术，研究白噪声暴露后豚鼠耳蜗核一氧化氮合酶（ＮＯＳ）神经元及ＮＯＳ活性的变化与听阈的关系，探讨豚鼠耳蜗核ＮＯＳ神经元在白噪声损伤过程中可能的作用。结果表明，白噪声暴露后耳蜗核ＮＯＳ阳性神经元的数量及染色强度明显增加，２周达到高峰，３￣４周持续高表达，至５周有所恢复，仍高于正常水平。白噪声暴露后７ｄ以内，耳蜗核ＮＯＳ活性与ＡＢＲ阈值有分离现象，７ｄ后，ＮＯＳ