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     

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。     

慢性鼻炎鼻黏膜中一氧化氮合酶的表达

目的比较正常鼻黏膜和慢性鼻炎鼻黏膜中一氧化氮合酶(NOS)的分布,探讨NOS与慢性鼻炎的关系.方法应用还原型尼克酰胺嘌呤二核苷酸磷酸-黄递酶(NADPH-d)组织化学染色与免疫组织化学技术,测定内皮型NOS(eNOS)和诱导型NOS (iNOS) 在正常组和慢性鼻炎组鼻黏膜中的分布和表达.结果eNOS免疫组化显示正常鼻黏膜和慢性鼻炎鼻黏膜NOS均呈阳性反应,主要分布于表层上皮细胞,血管内皮细胞胞浆;iNOS免疫组化则显示慢性鼻炎表层上皮细胞,血管内皮细胞胞浆NOS阳性,部分炎性细胞亦呈阳性反应;而正常鼻黏膜NOS呈阴性.酶组织化学染色显示,NOS有同样的分布部位.结论正常鼻黏膜存在NOS分布,慢性鼻炎鼻黏膜中iNOS活性明显高于正常,由iNOS产生的NO和慢性鼻炎的鼻分泌物增多和黏膜水肿有关.     

一氧化氮及其合酶在鼻息肉中的表达

目的：探讨一氧化氮(NO)及其合酶(NOS)在鼻息肉中的表达，以及NO在鼻息肉发病中的作用。方法：用免疫组织化学及原位杂交方法研究诱导型一氧化氮合酶(iNOS)及内皮型一氧化氮合酶(eNOS)在鼻息肉中的表达，同时用原位杂交方法研究iNOS mRNA的表达，并用硝酸还原酶法研究NO在鼻息肉中的产生情况。结果：鼻息肉组织中eNOS主要分布于上皮、腺体细胞及血管内皮细胞，其染色强度稍强于对照组。iNOS在上皮细胞呈现较强的阳性染色，在息肉组织内主要表达于散在的炎症细胞。结论：eNOS活性增高可能与鼻息肉发病中血管过度扩张、腺体病理性分泌增多等有关。iNOS生成的较高浓度的NO在鼻息肉的病理过程中可能起到促进炎症发展的作用。     

鼻黏膜诱发型一氧化氮合酶的表达与鼻内镜术后鼻黏膜转归关系的探讨

目的:观察慢性鼻窦炎(CRS)患者鼻黏膜中诱发型一氧化氮合酶(iNOS)的表达水平,探讨iNOS在CRS炎症中的作用以及与鼻内镜术后鼻黏膜转归的相关性.方法:根据血清总免疫球蛋白E(TIgE)水平将所有CRS病例分为TIgE水平高的A组(TIgE≥90 kU/L)和TIgE不高的B组(TIgE＜90 kU/L),2组皆于鼻内镜术(ESS)前取病变鼻黏膜检测iNOS的表达并参照Lund-Kennedy系统对鼻窦冠状位CT进行评分.追踪2组病例术后同侧术腔完成上皮化的时间.分析CT评分以及鼻黏膜iNOS表达与术腔上皮化时间的相关性.结果:A组的上皮化时间与鼻黏膜iNOS的表达之间呈正相关(P＜0.05);与鼻窦CT评分无明显相关.B组的上皮化时间与鼻窦CT评分呈正相关(P＜0.05),而与鼻黏膜iNOS的表达无相关.结论:iNOS对预测CRS术后术腔恢复有价值,TIgE高的情况下,iNOS的表达强度与术后完成上皮化的时间呈正相关.伴变应性因素的CRS患者,术前鼻黏膜高强度的iNOS表达,很可能预示着术后上皮化需要较长的过程.     

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

OBJECTIVE: To investigate the relationship between expression of leukocyte inducible nitric oxide synthase (iNOS)-mRNA and allergic rhinitis (AR). METHOD: Thirty-five patients with AR and 30 healthy controls were included in this study. Expression of iNOS-mRNA in peripheral blood leukocyte was detected by in situ hybridization. NO in plasm was measured by nitrate reductase. Expression of iNOS-mRNA in nasal mucosal was detected in 8 patients with AR and 6 healthy controls. RESULT: No expression of leukocyte iNOS-mRNA in healthy controls was found. In AR patients, the positive cells were significantly increased, the positive rate reached 40.82%. Expression of iNOS-mRNA was localized at the epithelium, gland and macrophage in healthy controls. Hyperplasia and expression of iNOS-mRNA increased at epithelium, gland and macrophage in the AR patients(t = 23.17, P < 0.001). The level of plasm NO in AR group was higher than that in healthy control group (t = 27.89, P < 0.001). CONCLUSION: There is a relationship between expression of leukocyte iNOS-mRNA and the level of plasm NO in AR patients. The study provides an easy method of in situ hybridization for detecting some signal in body.     

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     

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.     

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.     

低氧刺激鼻息肉黏膜上皮细胞炎性因子变化的初探

目的:研究低氧刺激慢性鼻窦炎伴鼻息肉（CRSwNP）与正常鼻黏膜上皮细胞炎性因子的变化与异同,探讨其在CRSwNP发病机制中的作用。方法:选择2015年6月至2018年1月在吉林大学中日联谊医院就诊的68例CRSwNP患者,其中男性36例,女性32例,年龄（45.2±12.5）岁（ xˉ±s,下同）,患者的...     

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     

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