一氧化氮在鼻息肉及鼻黏膜慢性炎症中的检测

目的检测正常鼻黏膜一氧化氮(nitric oxide, NO)分布特点以及鼻息肉患者息肉组织中NO含量,探讨其在鼻粘膜炎症过程中的意义。方法采用重氮化反应法(Greiss反应法)检测20例鼻息肉患者(A组)的息肉组织中和下鼻甲黏膜中NO的表达,并与12例健康人(B组)下鼻甲、钩突及嗅裂粘膜对照:A、B两组手术前静脉血中NO含量作比较。结果A组患者血清、息肉组织及下鼻甲黏膜中NO含量分别为(63．8±18．7)μmol／L、(105．6±23．5)μmol／L、(81．4±20．4)μmol／L;B组血清、下鼻甲、钩突及嗅裂黏膜中NO含量分别为(63．3±23．0)μmol／L、(54．5±18．1)μmol／L、(83．0±23．7)μmol／L、(62．3±24．4)μmol／L。两组血清中NO含量差别无统计学意义(P<0．05);鼻息肉组织中NO含量高于鼻黏膜以及正常鼻黏膜组织NO含量(P<0．05);鼻息肉患者下鼻甲黏膜中NO含量高于正常鼻黏膜(除钩突黏膜外)NO含量(P<0．05),正常鼻黏膜中NO含量分布有差别(P<0．05)。结论NO是一种具有广泛生物活性的物质,在鼻息肉发病机理中和导致鼻黏膜持续性炎症中有重要作用。     

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。     

诱导型一氧化氮合酶mRNA在分泌性中耳炎中表达的意义

目的探讨分泌性中耳炎(secretory ototis media，SOM)患者中耳积液及外周血白细胞诱导型一氧化氮合酶(inducible nitric oxide synthase，iNOS)mRNA表达及在SOM发病过程中的意义，以及与细菌感染和免疫介导的关系。方法随机取45例SOM患者及30例健康人的外周血白细胞，并抽取SOM患者患耳中耳积液，用原位杂交方法检测iNOS—mRNA。结果健康人外周血白细胞未见iNOS—mRNA表达。SOM患者外周血白细胞iNOS—mRNA表达：急性组阳性率为63．64％，镜下阳性细胞率为60．3％；亚急性组阳性率为8．7％，镜下阳性细胞率为72．5％；SOM患者中耳积液iNOS—mRNA表达：急性组阳性率为45．45％，镜下阳性细胞率为80％；亚急性组阳性率为52．17％，镜下阳性细胞率为84％。SOM患者外周血白细胞及中耳积液中iNOS—mRNA表达高度增强，其中在急性组外周血白细胞中表达显著高于亚急性组，而中耳积液iNOS—mRNA表达阳性率及阳性细胞率在急性组与亚急性组中表达强度无显著性差异。结论诱导型一氧化氮合酶(iNOS)及诱导型一氧化氮合酶——氧化氮(iNOS—NO)通路在SOM的发病、中耳积液的形成过程中可能起着重要作用。     

变应性鼻炎患者血清氧化应激状态研究

目的 检测变应性鼻炎(allergic rhinitis,AR)患者外周血血清中氧化应激标志物,探讨氧化应激反应在AR发病机制中的作用.方法 留取23例健康人、48例AR患者外周血血清,分别检测氧化应激标志物一氧化氮(nitric oxide,NO)、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、总一氧化氮合酶(total nitric oxide synthase,TNOS),脂质过氧化产物丙二醛(malondidehyde,MDA),以及体内主要抗氧化酶超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)水平.采用SPSS 13.0软件对数据进行分析.结果 AR患者血清NO水平((x)±s,以下同)为[(97.92±73.42)μmol/L]较健康对照组[(64.04±29.54)μmol/L]明显升高(t=-0.281,P<0.05);iNOS与TNOS比值(0.51±0.11)较健康对照组(0.45±0.15)明显升高(t=-2.061,P<0.05);血清GSH-Px活性[(258.24±45.25)U/(ml·min)]较健康对照组[(215.11±47.62)U/(ml·min)]明显升高(t=-2.2349,P<0.05)).而AR患者血清SOD活性、MDA含量较健康对照组分别有升高和降低趋势,但差异无统计学意义(Z=-1.656,t=1.922,P值均>0.05).结论 氧化应激反应参与AR病理生理过程,而iNOS-NO通路在AR的发病过程中可能发挥着更为重要的作用.     

分泌性中耳炎中耳积液中γ干扰素测定的临床意义

目的:探讨分泌性中耳炎(SOM)患者的中耳积液中IFNγ浓度测定的临床意义。方法:用双抗夹心酶联免疫吸附法检测51例(56耳)SOM患者(中耳炎组)的中耳积液、血清及24例正常成人(对照组)血清中IFNγ的浓度。结果:中耳炎组积液中IFNγ的浓度明显高于其血清中的浓度(P<0.01);中耳炎组与对照组血清中的浓度差异无统计学意义(P>0.05);SOM慢性期IFNγ浓度明显高于急性期(P<0.05);首次穿刺及第2次穿刺积液中IFNγ的浓度差异无统计学意义(P>0.05);而第3次或3次以上穿刺积液中IFNγ的浓度则明显升高(P<0.01)。结论:中耳积液中的IFNγ可由中耳腔局部产生,而非单纯由血液中渗透而来;中耳积液中IFNγ的高浓度可作为SOM转为慢性病程或迁延不愈的参考。     

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     

转化生长因子β在分泌性中耳炎患者中耳积液和血清中的表达及意义

分泌性中耳炎(SOM)的发病机制还不十分清楚.一般认为咽鼓管阻塞、中耳负压导致血管内液体渗漏形成中耳积液.后来有学者发现咽鼓管阻塞并非是引起SOM的必要条件,而中耳积液中炎症递质和炎性细胞的发现,证实SOM是一炎症反应和渗出的结果.     

纤维蛋白原含量与分泌性中耳炎病情迁延的关系

目的:探讨中耳积液中纤维蛋白原含量与分泌性中耳炎病情迁延的关系。方法:用凝固法对98例成人患者中耳积液中纤维蛋白原进行动态检测。结果:98例患者经鼓膜穿刺治疗后62例未愈,治愈组和迁延组纤维蛋白原检测阳性率分别为30.56%和72.58%,P<0.01;浓度分别为(0.350±0.124)g/L和(0.568±0.206)g/L,P<0.05。迁延组中52例患者第2次穿刺治疗中耳积液中纤维蛋白原浓度为(1.241±0.146)g/L,明显高于第1次[(0.685±0.251)g/L],P<0.01。结论:纤维蛋白原含量与分泌性中耳炎病情迁延密切相关,可能在粘连性中耳炎形成中起重要作用。     

细胞因子及白细胞介素-1β与内毒素在分泌性中耳炎中耳积液中的表达及意义

目的 :探讨分泌性中耳炎 (SOM )中耳积液中内毒素 (ET)、白细胞介素 1β(IL 1β)、正常T细胞表达和分泌、活化时调节的趋化因子 (RANTES)的表达以及它们在SOM发病中的作用。方法 :对 5 3例 72耳SOM患者行鼓膜穿刺 ,获得的中耳积液标本行细菌培养 ,然后采用鲎试验动态浊度法、放射免疫法以及双夹心抗体酶联免疫吸附法检测中耳积液中ET、IL 1β和RANTES的浓度。 结果 :①中耳积液中ET、IL 1β和RANTES阳性率分别为 80 .9%、77.8%和 70 .8% ,平均浓度为 (35 .2± 5 1.6 )EU/ml,(1.10± 0 .84 ) μg/L ,(0 .5 2± 0 .4 3) μg/L。②三者在黏液性积液中的含量高于浆液组 (P <0 .0 5 ) ;病程长者 ,ET、RANTES浓度也较高 (P <0 .0 5 ) ;细菌培养阳性中耳积液中三者的浓度亦明显高于细菌培养为阴性的积液 (P <0 .0 1)。③积液中ET含量与IL 1β呈显著正相关性 (r =0 .74 ,P <0 .0 1) ,IL 1β与RANTES之间也呈显著正相关 (r =0 .4 8,P <0 .0 1)。结论 :ET、IL 1β与RANTES参与了SOM发病的免疫机制 ,与鼓室内炎症反应的迁延 ,促使积液类型转化有关     

细胞因子、IgE及一氧化氮在小儿分泌性中耳炎中耳积液的表达

目的 :探讨细胞因子、IgE及一氧化氮 (NO)在小儿分泌性中耳炎的发生及转归中的作用。 方法 :检测 70例 (1 2 9耳 )分泌性中耳炎患儿 (患儿组 )血浆及中耳积液中细胞因子、IgE及NO含量 ,并以 30例健康儿童作对照。结果 :患儿组白细胞介素 (IL 2、4、6、8、1 0 )、肿瘤坏死因子 α(TNF α)、IgE及NO在中耳积液中含量较血浆中高 (P <0 .0 1 ) ;患儿组的血浆含量较对照组高 (P <0 .0 5 )。病程短者中耳积液中IL 2、IL 4的含量较病程长者高 ;而病程长者IL 6、IL 8、IL 1 0、TNF α、IgE及NO的表达较病程短者增加 (均P <0 .0 5 )。结论 :在儿童分泌性中耳炎的血浆及中耳积液中细胞因子、IgE及NO表达增强 ,细胞因子参与介导中耳局部的炎性反应 ,调节局部的免疫反应 ,在分泌性中耳炎的发生及转归中产生重要作用     

Glucocorticoids reduce nitric oxide concentration in middle ear effusion from lipopolysaccharide induced otitis media

Objective

Otitis media with effusion (OME) is a common childhood disease that is characterized by an accumulation of fluid in the middle ear. Chronic OME can also lead to sensorineural hearing loss (SNHL). Nitric oxide (NO), an inflammatory mediator (IM) of OME, is a free radical known to regulate cell proliferation, cell death, and angiogenesis. Previous studies have shown that nitric oxide may cause SNHL through outer hair cell (OHC) cytotoxicity. This experiment was designed to determine whether glucocorticoids, dexamethasone, fluticasone propionate, or rimexolone, can reduce the concentration of NO in middle ear effusion (MEE).

Methods

Fifty-three chinchillas were divided into 7 groups, vehicle vs. each glucocorticoid at 0.1% and 1.0% concentrations. Due to anesthesia complications, N ranged from 6 to 9 per group. Two hundred microlitres of each test article was injected into the bullae of each animal. Two hours later, lipopolysaccharide (LPS) (0.3 mg in solution) was added. Test articles were re-administered at 24 and 48 h post-LPS induction. After 96 h, animals were euthanized and the MEE was collected.

Results

All three glucocorticoids numerically reduced NO concentration in the middle ear when administered at 0.1%, but only FP showed a significant reduction. At 1.0% concentrations, all 3 steroids significantly reduced NO concentration.

Conclusion

This study suggests that glucocorticoid treatment reduces NO concentration in the MEE and may protect the ear from the SNHL caused by NO.     

Nitric oxide contributes to control of effusion in experimental otitis media

OBJECTIVES/HYPOTHESIS: Nitric oxide (NO) is a small, short-lived free radical involved in cellular signaling and known to play a role in inflammation. It is generated on demand by the enzyme nitric oxide synthase (NOS) on arginine. We have previously found that mRNA encoding NOS is produced in the middle ear during otitis media. The role of NO was therefore explored in an experimental model of immune-mediated otitis media. STUDY DESIGN AND METHODS: Guinea pigs were systemically immunized and later challenged in the middle ear with the same antigen. One ear of each animal was challenged with antigen alone. In the opposite ear, antigen was combined with a potent inhibitor of NOS, N(G)-amino-L-arginine (L-NAA). After survival for 24, 48, or 72 hours, the middle ears were evaluated for otitis media. RESULTS: Inhibition of NOS resulted in significantly increased middle ear effusion at all three time periods. This increase was blocked by the addition of excess 1-arginine, which bypasses the inhibitory effects of L-NAA. The infiltration of cells into the middle ear lumen and the hyperplasia of the middle ear mucosa were unaffected by L-NAA administration. CONCLUSIONS: The results suggest that NO is involved in regulating the permeability of the middle ear vascular, the transudation of serum into the middle ear mucosa, and/or the movement of extracellular fluid across the middle ear mucosal epithelium.     

Concentration of nitric oxide metabolites in middle ear effusion.

Free radicals such as nitric oxide (NO) seem to be important in the pathogenesis of otitis media with effusion (OME). NO can be quantitated by measuring its metabolites, nitrate (NO(3)(-)) and nitrite (NO(2)(-)). The purpose of this study is to determine the concentrations of NO in human middle ear effusion (MEE). Samples of human MEE were collected at the time of myringotomy and tympanostomy tube insertions. The type of MEE was classified as serous (SOM), mucoid (MOM) or purulent (POM) at the time of surgery. Samples of MEE were assayed for NO metabolites (nitrate and nitrite) with colorimetric assay (Griess method). Concentrations of NO metabolites were highest in MOM followed by SOM and POM. This study suggests that NO is present in human MEE and may play an important role in the pathogenesis of OME.     

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。     

Heme oxygenase and nitric oxide synthase in human middle ear epithelium indicates local carbon monoxide and nitric oxide production

The gas mixture of the middle ear differs from that of the atmosphere, a fact that has been attributed to gas exchange across the middle ear mucosa. Several diseases of the middle ear seem to be related to impaired ventilation together with conjunctional changes in pressure and gas composition. Carbon monoxide (CO) and nitric oxide (NO) have recently been shown to be endogenously produced in the human lung as well as in the nasal airways. The production of CO and NO is enzymatically regulated by heme oxygenase (HO) and NO synthase (NOS), respectively. These enzymes display isoforms that are both constitutively expressed [HO-2, endothelial NOS (eNOS), neuronal NOS (nNOS)] and inducible [HO-1, inducible NOS (iNOS)] following different types of stimulation. The present study was designed to investigate the presence of HO-1, HO-2 and eNOS in the middle ear epithelium, using immunocytochemistry. Specimens from human middle ear mucosa obtained at autopsy and during surgery revealed HO-1-, HO-2- and eNOS-like immunoreactivity, indicating the possibility of local CO and NO production in the middle ear. If this assumption is true, it may affect our understanding of middle ear physiology and give new insights into the mechanisms behind middle ear pathology.     

一氧化氮及两种细胞因子在分泌性中耳炎中耳积液中的表 …

目的 探讨一氧化氮（ＮＯ）和白细胞介素－８（ｉｎｔｅｒｌｅｕｋｉｎ－８,ＩＬ－８）肿瘤坏死因子－α（ｔｕｍｏｒｎｅｃｒｏｓｉｓｆａｃｔｏｒ－α,ＴＮＦ－α）在分泌性中耳炎发生和转归中的作用。方法 检测５８例（８０耳）分泌性中耳炎患者血浆和中耳积液中ＮＯ,ＩＬ－８及ＴＮＦ－α,并用２３例健康人血浆作对照,结果 ＮＯ,ＩＬ－８及ＴＮＦ－α在中耳积液中阳性表达率分别为１００％,８２．２％和７１．４％,患     

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     

Involvement of nitric oxide synthase in the physiology and pathophysiology of facial nerve function and dysfunction

To date few reports have discussed the presence and function of nitric oxide (NO) in structures of the facial nerve. We performed nicotinamide adenine dinucleotide phosphate (NADPH-d)-diaphorase-histochemistry and immunohistochemistry on the intratemporal portion of the facial nerve, including the geniculate ganglion, of guinea pigs using specific antibodies to the three known isoforms of NO synthase and soluble guanylyl-cyclase (sGC). Normal facial nerves were compared to those treated intratympanically with bacterial lipopolysaccharides (LPS) and tumor necrosis factor-α (TNF-α). Both constitutive NOS isoforms and sGC could be detected in the bipolar ganglion cells of normal animals, while the inducible isoform (iNOS or NOS II) was not found. Endothelial NOS (NOS III) and sGC were present in blood vessels and were predominantly found in the perineurial sheath and less in the endoneurium. sGC could be detected in all fibers in a cross section of the facial nerve. LPS and TNF treatment led to the detection of iNOS in the perikaryia of the geniculate ganglion and the perineural sheath. These findings imply that NO may be involved in neurotransmission at least in the visceroafferent system. NO regulates vascular tone of nutrient blood vessels in the perineural sheath and endoneurium. The presence of sGC indicates that NO acts via its second messenger cGMP. NOS II expression may be a contributing factor to facial nerve palsy via two different mechanisms: NOS II-generated NO may lead to an overstimulation of the visceroefferent nerve fibers and motor fibers of the facial nerve. Dysregulation in facial nerve blood vessels could lead to edema and elevated pressure on the nerve within its osseous canal. Received: 13 April 1999 / Accepted: 12 August 1999