肝硬化大鼠脊髓内一氧化氮合酶阳性神经元分布的研究

目的探讨肝硬化对神经系统所产生的影响及肝硬化大鼠在脊髓中NOS量效关系,即肝硬化大鼠脊髓内一氧化氮合酶分布的研究。方法采用NADPH鄄d组化法及LeicaQ500IW图像分析系统对肝硬化大鼠脊髓颈、胸、腰段一氧化氮合酶阳性细胞进行数量及灰度的测定。结果正常组与肝硬化组分为两个等级:低密区和高密区,低密区灰度定为60,高密区为90,两组Wistar大鼠NOS阳性神经元灰度均为60,差异无显著性。在脊髓灰质区,一氧化氮合酶阳性细胞主要分布在中央管周围即脊髓板层第Ⅹ层和中间外侧核。NADPH鄄d呈色为强阳性,细胞形态多为三角形和梭形,胞核不着色,胞质色深。细胞中等大小,以25μm为主。在脊髓颈、胸、腰各段灰质中间带NOS阳性细胞分布的数量基本相等,没有特异性变化。结论肝硬化大鼠与正常大鼠一氧化氮合酶的表达在脊髓内是相同的。NOS在脊髓节段的分布特性可能说明肝硬化大鼠低级交感神经的调控与正常无异。     

一氧化氮合酶阳性神经元在肝硬化大鼠中缝核的分布

目的研究肝硬化大鼠中缝核内一氧化氮合酶阳性神经元分布的变化。方法用四氯化碳建立肝硬化动物模型,NADPH-d组织化学方法观察大鼠脑干中缝核内神经元的变化,图像分析仪对神经元的形态及数量进行分析。结果①NOS阳性神经元及纤维广泛分布于脑干中缝核内,特别是中缝背核,且上段多于下段;②肝硬化大鼠NOS阳性神经元脑干中缝核内分布与正常组基本一致,但阳性反应物的密集度明显增高。结论肝硬化后脑干中缝核一氧化氮合酶较正常明显增多,其分布与兴奋性氨基酸分布相互重叠,可以推测它与一些神经递质在脑内共存并激发神经毒作用。     

大鼠脑干神经元型一氧化氮合酶免疫阳性神经元的分布

目的　观察大鼠脑干神经元型一氧化氮合酶 (nNOS)免疫阳性神经元的分布 ,为探讨nNOS的作用提供形态学资料。方法　用ABC免疫细胞化学方法显示脑干nNOS免疫阳性神经元。结果　大鼠脑干nNOS免疫阳性神经元以中脑和脑桥分布丰富 ,延髓较稀少 ;在中脑 ,nNOS免疫阳性神经元主要分布于中脑水管周围灰质的背侧部、被盖背外侧核、中缝背核、上下丘灰质等部位 ;在脑桥 ,主要分布于被盖背外侧核、脑桥中缝核、被盖脚桥核、蓝斑、臂旁核、斜方体核 ,以及脑桥网状结构 ;与中脑和脑桥相比 ,延髓nNOS免疫阳性神经元较少 ,主要分布于延髓网状结构、三叉神经脊束核和孤束核等核团。结论　分布于脑干内丰富的nNOS免疫阳性神经元可能通过其生成的NO调节其他神经递质的分泌 ,共同参与内脏活动、感觉和运动的传导 ,以及睡眠和觉醒等脑的高级整合功能的调节。     

大鼠下丘脑一氧化氮合酶（NOS）阳性神经元的分布

观察大鼠下丘脑各核团NOS阳性神经元的分布。采用还原型尼克酰胺腺嘌呤二核苷酸脱氢酶（NADPH－d）法，结果显示，大量NOS阳性神经元见于下丘脑外侧区、视上核（SO）和室旁核（Pa）；出现较多NOS阳性神经元的部位是视前大细胞核，见到少量NOS阳性神经元的部位是室周核、视前内侧区、视前外侧区和下丘脑前区。结论：NOS阳性神经元分布于下丘脑的许多核团。     

胰岛素对糖尿病大鼠海马一氧化氮合酶阳性神经元变化的影响

观察实验性糖尿病大鼠海马一氧化氮合酶（NOS）阳性神经元变化及胰岛的治疗作用。给成年SD大鼠腹腔注射链脲佐菌素制备糖尿病大鼠模型，每日给予长效胰岛素2－3U使血糖低于10mmol/L，于第3月及第6月末以NADPH－d组化法显示海马NOS阳性神经元。并做Morris水迷宫行为学测试。海马NOS阳性神经元密度变化如下：糖尿病组齿状回3月时显著减少（P＜0．01），6月时更明显（P＜0．01）；CA1区6月时显著降低（P＜0．01）；治疗组齿状回3月时恢复正常，而6月时低于正常（P＜0．01）但高于糖尿病组3月时（P＜0．05），CA1区3月，6月均恢复正常。行为学测试中各组大鼠逃避潜伏期变化与齿状回NOS阳性神经元密度变化一致。以上变化可能与糖尿病学习记忆功能损伤有关。胰岛素治疗可延缓其发生。     

大鼠纹状体一氧化氮合酶阳性神经元的生后分布及发育

目的：探讨一氧化氮合酶(NOS)阳性神经元在生后不同日龄(1、5、10、15、30、120d)大鼠纹状体中的分布及形态变化特征。方法：NADPH-d组织化学方法。结果：NOS阳性神经元首先出现于尾壳核前上部，随生长发育逐渐扩展至外侧部、内侧部和苍白球；1～5d时胞体直径10μm左右，无突起，或仅1～2个短小突起，10～15d胞体增大，数量增多，突起明显增多伸长，并有许多膨体，30d时胞体直径达20～35μm，突起200μm以上，并反复分支交织成网状，120d与30d比较无明显变化。结论：纹状体内NOS阳性神经元的发育变化在生后1～30d之间，NO可能与运动机能的建立及调控有关。     

哮喘模型大鼠肺组织一氧化氮合酶的分布

研究一氧化氮 (NO)在哮喘大鼠肺组织中的作用。采用组化法观察一氧化氮合酶 (NOS)在大鼠哮喘模型肺组织中的分布 ,应用免疫组化法观察大鼠哮喘模型气道mIL 2R+ 细胞变化。结果显示 ,哮喘大鼠肺NADPH染色呈强阳性 ,并波及肺泡膈。肺组织中NOS含量明显高于对照组 [哮喘组 (37 44± 0 77)pmol/mg,对照组 (8 73± 0 79)pmol/mg],气道炎性细胞增多 ,特别是mIL 2R+ 细胞 [哮喘组mIL 2R+ 细胞为 (2 3 8± 7 9)个 ,对照组为 0个 ],而NOS抑制剂DMA组气道炎性细胞少 ,NADPH呈阴性。提示NO是哮喘大鼠的炎性效应分子。     

一氧化氮合酶阳性神经元及纤维在大鼠脊髓的分布及其来源

用还原型尼克酰胺腺嘌呤二核苷磷酸(NADPH)脱氢酶反应观察了大鼠脊髓内一氧化氮合酶阳性神经元和纤维的分布。胞体浓染且突起显示良好的类似Golgi染色的阳性神经元分布于亘脊髓全长的中央管周围灰质(X层),胸腰髓(T1-L3)的中间带外侧核、中介核及腰骶髓(L6、S1)的骶髓副交感核和后连合核,少量弥散于后角Ⅳ～Ⅵ层。后角Ⅲ层内可见较多的胞体浓染或淡染但突起短或未见阳性突起的小型神经元。前角内无阳性神经元,但有较多串珠状阳性终末支,并在运动神经元胞体和树突上形成终扣。阳性纤维和终末在后角Ⅰ层和Ⅲ层密集,其它部位稀少。在胸腰段和腰骶段脊髓后角内、外侧缘有行向腹侧的内、外侧阳性纤维束,外侧束集中而粗大,阳性纤维束终止于中间带外侧核和后连合核。切断后根和半横断颈髓的实验表明,后角Ⅰ层的阳性纤维和终末主要来源于脊神经节内一氧化氮合酶阳性细胞的中枢突,而Ⅲ层内的阳性纤维和终末则为Ⅲ层和后角深层阳性细胞的突起。结合文献和本文结果可以推断,一氧化氮在躯体和内脏感觉的传入过程及内脏传出活动中是一种重要的活性物质。     

大鼠端脑内一氧化氮合酶阳性神经元的发育

目的研究大鼠胚胎时期及生后早期一氧化氮合酶（NOS）阳性神经元在端脑的分布,探讨一氧化氮（NO）在脑发育过程中的作用。方法应用还原型尼克酰胺腺嘌呤二核苷酸磷酸脱氢酶（NADPH-d）组织化学方法观察孕14d起至生后14d大鼠端脑内NOS阳性神经元的形态和分布。结果孕14d没有观察到阳性神经元。孕15d纹状体腹外侧已有NOS阳性表达。孕17d在大脑皮质、梨状皮质观察到NOS阳性神经元,但胞体小,树突短,且分支少。随着年龄的增长神经元的胞体数目增多、染色增强或维持一定的水平。到孕20d,NOS阳性神经元分布广泛,梨状皮质、纹状体腹外侧及终纹床核均有大量NOS阳性神经元,其胞体明显增大,树突分支复杂化,长度增加。在生后,除上述脑区的阳性神经元进一步发育分化,大脑皮质和纹状体的NOS阳性纤维相互编织成疏密不等的纤维网外,在胼胝体、海马也观察到NOS阳性神经元。到生后14d,NOS阳性神经元的分布模式总体上已与成年大鼠相似。结论NOS阳性神经元在端脑独特的表达模式提示NO在脑发育和成熟过程中扮演重要角色。     

大鼠纹状体内一氧化氮合酶阳性神经元的发育

用还原型尼克酰胺腺嘌呤二核苷酸脱氢酶组化方法观察Ｅ１４ｄ起至生后２８ｄ这段时间大鼠纹状体一氧化氮合酶阳性神经元的发育，结果：胚龄１６ｄ时，尾壳核内已有一氧化氮合酶的表达，此时细胞数量少，细胞圆形，无突起或仅有一短的突起，胚龄１８ｄ一氧化氮合酶阳性神经元的数量增多，生后１display status     

一氧化氮合酶(NOS)与子宫内膜关系的研究进展

女性子宫内膜在女性生殖生理功能方面担负着月经形成、胚胎着床、妊娠维持、激素分泌等重要作用。自从1995年Telfer首次发现人子宫内膜中NOS(N itric Oxide Synthase)mRNA和其蛋白的存在后,近年来越来越多的研究表明NO(N itric Oxide)在女性生殖过程中扮演着重要的角色。本文将就NOS在子宫内膜的活性表达,及其与子宫内膜关系的研究作一综述。     

Effect of focal cerebral ischemia on nitric oxide synthase expression in rats

Time- and cell-type-dependent immunohistochemical activity of nitric oxide synthase (NOS) was investigated in rat cerebral cortex following focal ischemia and the local concentration of nitric oxide (NO) was measured. NO concentration increased 2 min after the ischemia. Brain NOS-immunoreactive neurons increased in number 5 min after the ischemia. Endothelial cell NOS immunoreactivity was first detected in vascular endothelial cells and astrocytes 5 min after the ischemia, and it increased again during 60 min to 4 days after the ischemia in reactive astrocytes. Inducible NOS immunoreactivity was detected in astrocytes, vascular endothelium, and microglia/macrophages at the periphery of the ischemic core during 2–4 days after the ischemia.This study was presented at the 28th Annual Meeting of the Clinical Electron Microscopy Society of Japan, Osaka, October 17–19, 1996.     

Expression of nitric oxide synthase (NOS) genes in channel catfish is highly regulated and time dependent after bacterial challenges

Nitric oxide is well known for its roles in immune responses. As such, its synthesizing enzymes have been extensively studied from various species including some teleost fish species. However, the NOS genes have not been characterized in channel catfish (Ictalurus punctatus). In this study, we identified and characterized three NOS genes including one NOS1 and two NOS2 genes in channel catfish. Comparing with the NOS genes from other fish species, the catfish NOS genes are highly conserved in their structural features. Phylogenetic and syntenic analyses allowed determination of NOS1 and NOS2 genes of channel catfish and their orthology relationships. Syntenic analysis, as well as the phylogenetic analysis, indicated that the two NOS2 genes of catfish were lineage-specific duplication. The NOS genes were broadly expressed in most tested tissues, with NOS1 being expressed at the highest levels in the brain, NOS2b1 highly expressed in the skin and gill, and NOS2b2 lowly expressed in most of the tested tissues. The most striking findings of this study was that the expression of the NOS genes are highly regulated after bacterial infection, with time-dependent expression patterns that parallel the migration of macrophages. After Edwardsiella ictaluri challenge, dramatically different responses among the three NOS genes were observed. NOS1 was only significantly in the skin early after infection, while NOS2b1 was rapidly upregulated in gill, but more up-regulated in trunk kidney with the progression of the disease, suggesting such differences in gene expression may be reflective of the migration of macrophages among various tissues of the infected fish. In contrast to NOS1 and NOS2b1, NOS2b2 was normally expressed at very low levels, but it is induced in the brain and liver while significantly down-regulated in most other tissues.     

Fibre-related nitric oxide synthase (NOS) in Duchenne muscular dystrophy

Nitric oxide (NO) mediates fundamental physiological actions on skeletal muscle. The loss of NO synthase (NOS) from the sarcolemma was assumed to be associated with development of Duchenne muscular dystrophy (DMD). We have, however, recently reported that, in contrast to the commonly accepted view, NOS expression in DMD myofibres is up-regulated. This poses the question of the fibre type-specific NOS expression in DMD muscles and how the NOS expression is related to the regeneration or degeneration status. To address this issue, we examined localization of NOS isoforms I, II and III in skeletal muscles of DMD patients employing immunohistochemical labelling with tyramide signal amplification complemented with enzyme histochemistry. We found that NOS immunolabelling as well as metabolic enzyme activity in DMD muscles were heterogeneously distributed along the fibre length of DMD muscle fibres revealing regenerating and degenerate (hypercontracted) fibres as well as normal segments. Like in normal muscles, positive NOS immunoreactivity was found to be associated with fast-oxidative glycolytic (FOG) phenotype. The regeneration status of NOS-positive segments was deduced from the presence of neonatal and developmental myosin heavy chains. High NOS expression in regenerating DMD muscle fibres can be well reconciled with reports about the protective role of endogenous NO in inflammatory diseases and in muscle repair.     

扬子鳄食管颈段一氧化氮合酶(NOS)阳性神经元及神经纤维的分布

本实验采用还原型尼克酰胺腺嘌呤二核苷酸脱氢酶(NADPH-d)方法,观察扬子鳄食管颈段NOS阳性神经元和神经纤维的分布.结果食管颈段的粘膜下层和肌层分布着较丰富的NOS阳性神经纤维,一些神经纤维交织成丛状,其间散在分布着NOS阳性神经元胞体.粘膜下一些NO S阳性神经丛环绕血管壁走行.     

一氧化氮/诱导型一氧化氮合酶在动脉粥样硬化过程中的作用

 目的：探讨一氧化氮（NO）/诱导型一氧化氮合酶（iNOS）在动脉粥样硬化（atherosclerosis,AS）过程中的动态变化,分析其对动脉粥样硬化形成过程的影响。方法：将60只SD大鼠随机分成2组：对照组及AS组,每组30只。AS组采用维生素D3腹腔注射联合高脂饲料饲养的方法构建动脉粥样硬化模型。用相关生化方法检测血清各项生化指标：总胆固醇、甘油三酯、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、空腹血糖和钙离子,比色法检测血清NO浓度,并对主动脉行HE染色,免疫组化技术检测iNOS蛋白表达,将所得数据进行统计分析,用简单线性相关分析NO与钙离子及动脉粥样硬化指数的相关性。结果：90 d后成功构建了主动脉中膜钙化型动脉粥样硬化模型。血清NO浓度在动脉粥样硬化过程中逐步下降,各组间差异均有统计学意义（均P<0.05）。动脉粥样硬化过程中动脉粥样硬化指数与钙离子呈正相关,与NO呈负相关。在90 d的AS组粥样斑块区免疫组化技术检测到iNOS蛋白表达。结论:在动脉粥样硬化形成过程中,主动脉粥样斑块区iNOS蛋白高表达,但血清NO浓度逐渐降低,NO抗动脉粥样硬化作用减弱。     

The expression and activity of renal nitric oxide synthase and circulating nitric oxide in polycystic kidney disease rats

Nitric oxide (NO) influences tubular fluid and electrolyte transport, and hence possibly also fluid accumulation in renal cysts. The expression and activity of intrarenal constitutive NO synthase (cNOS) [neuronal NOS, nNOS and endothelial NOS, eNOS] and inducible NOS (iNOS) and plasma nitrite/nitrate (PNOx) concentration were assessed in homozygous Han:SPRD polycystic kidney disease (PKD) rats (cy/cy), heterozygous Han:SPRD PKD rats (cy/+), homozygous normal Han:SPRD littermates (+/+) and Sprague Dawley rats (sd). The results showed: 1) nNOS expression was decreased in proximal tubules and thick ascending limbs of the loop of Henle in cy/cy and cy/+ rats compared to +/+ and sd rats (p<0.05). nNOS was weakly expressed in the epithelium of small cysts and unexpressed in epithelium of large cysts. 2) iNOS expression was increased in proximal tubular epithelial cells in cy/+ rats compared to +/+ rats and sd rats (p<0.01). iNOS expression in cyst epithelium was decreased in cy/+ rats (p<0.05) and absent in cy/cy rats. 3) eNOS expression was similar in the endothelium of intrarenal arteries in all groups. 4) The activity of renal cNOS was decreased in cy/cy and cy/+ rats; the activity of iNOS was decreased only in cy/cy rats, with no significant difference among the other three groups. 5) PNOx concentration was higher in cy/cy rats than in the other three groups, and correlated positively with plasma creatinine and urea. In conclusion, NOS expression and activity decreased as cysts developed, suggesting that NO downregulation is involved in the pathogenesis of PKD.