一氧化氮和一氧化氮合酶在鼠脑内形成吗啡依赖的作用研究

目的：研究一氧化氮（NO）和一氧化氮合酶（NOS）在吗啡依赖形成中的作用。方法;对吗啡依赖和戒断大鼠脑内NO含量和NOS活力进行测定。结果：未发现吗啡依赖和戒断大鼠脑内NO含量和NOS活力有改变。结论：对NO／NOS与吗啡依赖的关系还有待进一步研究。     

大剂量伽玛刀照射后脑组织一氧化氮合酶亚型表达改变及其与急性脑水肿的关系

目的探讨大剂量伽玛刀(γ刀)照射后脑组织一氧化氮合酶(NOS)亚型表达改变及其与急性脑水肿的关系。方法200Gy量γ刀照射正常大鼠脑,采用光镜、电镜、免疫组化及原位杂交技术研究照射后急性脑水肿的发生发展及脑组织NOS亚型表达变化。结果①照射后30min出现急性脑水肿病理改变,照射后2h出现明显血管源性脑水肿,照射后6h出现明显细胞性脑水肿,照射后3d急性脑水肿达高峰。②脑组织NOS亚型表达在照射后30min开始增高,内皮型一氧化氮合酶(eNOS)及诱导型一氧化氮合酶(iNOS)表达分别于照射后2h及6h显著增高,神经元型一氧化氮合酶(nNOS)表达亦增高,但nNOS阳性表达细胞数量较少。3种NOS亚型表达增高持续时间长,伴行于脑水肿的急性发展阶段。结论大剂量γ刀照射后脑组织NOS亚型表达增高与急性脑水肿的发生发展有关。     

海人酸致痫动物模型脑内一氧化氮,一氧化氮合酶的变化

目的探讨一氧化氮（ＮＯ）、一氧化氮合酶（ＮＯＳ）在癫痫发生中的作用及ＮＯＳ抑制剂的作用。方法采用海人酸致痫大鼠模型并应用ＮＯＳ抑制剂Ｌ－硝基精氨酸甲酯（Ｌ－ＮＡＭＥ）,分别在致痫后３０分钟、６０分钟取海马组织,匀浆后测定ＮＯ及ＮＯＳ水平。结果致痫３０分钟后海马ＮＯ含量显著升高,至６０分钟恢复正常;ＮＯＳ活性水平增高＞５０％;Ｌ－ＮＡＭＥ明显抑制大鼠的痫性发作,应用ＮＯＳ抑制剂组大鼠海马ＮＯ、ＮＯＳ含量明显下降。结论癫痫发作后脑内ＮＯ、ＮＯＳ活性增强,ＮＯＳ抑制剂通过抑制酶活性使ＮＯ生成降低,并完全抑制痫性发作。ＮＯＳ活性受抑制＞４８％即可产生明显效果。提示ＮＯ可能有内源性致痫作用。     

一氧化氮和一氧化氮合酶在大鼠脑损伤组织中的表达

目的 探讨创伤性脑损伤(traumatic brain injury,TBI)后脑组织匀浆中一氧化氮(nitric oxide,NO)含量、一氧化氮合酶(nitric oxide synthase,NOS)活性以及与脑水肿之间的关系.方法 SD大鼠52只,36只随机分为正常对照组(n=6)和手术组(n=30),手术组按受伤到处死的不同时间(6、24、72、120、168 h)分成5个亚组,每亚组6只,各组取脑组织匀浆检测NO、NOS及测定脑组织含水量.16只随机分4组,正常对照组、伤后6h、72 h、168 h各4只,行尼克酰胺腺嘌呤二核苷酸磷酸黄递酶(Nicoti-namide adenine dinucleotide phosphate diaphorase,NADPHd))组化染色检测皮层及脑底NOS阳性细胞.结果:TBI后脑组织内NO含量、NOS活力即有升高,与对照组比较有明显差异(NO含量比较F=468.89,NOS活力比较F=84.32,P＜ 0.05).脑组织含水量在外伤后升高,与对照组比较有明显差异(F=1963.51,P＜0.05).NADPHd组化染色显示TBI皮层NOS阳性细胞明显多于正常对照组,伤灶脑底也出现了染色块及浓染的细胞群与阳性纤维束.结论 大鼠TBI后损伤灶外确实发生了NO、NOS的升高,伤后1周内持续存在;NO含量、NOS活性与脑组织含水量的变化趋势基本一致,为临床治疗脑水肿提供实验依据.     

癫痫发作后脑内一氧化氮、一氧化氮合酶及相应氨基酸的变化

目的　探讨NO在癫痫的病理生理过程中的作用及可能作用机制。方法　将大鼠随机分为对照组 ,实验性癫痫组 ,L 硝基精氨酸甲酯 (L NAME)抑制组 ,分别取双侧海马及颞叶皮层组织测定NO ,NOS及相应氨基酸的含量。结果　致痫后NOS活性上升 >50 %。NO含量明显上升 ,60分钟组NO含量明显下降。L Arg,Glu含量均显著上升。GABA则明显下降。不同剂量的L -NAME均抑制痫性发作及相应指标的变化。癫痫患者脑脊液中NO ,NOS含量均较对照组明显上升。结论　NO参与癫痫发生的机制 ,但对其维持作用并不重要。其致痫机制仅部分与兴奋性氨基酸有关。     

癫痫患儿血清一氧化氮和一氧化氮合酶含量的变化及意义

目的　探讨癫疒间 患儿血清一氧化氮 (NO)、一氧化氮合酶 (NOS)的变化及意义。方法　利用ELISA方法 ,测定 5 8例癫疒间 患儿 (癫疒间 组 )和 2 3名健康儿童 (对照组 )血清中NO、NOS的含量 ,并分组比较不同条件下其含量的变化。结果　癫疒间 组血清NO、NOS的含量分别为 (5 .86± 1.2 1) μmol/ml和 (2 8.2 6± 8.4 9)U/ml,较对照组的 (3.78± 0 .74 ) μmol/ml及 (17.86± 4 .5 8)U/ml明显升高 (P <0 0 1) ;发作近期为 (7.31± 1.2 7)μmol/ml和 (31.2 5± 11.35 )U/ml,明显高于发作间期 (4 .2 7± 0 .6 6 ) μmol/ml和 (2 4 .15± 7.85 )U/ml(P <0 0 1) ;癫疒间 组EEG异常者为 (7.18± 1.35 ) μmol/ml和 (34.4 8± 8.5 6 )U/ml,明显高于EEG正常者 (4 .0 4± 0 .75 ) μmol/ml和 (2 2 .85± 7.4 5 )U/ml(P <0 0 1) ;但与发作类型、病程及是否接受治疗无关 (P >0 0 5 )。结论　癫疒间 发作近期血中NO、NOS生成增加 ,NO作为内源性调质参与癫疒间 发作病理生理过程     

Aβ1—40海马注射对大鼠脑内一氧化氮合酶表达的影响

目的探讨一氧化氮合酶(NOS)在β淀粉样蛋白(Aβ)神经毒性及阿尔茨海默病(AD)病理机制中的作用.方法应用免疫组化方法,观察大鼠海马齿状回Aβ1-40注射后神经元型一氧化氮合酶(nNOS)和诱导型一氧化氮合酶(iNOS)表达变化.结果正常大鼠海马齿状回区含nNOS神经元计数为8.96±0.35个/视野;生理盐水注射后局部含nNOS神经元无明显变化(8.97±0.29个/视野);Aβ1-40注射后,注射区周围含nNOS神经元数目显著减少(2.98±0.24个/视野).正常及生理盐水注射组脑内未见iNOS表达;Aβ1-40注射后2d、10d和30d,注射区持续出现大量含iNOS的胶质细胞(主要为星形胶质细胞),反应面积分别为0.905±0.082、0.962±0.161、0.935±0.125mm2.结论Aβ1-40海马注射可损伤局部含nNOS神经元及诱导胶质细胞iNOS表达,NOS在Aβ神经毒性和AD发病中有重要作用.     

一氧化氮／一氧化氮合酶与神经创伤

一氧化氮是一种简单的气体分子,可在哺乳类神经细胞内经一氧化氮合酶作用产生。ＮＯ在神经创伤修复中的多重作用近年来已受到越来越多的重视。本文对ＮＯ／ＮＯＳ与神经创伤和再生之间的关系作一综述。     

二硫化碳对大鼠海马一氧化氮合酶活力和基因表达的影响

目的:探讨二硫化碳(CS2)对大鼠海马一氧化氮合酶(NOS)活力和基因表达的影响。方法:以吸入染毒法制作不同浓度CS2中毒大鼠模型:染毒2个月后,用Morris水迷宫法检测实验大鼠的学习记忆功能;以NOS测定试剂盒测定大鼠海马NOS活力;以半定量逆转录一聚合酶链式反应(RT-PCR)法测定神经元型一氧化氮合酶(nNOS)mRNA 含量的变化。结果:Morris水迷宫测试显示染毒后大鼠平均逃逸潜伏期较对照组延长,差异有显著性;各CS2染毒组大鼠海马NOS活性降低,与对照组比较差异有显著性,随CS2浓度的增加NOS活性降低,各染毒组间比较差异有显著性差异;染毒后海马nNOS mRNA含量比对照组显著减少,与CS2浓度呈剂量依赖关系,差异有显著性意义。结论:CS2致大鼠海马NOS活力降低及nNOS mRNA含量减少可能是CS2干扰学习记忆功能的机制之一。     

一氧化氮和一氧化氮合酶在大鼠局灶性脑缺血中的表达特点

目的建立脑缺血SD大鼠模型,探讨一氧化氮(NO)和一氧化氮合酶(NOS)在脑缺血模型中的表达特点。方法应用线栓法制作大脑中动脉阻塞(MCAO)局灶性脑缺血模型,根据缺血不同时间分为8组,设立假手术组和正常对照组,每组各有6只大鼠。应用硝酸还原酶法测定脑组织NO的含量,流式细胞术(FCM)定量检测硝基酪氨酸(NT)表达,化学比色法测定脑组织NOS的活性,免疫组织化学方法定位检测eNOS、nNOS和iNOS表达位置,逆转录反应系统(RT-PCR)半定量分析eNOS、nNOS和iNOS的 mRNA在脑缺血区域的表达。结果神经功能缺失评分发现缺血时间越长,神经功能缺失越明显;脑组织中NO含量与缺血时间正相关;缺血1h后NT阳性细胞百分比开始明显升高(9.50%);缺血0.5h时NOS的活性开始升高,缺血3d达到高峰[0.94nmol/(g.min)];免疫组织化学提示eNOS在神经细胞和血管内皮细胞胞浆均有表达,nNOS和iNOS抗体主要在神经细胞胞浆中表达;RT-PCR半定量分析在缺血早期(0.5h~6h),随着缺血时间延长,eNOS和nNOS表达增加,iNOS未见表达或低表达;缺血中晚期(6h~5d),iNOS高表达,并与缺血时间呈正相关,eNOS和nNOS表达明显减少。结论脑缺血时间延长,NOS的活性升高,NO在体内特异性代谢产物NT量增加,神经功能缺失越明显;NOS在缺血早期以eNOS和nNOS为主,在缺血晚期以iNOS为主。     

Interventional effect of magnesium sulfate on nitric oxide synthase activity after acute craniocerebral injury

BACKGROUND: Abnormal changes in magnesium ion are closely related to cerebral injury. At present, some evidence indicates that magnesium reagent can improve nerve function and prognosis of patients with cerebral injury. OBJECTIVE: To observe the effect of magnesium sulfate on changes in nitric oxide synthase (NOS) activity in brain tissue of rats with acute craniocerebral injury. DESIGN: Completely randomized grouping design and randomly controlled study. SETTING: Laboratory of Neurosurgery, the Third Hospital of Chinese PLA. MATERIALS: Fifty-four male SD rats of clean grade and weighing 220–250 g were randomly divided into normal control group (n =6), cerebral injury group (n =24) and magnesium sulfate group (n =24). Especially, rats in cerebral injury group and magnesium sulfate group were equally divided into four subgroups and observed at 0.5, 2, 6 and 24 hours after model establishment. A solution of 125 g/L of magnesium sulfate was provided by the Seventh Pharmaceutical Factory of Wuxi and the NOS assay kit by Nanjing Jiancheng Bioengineering Institute. METHODS: The experiment was carried out in the Institute of Neurosurgery, the Third Hospital of Chinese PLA from August 2000 to August 2002. ① Rats in the cerebral injury group and magnesium sulfate group were anesthetized to establish cerebral injury models based on modified Feeney technique; magnesium sulfate group were intraperitoneally injected 600 mg/kg magnesium sulfate (125 g/L), but rats in the normal control group remained untreated. ② At 0.5, 2, 6 and 24 hours after cerebral injury, rats in cerebral injury group and magnesium sulfate group were decapitated and brains were dissected. Cerebral cortex of rats in cerebral injury group was selected for NOS assay; in addition, at 0.5 hour after cerebral injury, a portion of the parietal lobe was selected from the brains of rats in the normal control group. Brain samples were homogenized, the homogenated centrifuged and the supernatants were used to measure NOS activity with NOS kit. ③ Differences among the three groups were compared with t test. MAIN OUTCOME MEASURES: NOS activity in cerebral cortex of rats in each group. RESULTS: A total of 54 SD rats were involved in the final analysis. At 0.5 hour after cerebral injury, NOS activity in cerebral cortex was (42.45±13.46) nmol/L in cerebral injury group and (41.17±12.53) nmol/L in magnesium sulfate group, respectively, which was higher than that in normal control group [(39.45±11.84) nmol/L, P < 0.05]. At 2 hours after cerebral injury, NOS activities were (66.48±21.43) and (63.24±19.18) nmol/L, respectively, while at 6 hours after cerebral injury, NOS activities were (62.45±24.18) and (51.97±20.46) nmol/L, respectively, which were higher than those in normal control group (P < 0.01). At 24 hours after cerebral injury, NOS activity returned to basal level. Moreover, NOS activity was significantly lower in the magnesium sulfate group than that in the cerebral injury group at 2 and 6 hours after cerebral injury (P < 0.05, 0.01). CONCLUSION: NOS activity is increased in injured brain tissue of rats with craniocerebral injury, and treatment with magnesium sulfate provides some degrees of protection possibly through inhibition of NOS activity after cerebral injury.     

糖皮质激素干预肺源性脑损伤大鼠血清一氧化氮合酶活性和一氧化氮含量的变化

实验观察地塞米松及Gi蛋白抑制剂百日咳毒素,转录抑制剂放线杆菌素干扰后肺源性脑损伤大鼠模型大鼠血清一氧化氮合酶活性,一氧化氮含量变化。 发现高剂量地塞米松(13 mg/kg)和地塞米松联合放线菌干预肺源性脑损伤模型大鼠肺含水量降低,血清一氧化氮合酶活性和一氧化氮含量明显增高,肺含水量降低,肺泡间质内炎细胞浸润减少,脑膜血管充血明显减轻,浸润的胶质细胞较少。结果说明,高剂量糖皮质激素可通过提高血清一氧化氮合酶活性,增加一氧化氮含量在肺源性脑损伤中发挥保护作用。     

Inhibition of nitric oxide synthase aggravates brain injury in diabetic rats with traumatic brain injury

Studies have shown that hyperglycemia aggravates brain damage by affecting vascular endothelial function. However, the precise mechanism remains unclear. Male Sprague-Dawley rat models of diabetes were established by a high-fat diet combined with an intraperitoneal injection of streptozotocin. Rat models of traumatic brain injury were established using the fluid percussion method. Compared with traumatic brain injury rats without diabetic, diabetic rats with traumatic brain injury exhibited more severe brain injury, manifested as increased brain water content and blood-brain barrier permeability, the upregulation of heme oxygenase-1, myeloperoxidase, and Bax, the downregulation of occludin, zona-occludens 1, and Bcl-2 in the penumbra, and reduced modified neurological severity scores. The intraperitoneal injection of a nitric oxide synthase inhibitor N(5)-(1-iminoethyl)-L-ornithine(10 mg/kg) 15 minutes before brain injury aggravated the injury. These findings suggested that nitric oxide synthase plays an important role in the maintenance of cerebral microcirculation, including anti-inflammatory, anti-oxidative stress, and anti-apoptotic activities in diabetic rats with traumatic brain injury. The experimental protocols were approved by the Institutional Animal Care Committee of Harbin Medical University, China(approval No. ky2017-126) on March 6, 2017.     

一氧化氮合酶活性与抑郁症的相关性

目的通过对抑郁症患者一氧化氮合酶（NOS）活性进行检测，从而研究和探讨一氧化氮合酶、一氧化氮（NO）与抑郁症之间的关系。方法采用分光光度法检测抑郁症患者治疗前后的一氧化氮合酶NOS及其亚型（结构型cNOS、诱导型iNOS）的活性，并与正常对照组比较。结果抑郁症组的NOS、cNOS活性显著低于正常对照组；治疗组的NOS、cNOS活性高于抑郁症（无显著性），但治疗后缓解组的NOS、cNOS活性均显著高于治疗前。各组iNOS的活性无显著差异。结论抑郁症病人的NOS活性下降，而且主要是结构型cNOS活性下降，经治疗缓解后有所提高。因此，NOS和NO很有可能在抑郁症的发病过程中起着重要作用。     

麝香注射液对脑出血后脑水肿的影响及临床疗效

目的:探讨麝香注射液对脑出血后脑水肿的影响及临床疗效。方法:高血压脑出血患者98例,随机分为麝香注射液治疗组(45例)和常规治疗组(53例),所有患者均于治疗前及治疗两周后行头颅CT扫描,并用美国国立卫生研究院卒中量表(NIHSS)进行神经功能评分,用Barthel指数进行日常生活活动(ADL)能力评分。结果:治疗两周后,麝香注射液治疗组水肿体积、NIHSS评分及ADL能力评分的改善显著优于常规治疗组。结论:麝香注射液有助于控制和减轻脑出血后脑水肿,提高临床疗效。     

蛋白酶体抑制剂诱导多巴胺能神经元变性及诱导型一氧化氮合酶变化

目的探讨蛋白酶体（proteasome）功能下降在帕金森病（PD）发病机制中的作用，以及模型大鼠脑内黑质部位诱导型一氧化氮合酶（iNOS）是否参与蛋白酶体抑制剂Lactacystin诱导的多巴胺能神经元变性。方法将30只健康雄性SD大鼠分为5组（生理盐水对照组，1d组、3d组、1周组、3周组），每组6只。将蛋白酶体抑制剂Lactacystin立体定向注射至大鼠黑质部位，记录大鼠在不同时间点的行为学改变，并用免疫组化方法观察生理盐水对照组及不同时间点组（1d、3d，1周、3周）大鼠黑质区多巴胺能神经元变性及iNOS变化。结果Lactacystin注射1周后大鼠开始出现自发性活动减少，阿朴吗啡可诱导出旋转行为；3周后，30min旋转次数为258．90±11．56；实验3周组黑质部位TH阳性细胞减少。1d后iNOS阳性细胞明显增多，3d时达高峰，1周后开始下降，3周时基本消失。结论蛋白酶体功能下降可能是多巴胺能神经元变性的始动因素，而iNOS上调可能是多巴胺能神经元变性的重要过程。     

磁共振成像评估丁苯酞软胶囊对创伤性脑水肿大鼠的治疗作用

目的运用磁共振成像探索大鼠创伤性脑水肿的演变规律及丁苯酞软胶囊对其的治疗作用。方法 SD雄性大鼠175只。随机分为非干预组、干预组和正常对照组。采用Marmarou法制作创伤性脑水肿大鼠模型,造模成功后分别于6 h、12 h、1 d、3 d、7 d行磁共振成像扫描、HE染色、脑组织含水量测定、伊文思蓝渗出量测定。结果磁共振成像、HE染色、和脑组织含水量测定结果提示与对照组相比,非干预组和干预组脑组织可见明显脑水肿,其中以损伤后24 h最明显,3 d后开始减轻。干预组较非干预组脑水肿程度明显减轻。伊文思蓝渗出检测提示,创伤性脑损伤后伊文思蓝渗出量增多,其中以6 h渗出量最多。非干预组较干预组渗出明显。结论创伤性脑损伤后,继发血管源性脑水肿和细胞毒性脑水肿,其中以1 d时脑水肿最明显,丁苯酞软胶囊能明显减轻脑损伤后脑水肿的形成。     

Expression of nitric oxide synthase in the cerebral microvasculature after traumatic brain injury in the rat

Reduced nicotinamide adenine dinucleotide phosphate-diaphorase (NADPHd) histochemistry and nitric oxide synthase (NOS) immunocytochemistry were performed on sections of brain after moderate traumatic brain injury. There was a pronounced increase in NADPHd reactivity and an induction of the endothelial NOS (eNOS) isoform in microvessels surrounding the cortical contusion by 24 h post-injury. This altered microvascular state may contribute to barrier breakdown and hyperemia which characterize traumatic brain injury.     

Involvement of nitric oxide and nitric oxide synthase activity in anticonvulsive action

The anticonvulsant drug Diazepam (DIA-2 mg/kg b. wt), the nitric oxide (NO) donor L-Arginine (L-Arg-2000 mg/kg b. wt) and the putative nitric oxide synthase (NOS) inhibitor N(G)-Nitro-L-Arginine methyl ester (L-NAME-50 mg/kg b. wt) were used to determine the role of endogenous NO on convulsions induced by picrotoxin (PCT-5 mg/kg b. wt) in rats. Rats given a convulsant dose of PCT (5 mg/kg b. wt) had convulsion and it suppresses the NOS activity and NO concentration in brain regions. The anticonvulsant L-Arg alone significantly increases the NO concentration and NOS activity in brain regions, but not diazepam. Whereas DIA, along with L-Arg, enhances the NO and NOS activity when compared to L-Arg alone. The combination of both OIA and L-Arg completely suppressed the convulsions. L-NAME alone had no effect to produce convulsions but it completely decreased NO concentration and NOS activity and potentiated the PCT convulsions. This was reverted by pre- and post treatment of DIA plus L-Arg indicating, the increased NO concentration and NOS activity in brain regions suppresses convulsions.