REGULATION OF RAT NEURONAL NITRIC OXIDE SYNTHASE ACTIVITY BY CHRONIC ALCOHOLIZATION

Rat neuronal nitric oxide (NO) synthase (nNOS) activity wasmeasured in frontal cortex, hippocampus, striatum and cerebellumusing the assay of [³H]citrulline, following chronic alcoholization.The K_m and V_max values were significantly increased in the frontalcortex and in the striatum, and were not affected in the cerebellumand hippocampus.     

THE EFFECTS OF THE NITRIC OXIDE SYNTHASE INHIBITOR 7-NITROINDAZOLE ON ETHANOL PHARMACOKINETICS IN RATS AFTER ACUTE AND CHRONIC ETHANOL ADMINISTRATION

The aim of this work was to study the effects of the nitricoxide synthase (NOS) inhibitors 7-n,troindazole (7-NI) and N^G-nitro-L-arginine(L-NOARG) on the effects and pharmacokinetics of ethanol inrats. Ethanol at a dose of 4 g/kg, i.p. induced sleep in rats(sleep time: 117.2 ± 30.7 min). Administration of theNOS inhibitors 7-NI (20 mg/kg, i.p.) and L-NOARG (20 mg/kg,i.p.) 30 min before ethanol significantly increased the durationof ethanol-induced sleep. L-NOARG also significantly increasedthe toxicity of ethanol as evidenced by increased post-experimentallethality Ethanol at a dose of 2 g/kg (i.p.) did not inducesleep in vehicle-treated rats; however, the combined administrationof ethanol (2 g/kg) and 7-NI at doses of 40, 80, and 120 mg/kgcaused sleep, for 49.4 ± 3.7, 204.0 ± 13.3, and447.5 ± 62.8 min, respectively. L-NOARG (20 mg/kg) hadno effect on ethanol concentrations in blood after acute ethanoladministration (4 g/kg). 7-NI in lower doses (20 and 40 mg/kg)had no effect and in higher doses (80 and 120 mg/kg) significantlyslowed ethanol clearance during the 12 h after ethanol administration.The effect of 7-NI (20 mg/kg) on ethanol pharmacokinetics afterchronic ethanol administration (inhalation for 18 days) wasalso studied. The administration of 7-NI immediately after theend of ethanol exposure had a pronounced effect on ethanol pharmacokinetics;in 7-NI-treated rats the fall in ethanol concentrations wassignificantly slower as compared with vehicle-treated rats.In 7-NI- treated rats, blood-ethanol levels were higher at 3,6, 9, and 12 h after the end of ethanol exposure.     

THE INFLUENCE OF CHRONIC MODERATE ETHANOL ADMINISTRATION ON NADPH-DIAPHORASE (NITRIC OXIDE SYNTHASE) ACTIVITY IN RAT BRAIN

Nitric oxide synthase (NOS), the enzyme with reduced nicotinamide-adeninedinucleoude phosphate (NADPH)-diaphorase activity, generatesnitric oxide (NO) which is an important bioregulatory moleculein the nervous, immune, and cardiovascular systems. NOS is linkedto non-adrenergic non-cholinergic (NANC) neuronal pathways andmodulation of the N-methyl-D-aspartate receptors, yet its modificationby ethanol has been little explored. A possible modificationby chronic ethanol administration of activity and*or localizationof NADPH-diaphorase (NO-synthase) in rat brain may thus providethe pathogenic basis of alcohol-induced brain injury. When femaleWistar rats were treated chronically with ethanol for 50 days,the NADPH-diaphorase staining of granular neurons and neuronslocated in the molecular layer of the cerebral cortex was significantlyreduced. Chronic ethanol consumption led to a significant reductionin NADPH-diaphorase staining in the superficial layers of thesuperior colliculus. The number of NADPH-diaphorase-positiveneurons was significantly reduced (P < 0.001) in the stratumzonale and stratum griseum superficiale (by 42 3–65 6%of control values). This could alter synaptic processes in thehighly organized structures involved in oculomotor and somaticmotor coordination and thus contribute to the motor disturbanceswhich are associated with alcohol abuse.     

锌对染铅大鼠海马一氧化氮合酶的保护作用

目的 :　探讨铅对海马长时程增强 (LTP)的影响与海马不同亚区一氧化氮合酶(NOS)变化的关系以及锌的拮抗作用。方法 :　采用反映学习记忆功能的 Y迷宫法测试大鼠神经行为的改变 ;用 NADPH-黄递酶 (NADPH- d)组化法和神经元型 NOS(n NOS)的免疫组化法研究大鼠海马不同亚区 NOS的活性及表达情况。结果 :　染铅组大鼠的学习记忆能力比铅锌组和对照组明显下降 (P<0 .0 5 ) ,铅锌组与对照组之间无差别 ;组化及免疫组化显示 :染铅组大鼠海马CA1区和齿状回的 NOS和 n NOS阳性神经元明显少于铅锌组和对照组 (P<0 .0 5 ) ,在 CA3区无差别 ,铅锌组与对照组各亚区均无差别。结论 :　铅可损伤大鼠学习记忆能力 ,鉴于 NOS参与LTP这一代表学习记忆的电生理指标的形成和维持 ,推测铅对学习记忆和海马 LTP的影响可能与染铅后海马各区 NOS的不同变化有关。锌对铅引起的学习记忆损伤和 NOS的影响有拮抗作用。     

冷暴露大鼠血浆肾上腺素及血管内皮细胞 NOS活性的变化

目的观察冷暴露大鼠血浆肾上腺素(Adr)水平及血管内皮细胞NOS活性的变化,为探寻防治冷损伤的措施提供理论依据。方法使用L-精氨酸(L-Arg)调节主动脉内皮细胞NOS活性。大鼠随机分为5组,即对照组(室温)、冷暴露Ⅰ组(-15℃,1 h)、冷暴露Ⅱ组(-15℃,1.5 h)、L-Arg组(室温,L-Arg 2 g.kg-1灌服)、冷暴露 L-Arg组(-15℃,1 h;L-Arg 2 g.kg-1灌服)。冷暴露后6 h再取血样。以酶联免疫法测定血浆肾上腺素水平;以硝酸酶还原法测定血管内皮细胞NOS活性;以紫外分光光度计检测血清和血管内皮细胞培养液中的LDH活性;反转录PCR方法检测血管内皮细胞NOS mRNA表达水平。结果对照组、冷暴露Ⅰ组与Ⅱ组血浆中Adr分别为(10.81±1.85)、(31.37±4.48)、(39.35±5.59)nmol.L-1,冷暴露大鼠血浆中Adr水平明显地高于对照组(P<0.05);上述3组血管内皮细胞NOS活性分别为(16.13±3.68)、(9.19±1.87)、(5.94±1.05)μmol.L-1;冷暴露大鼠血管内皮细胞NOS活性明显低于对照组(P<0.05),使用L-Arg可使冷暴露大鼠血管内皮细胞NOS活性上调到(12.87±1.60)μmol.L-1(P<0.01)。同时,血浆中LDH水平也明显降低(P<0.01)。结论冷暴露血浆高浓度肾上腺素可抑制主动脉内皮细胞NOS活性;L-Arg可上调血管内皮细胞NOS活性,对减轻机体冷损伤程度有一定作用。     

Caveolin-1与内皮型一氧化氮合成酶在门静脉高压症性血管病变中的表达

目的检测肝硬化门静脉高压症病人的脾血管中小凹蛋白caveolin-1、内皮型一氧化氮合成酶(endothelial nitric oxide synthase,eNOS)的mRNA及蛋白表达水平的变化,探讨caveolin-1的表达对eNOS的影响。方法逆转录-聚合酶链反应(RT-PCR)方法检测27例肝硬化门静脉高压症病人脾静脉组织和20例脾外伤病人正常脾静脉血管中caveolin-1和eNOS的mRNA;Western Blot检测caveolin-1和eNOS的蛋白表达水平变化。结果肝硬化门静脉高压症组脾静脉Caveolin-1 mRNA与对照组脾静脉组织表达分别为(0.73±0.18)、(0.38±0.12),两组比较差异有显著性(p<0.05);肝硬化门静脉高压症组脾静脉eNOS mRNA为(0.23±0.11),显著低于对照组内脾静脉组织eNOS mRNA的表达(0.47±0.15)(p<0.05)。Western Blot检测caveolin-1在肝硬化门静脉高压症组脾静脉中较正常脾静脉中表达明显增强;eNOS在肝硬化门静脉高压症组脾静脉中呈低水平表达,较正常脾静脉中表达明显减少。结论肝硬化门静脉高压症组脾静脉中caveolin-1的过量表达及eNOS表达降低,导致NO合成减少,脾静脉血管阻力持续增加,及caveolin-1致静脉血管病理改变作用,共同作用致脾静脉出现对门静脉高压失代偿改变。     

ACTIVATION OF ENDOTHELIAL NITRIC OXIDE SYNTHASE IN CONTRALATERAL TESTIS DURING UNILATERAL TESTICULAR TORSION IN RATS

There are controversies about the injury of the contralateral testis during unilateral testicular torsion (UTT). An autonomic reflex arc between bilateral testes has been proposed. The authors focused on the involvement of nitric oxide (NO) in the contralateral testis during UTT. Eight-week-old male Wistar rats underwent unilateral torsion (1 h)-detorsion (up to 24 h). NO synthase (NOS) activity was detected as NADPH-diaphorase activity after fixation by paraformaldehyde. N -nitro- L -Arginine methyl ester (L-NAME, 20 mg/kg) was injected intravenously to the other group of rats. To evaluate the testicular injury, proteolysis of &#102 -fodrin production was detected by Western blotting. Apoptosis of the germ cells was evaluated by TUNEL. Long-term effect on spermatogenesis was evaluated by flow cytometry at 60 days after UTT. Transient activation of NOS was detected following the proteolysis of &#102 -fodrin in the contralateral testis. L-NAME inhibited these alterations. NADPH-diaphorase activity and eNOS immunoreactivity were co-localized in the endothelial cells. These reactions were not observed in other organs. There was neither enhanced apoptosis nor deteriorated spermatogenesis in the contralateral testis during and 60 days after UTT. In the contralateral testis, eNOS-derived NO regulates the vasomotor function against unilateral testicular torsion, whereas it acts slightly cytotoxic. These results suggest the possible involvement of a testis-specific neurovasomotor reflex between the bilateral testes.     

一氧化氮对体外保存血小板活化的影响研究

目的研究一氧化氮(NO)溶液对体外保存的手工分离血小板活化的影响。方法用高纯铜与稀硝酸反应,制备NO气体,然后溶解于蒸馏水中制备成NO饱和溶液。手工制备富含血小板血浆(PRP),在滤过白细胞前注入1/10PRP体积的不同浓度NO溶液,将血小板最大回收率时的NO溶液浓度定为实验的最适浓度,观察含此NO浓度的血小板保存5d内各项相关指标的变化,并进行统计学分析。结果NO饱和溶液10-2稀释时,血小板的回收率最大。加入NO组和对照组比较,血小板pH、MPV、PDW三项指标无统计学差异,但保存1天后CD62p再表达率有显著性差异(p<0.01)。结论NO可减少体外保存血小板的活化。     

睡眠剥夺复合贫铀污染对大鼠行为学及皮层NOS的影响

目的 观察睡眠剥夺（sleep deprivation,SD）复合贫（depleted uranium,Du）污删对大鼠的行为学及皮层一氧化氮合酶（NOS）的影响。方法 不同剂量贫铀颗粒气管灌注后3个月,采用小平台水环境法（flower pot）建立大鼠SD模型,观察大鼠SD 72 h后Y迷宫正确百分率及大脑皮层NOS数目变化情况。结果 DU 1mg＋SD组、DU 3mg＋SD组和DU5 mg＋SD组迷宫正确率比盐水对照（NS）＋SD组分别降低10．9％、14．1％和14．1％,差异有显著性（P〈0．05）,而与对应的单纯染铀组比较则分别降低17．4％、21．4％、17．9％,差异有显著性（P〈0．05）。Du 1mg组、DU 3mg组和DU 5mg组皮层NOS阳性神经元数目比NS组分别降低22．1％、19．7％、21．4％,差异有显著性（P〈0．05）;Du 1mg＋SD组、Du 3mg＋SD组和Du 5mg＋SD组比NS＋SD组分别降低26．0％、23．1％、27．9％,差异有显著性（P〈0．05）,而与对应的单纯染铀组比较则分别降低20．2％、19．6％、23．0％,差异有显著性（P〈0．05）。结论 SD能加重贫铀污染大鼠的学习记忆障碍,并使皮层nNOS阳性神经元数目明显减少,从而加重大鼠脑功能的损伤。     

龙血竭胶囊对早孕绒毛组织中一氧化氮合酶的影响

[目的]探讨龙血竭胶囊对早孕绒毛组织中一氧化氮合酶的影响。[方法]将自愿药物流产的早孕妇女随机分为两组：试验组加服龙血竭胶囊,对照组加服安慰剂。测定服药后绒毛组织中一氧化氮合酶（nitric oxide syn-thase NOS）的含量。[结果]绒毛组织中NOS的含量试验组显著低于对照组（P﹤0.05）。[结论]药物流产同时加服龙血竭胶囊能使绒毛组织中的NOS活性进一步降低,导致一氧化氮合成减少,从而减弱一氧化氮舒张血管和对血小板聚集的抑制作用,加强止血效果。     

阿托伐他汀对永久性大鼠局灶性脑缺血(pMCAO)的保护作用及其对脑eNOS表达的影响

[目的]观察阿托伐他汀（atrovastatin）对大鼠pMCAO（permanent occlusion of middle cerebral artery）模型脑eNOS（endothelial nitric oxide synthase）的表达及其表达规律的影响，探讨其具有脑保护作用的基础。[方法]采用线栓法制备pMCAO模型，并将试验动物随机分组：假手术组（A组），pMCAO组（B组），pMCAO＋阿托伐他汀组（C组），pMCAO＋提前给阿托伐他汀组（D组），每组再分为3个小组：2，3，7d组。比较各组nederson神经行为学评分、梗死体积，以及eNOS的表达。[结果]神经行为学评分，梗死后3d和7d，B、C两组之间差异有统计学意义（P〈0．05）；梗死体积测定，在2、3、7d，B组与C组、D组比较差异有统计学意义（P〈0．05）；C组与D组比较差异无统计学意义（P〉0.05）。eNOS阳性表达2、3、7d，各组之间比较差异有统计学意义（P〉0．05）。[结论]阿手巴伐他汀可提高大鼠的神经行为学评分，减少梗死体积，增加eNOS的表达，促使eNOS表达的增加是阿托伐他汀是有脑传护作用的基础。     

一氧化氮在病毒性肝炎病理生理过程中的作用

一氧化氮参与肝细胞多项生理功能的调节,参与病毒性肝炎的病理生理过程.在病毒性肝炎发病机理方面,一氧化氮可能具有保护肝脏和导致肝损害的双重作用.此外,一氧化氮还具有潜在的抗病毒活性及增加肝细胞癌变的危险性.本文综述了一氧化氮在病毒性肝炎病理生理过程中的这些作用.     

氯化锂对染铅大鼠海马nNOS蛋白与基因表达的影响

目的:探讨锂对醋酸铅神经毒性的拮抗效应及其可能机制。方法:通过ABC免疫组化法和半定量逆转录-聚合酶链式反应(RT-PCR)方法,研究饮用0.2g/L醋酸铅(PbAc)饮水和含不同剂量(3、30、300和3000mg/kg)氯化锂(LiCl)饲料喂养的大鼠海马nNOS蛋白与基因表达的变化。结果:与对照组比,3、30mg/kgLiCl组大鼠海马CA1区nNOS阳性神经元数目和nNOSmRNA相对含量显著增多;而染铅组大鼠则显著减少。Pb+LiCl(3、30和300mg/kg)组均较染铅组有显著性增加,但Pb+3000mg/kgLiCl组与染铅组无显著性差异。各组大鼠海马齿状回nNOS阳性神经元数目的变化与CA1区变化基本一致,CA3区变化不显著。结论:锂对铅的神经毒性有明显的拮抗作用。     

吸入一氧化氮疗法对犬吸入性损伤毒副作用的预防

为评价吸入一氧化氮（ＮＯ）对犬吸入性损伤毒副作用的影响和预防。将１７只犬随机分为对照和治疗组，均经烟雾吸入致伤后，对照组单纯吸Ｏ２；治疗组吸Ｏ２和ＮＯ混合气体。间断监测吸入混合气体中二氧化氨（ＮＯ２）浓度和动脉血变化。结果表明，ＮＯ２经钠石灰处理后比处理前降低８０．２％，治疗组动脉血高铁血红蛋白（ＭＨｂ）比对照组明显升高，在８和１２小时差异显著；动脉血浆亚硝酸盐（ＮＯ－２）和ＭＨｂ含量升高呈正相关。提示吸入ＮＯ疗法在混合气体中确实产生较高浓度ＮＯ２，动脉血ＭＨｂ浓度随吸入时间延长而逐渐升高     

一氧化氮对小鼠胚胎种植的影响

目的：评价一氧化氮对小鼠胚胎种植的影响。方法：受试昆明小鼠于妊娠第d3～d5分别腹腔注射NO供体硝普钠（SNP）1．0mg／kg、5．0mg／kg和10．0mg／kg，对照组小鼠注射等量的生理盐水；妊娠第6d颈椎脱臼法处死小鼠，肉眼观察每一侧子宫角孕囊数，形态计量学方法分析子宫内膜组织学变化，明胶酶谱法检测子宫内膜基质金属蛋白酶的表达。结果：①1．0mg／kgSNP组小鼠子宫角孕囊数与对照组比较无显著性差异（P〉0．05），5．0mg／kgSNP组小鼠宫角未观察到孕囊（P〈0．001），10．0mg/kg SNP组小鼠在注射药物20min后全部死亡；②1．0mg／kgSNP组与对照组小鼠子宫内膜均观察到明显的蜕膜样改变，丽组间腺体面积和间质面积比较无显著性差异（P〉0．05），5．0mg／kgSNP组小鼠子宫内膜未观察到蜕膜样改变，子宫内膜腺体面积明显低于对照组和1．0mg／kgSNP组（P〈0．001），间质面积明显高于对照组和1．0mg／kgSNP组（P〈0．001）；③1．0mg／kgSNP组与对照组小鼠子宫内膜均检测到酶原型MMP-2、激活型MMP-2和酶原型MMP-9的表达，5．0mg／kgSNP组小鼠子宫内膜仅检测到酶原型MMP-2的表达。结论：子宫内膜较高浓度的NO对胚胎种植具有明显的抑制效应，这种作用可能是通过影响MMP-2和MMP-9的表达及其活化以及干扰子宫内膜的蜕膜化实现的。     

二氧化氯灭活水中脊髓灰质炎病毒的机制

目的探讨二氧化氯对脊髓灰质炎病毒核酸和衣壳蛋白的损伤在病毒灭活中作用,阐明二氧化氯灭活脊髓灰质炎病毒机制。方法观察不同质量浓度（0．1、0．2、0．4、0．8、1．2mg／L）和不同作用时间（0、1、2,4、8和12min）二氧化氯对脊髓灰质炎病毒的作用,用大片段逐步步移RT-PCR（10ng-overlapping RT-PCR）分析二氧化氯对病毒全基因组的损伤,用ELISA分析病毒衣壳蛋白的损伤,通过细胞培养检测病毒感染性。结果脊髓灰质炎病毒衣壳蛋白对二氧化氯抵抗力强于核酸;基因组各部分对二氧化氯抵抗力强弱存在差异;二氧化氯对PV1 5’-NCR124-679nt区域的损伤与PV1的灭活一致。结论二氧化氯主要通过破坏脊髓灰质炎病毒核酸而不是衣壳蛋白来灭活病毒;病毒基因组5’-NCR内二级结构区域的破坏对脊髓灰质炎病毒具有致死效应。     

NITRIC OXIDE IS NOT INCREASED IN ALCOHOLIC BRAIN

Nitric oxide (NO) metabolites nitrite and nitrate were measuredin the cerebrospinal fluid in 12 alcohol-dependent subjectsand in 16 healthy controls. The amounts of NO metabolites inalcoholics were not different from those in the controls. Theresults suggest that NO is not a major factor responsible forbrain damage in these patients.     

DETECTION AND BIOIMAGING OF NITRIC OXIDE IN BOVINE OOCYTES AND SPERM CELLS

Mammalian gametes contain constitutive nitric oxide synthases (NOS) to synthesize nitric oxide (NO). The detection and bioimaging of NO in bovine gametes is important to determine the regulatory roles of NO during the different events of fertilization. Diaminofluoresceins, new fluorescence indicators for NO, were applied to detect the release of NO from bovine gametes. These compounds yield green fluorescent triazolofluoresceins, which provide the advantages of high specificity, sensitivity, and simplicity for the detection of NO. In this study, we mapped the expression of NOS in the bovine sperm and ova. NOS activity in sperm first appeared in the acrosome, then 60?min later in the head, middle piece, cytoplasmic droplet, and tail. Cow ova had high NO activity in the cytoplasm and in the surrounding corona cells, but not in the zona pellucida. These results show that for bovine gametes, the synthesis NO by the NOS system presents clear patterns of time and spatial distribution that may be important for the different events of fertilization.     

胃癌患者一氧化氮、超氧化物歧化酶及脂质过氧化物的变化

目的:研究胃癌患者一氧化氮、超氧化物歧化酶及脂质过氧化物的变化。方法:检测51 例胃癌患者( 患者组)和50 例健康成人( 对照组) 血浆一氧化氮(P- NO) 含量、红细胞超氧化物歧化酶(E- SOD) 活性、血浆过氧化脂质(P- LPO) 及红细胞过氧化脂质(E- LPO) 。结果:与对照组相比,患者组的P- NO、P- LPO 及E- LPO 平均值均显著升高( P< 0 .001) ,E- SOD 值显著降低( P< 0 .001) 。结论:提示胃癌患者体内的NO 代谢异常,脂质过氧化反应病理性加剧。     

EVIDENCE THAT CHANGES IN HIPPOCAMPAL EXCITABILITY IN VITRO ARE CAUSED BY WITHDRAWAL FROM CHRONIC IN VIVO ETHANOL ADMINISTRATION

A complex pattern of changes in the field potentials recordedfrom mouse hippocampal slices, prepared after chronic ethanoltreatment in vivo, has previously been demonstrated in thislaboratory. In the present study, recordings from slices preparedimmediately after 2 weeks of ethanol treatment, showed onlyan increase in paired pulse potentiation, compared with controls,whereas recordings made immediately after 16 weeks of ethanoladministration showed decreases in the thresholds for singleand multiple population spikes, increases in paired pulse potentiationand epileptiform activity. In hippocampal slices prepared after24 hr withdrawal, following 16 weeks of ethanol treatment, therewere no signs of hyperexcitability in the field potentials.Ratings of convulsive behaviour were increased in mice duringa 12-hr period after withdrawal from 16 weeks of ethanol treatment.Corresponding behaviour ratings for the mice given ethanol for2 weeks, or those withdrawn for 24 hr after 16 weeks ethanoltreatment, were not significantly different from control values.It was concluded that epileptiform activity seen in hippocampalslices after prolonged ethanol administration may contributeto the ethanol withdrawal hyperexcitability seen in vivo.