Nitric Oxide:from a mysterious labile factor to the molecule of the Nobel Prize Recent progress in nitric oxide research

NO is now known to be an important messenger molecule in biology.It regulates a variety of functions within cells and tissues including vasodilation, neurotransmission and immunological process. This review will focus on the nitric oxide synthase gene family and recent progress on molecular genetic analysis of NOS1, NOS2 and N0S3 genes.

Extract of Meretrix meretrix Linnaeus induces angiogenesis in vitro and activates endothelial nitric oxide synthase

Meretrix meretrix Linnaeus has long been used as traditional Chinese medicine in oriental medicine.The angiogentic activity of the extract of M.meretrix was investigated in this study,using human umbilical vein endothelial cells(HUVECs).Extract of M.meretrix Linnaeus(AFG-25) was prepared with acetone and ethanol precipitation,and further separated by Sephadex G-25 column.The results show that AFG-25 promoted proliferation,migration,and capillary-like tube formation in HUVECs,and in the presence of eNOS inhibitor NMA,the tube formation induced by AFG-25 is inhibited significantly.Moreover,AFG25 could also promote the activation of endothelial nitric oxide synthase(eNOS) and the resultant elevation of nitric oxide(NO) production.The results suggested that M.meretrix contains active ingredients with angiogentic activity and eNOS/NO signal pathway is in part involved in the proangiogenesis effect induced by AFG-25.

Studies on the Compounds of d4T Combined with Nitric Oxide Donors and Nitric Oxide Synthase Inhibitors and their Anti-HIV and AIDS Activity

Stavudine, a potent anti HIV and AIDS related complex, is one of the Nucleoside Analogue Reverse Transcriptase Inhibitors (NARTIs). It is phosphorylated intracellularly and then inhibits the viral reverse transcriptase by acting as a false substrate. Modifications made on the hydrogen labile at the 5' position on the sugar is an interesting template for the elaboration of new potent anti HIV and AIDS drugs. The expected advantages of the modified stavudine prodrugs can be multiple: synergistic drug activities, enhancement of stavudine intracellular uptake, increase of stavudine brain delivery, and bypass of the first stavudine phosphorylation step into the cells. Nitric oxide synthase inhibitors of stavudine and nitric oxide donors of stavudine may hold significant promise for the treatment of HIV and AIDS.

Effects of Infrasound on Gastric Motility, Gastric Morphology and Expression of Nitric Oxide Synthase in Rat

Infrasound widely condition, productive exists in nature, our living and traffic environment. Gastrointestinal tract is relatively sensitive to infrasound. However, the effect of infrasound on gastrointestinal function is unclear. Therefore, the purpose of our study was to observe the effects of infrasound on gastric motiliW and gastric morphology and to assess the expression of nitric oxide synthase （NOS） in gastric antrum after exposure to infrasound of 8 Hz - 130 dB for 2 hours per day for 14 consecutive days. Gastric motility was assessed by gastric fluid-emptying rate. Gastric morphology was evaluated by HE. The expression of NOS was measured by tissue microarray technology. The results would contribute to understand the role of infrasound in gastroenterology, and help to explain the mechanism of infrasound on gastroenterology.

Effects of Siqi Decoction on Concentration of Nitric Oxide and Activity of Nitric Oxide Synthase in Blood Serum of Rats with Myocardial Ischemia

To study the effects of Siqi decoction on rats with Myocardial Ischemia, the model of Myocardium Ischemia was made for the Wistar rats cured with posterior pituitary injection through vein in tail. Siqi decoction, Diaoxinxuekang（DK） and Fufangdanshenpian（FD）, the latter two drugs of which were effective TCM drugs of anti-Myocardial Ischemia at present, were administrated to the rats with Myocardium Ischemia for 5 days to compare the effect of anti-Myocardium Ischemia as reference drugs by measuring the changes of NO concentration and activity of NOS in the rat blood serum with Myocardial Ischemia. There was a remarkable increase in the NO concentration and activity of NOS in serum in Siqi decoction groups compared with those in control group（p〈0.05）. The results of the prevention group in experiment of Siqi decoction are better than those of the cure group. Siqi decoction was really fit for Myocardium Ischemia via increasing NO concentration by stimulating the activity of NOS in serum. The effect of Siqi decoction against Myocardium Ischemia in preventive group is better than the curative that of Siqi decoction in the curative group.

N^ω-Nitro-N^ω’-Substituted Guanidines: A Simple Class of Nitric Oxide Synthase Inhibitors

A series of Nω-nitro-Nω’-substituted guanidines has been prepared as potential inhibitors of the human Nitric Oxide Synthase (NOS) isoforms. The reported utility of amino-guanidine and nitroarginine in iNOS inhibition points to a potential similar utility for analogs of nitro-guanidine. The compound library was tested against the three isoforms of Nitric Oxide Synthase (eNOS, iNOS and nNOS). Several candidates showed excellent activity and good selectivity for nNOS. One particular compound even demonstrated good selectivity for iNOS. The potential usefulness of such selective inhibitors is discussed.

Expression of nitric oxide synthase in the spinal cord after selective brachial plexus injury

BACKGROUND: Some researches showed that motoneurons in spinal cord anterior horn wound die following brachial plexus injury, but the concrete mechanism of motoneurons death remains unclear. OBJECTIVE: To observe the expression of nitric oxide synthase (NOS) and survival of C7 motoneurons in spinal cord of rats after selective brachial plexus injury. DESIGN: A randomized controlled animal experiment. SETTING:Department of Anatomy, Sun Yet-sen Medical College, Sun Yet-sen University. MATERIALS: Totally 35 adult healthy male Sprague-Dawley rats with the body mass of 200-300 g were provided by Experimental Animal Center, Sun Yet-sen Medical College, Sun Yat-sen University. The rats were divided into control group (n =5) and experimental group(n =30) by random number table method, and the experimental group was divided into three injury subgroups: anterior root avulsion group, dorsal root transection group and spinal cord hemisection group, 10 rats in each group. There were horse anti-neuronal NOS (nNOS) polycolonal antibody (Sigma company) and nicotina mideadeninedinucleotide phosphate (NADPH-d) (Sigma Company). METHODS: The experiment was performed at Department of Anatomy, Sun Yet-sen Medical College, Sun Yet-sen University between September 2004 and April 2005. ①After anesthetizing the rats, the spinous process of second thoracic vertebra as a marker, the vertebra was exposed from C5 to T1 and the lamina of vertebra was unclenched, and spinal dura mater was carved to expose the spinal nerve dorsal roots of C5-T1. The right ventral root of C7 was avulsed, and the residual root was removed in anterior root avulsion group. The right ventral root of C7 was avulsed and the right dorsal roots of brachial plexus (C5-T1) were cut off in dorsal root transection group. In spinal cord hemisection group, the hemisection between the C5 and C6 spinal segment on right side and avulsion of right ventral root of C7 were made. In the control group, the vertebra from C5 to T1 was unclenched and the skin of wound was sutured. ②Three weeks after operation, behavior of rats was observed. The rats were killed after anesthesia. The C7 segment of spinal cord was removed and treated with NADPH-d staining, neutral red counterstaining and NOS immunohistochemistry staining to detect the expression of NOS. MAIN OUTCOME MEASURES: The expression of NOS and survival of C7 motoneurons in spinal cord of rats 3 weeks after operation. RESULTS: Among the 35 included rats, 3 rats died 2 weeks following operation, so totally 32 rats were involved in the result analysis. ①NADPH-d positive neurons of in anterior horn of C7 in the three groups: The NADPH-d positive neurons could be found in anterior horn of C7 in the three groups. The percentage of that in anterior root avulsion group to that of non-injury side of spinal cord was(20.98±2.65)%, (29.43±6.81)% in dorsal root transection group and (31.74±6.80)% in spinal cord hemisection group. There was significant difference among the three injury groups(F =5.135,P =0.016). There was significant difference in anterior root avulsion group with dorsal root transection group and spinal cord hemisection group (t =2.562,3.167,P < 0.05). There was no significant difference between the dorsal root transection group and spinal cord hemisection group (P =0.534). ②survival rate of motoneurons in anterior horn of C7: There were dead motoneurons in the three injury groups, the percentages of surviving motoneurons to that of non-injured side of spinal cord were (69.22±4.04)%,(62.01±3.82)% and (56.74±6.86)%, respectively. There were significant differences among the three groups (F =9.508,P =0.002). The anterior root avulsion group was significantly different from the other two groups(t =2.764,4.587,P < 0.05). There was no significant difference between the dorsal root transection group and spinal cord hemisection group(P =0.073). CONCLUSION: The selective brachial plexus injury can induce the up-regulation of NOS expression in motorneurons of spinal cord anterior horn and block descending pathway of cortex to cause the more significant up-regulation of NOS and low survival rate in motoneurons. It indicates that descending pathway of cortex can inhibit the NOS expression in motorneurons of spinal cord anterior horn, and the high NOS expression might induce the death of motorneurons in spinal cord anterior horn.

Coexistence of nitric oxide synthase with γ-aminobutyric acid during seizure induced by kainic acid in SD rats

Objective: Functional significance of NO and central inhibitory neurotransmitter γ-aminobutyric(GABA) during seizures were investigated morphorlogically. Methods: A kainate-induced complex partialseizure model was used in our experiment. Twenty SD rats were randomly divided into KA 30, 60, 90, 200min and control groups. The brain sections were stained by NADPh (nicotinamide adenine dinucleotide phosphate ) diaphorase (Nd ) histochemically, and were further stained by GABA immunohistochemically.Results: Histological and immunohistochemical study revealed that in KA groups the number of Nd and GABA-positive double labelled neurons in CA3 region, CA3 region and dentate gyms was significantly reduced,compared with the control group. Conclusion: Nd coexisted with GABA in the brain. Reduction of GABA release led to relief of GABA-ergic inhibition and in the same way, reduction of NO release weakened its negative feedback modulation. Therefore neuronal synchronous paroxysmal discharges increased. GABA and NO,both having antiepileptic action, acted through different ways or different link in the same way. NO may involve in the effect of GABA-ergic neurons and play cooperative antiepileptic action with GABA.

大骨节病患者血清NO、NOS、sFas/APO-1检测分析

目的探讨一氧化氮(NO)、一氧化氮合酶(NOS)、诱导型一氧化氮合酶(iNOS)、可溶性Fas(sFas)诱导大骨节病发生发展的作用与特点,为大骨节病防治研究提供新的思路。方法选择陕西省永寿县、榆林市榆阳区大骨节病区与咸阳市渭城区非大骨节病区120例调查对象,分为成人、儿童大骨节病组;成人、儿童病区对照组;成人、儿童非病区对照组,每组20例,平均年龄、性别无差别。用酶还原法检测了血清NO,化学比色法检测了血清NOS、iNOS,ELISA法检测了血清sFas水平。结果①大骨节病成人组血清NO、iNOS显著高于病区和非病区成人对照组(P< 0.05);血清NOS、sFas/APO鄄1水平高于非病区成人对照组(P< 0.005)。②大骨节病儿童组血清NO高于非病区儿童对照组(P< 0.05);血清NOS、iNOS均高于病区和非病区儿童对照组(P< 0.005)。结论儿童大骨节病发生发展中NO诱导途径起一定的作用,而Fas途径作用不明显。当疾病发展到成人大骨节病时,二者均起一定作用。

氯胺酮对大鼠不同脑区NOS活性和NO产量的影响

目的：了解氯胺酮对大鼠脑ＮＯＳ活性和ＮＯ产量的影响。方法：１６只ＳＤ大鼠，随机分为二组，分别ｉｐ生理盐水１０ｍｌ／ｋｇ（对照组）和氯胺酮１００ｍｇ／ｋｇ。用分光光度法测定ＮＯＳ活性和ＮＯ产量。结果：大鼠ｉｐ氯胺酮１００ｍｇ／ｋｇ能明显抑制小脑、大脑皮层、海马的ＮＯＳ活性和减少ＮＯ的产量（Ｐ＜００５或Ｐ＜００１），脑干的ＮＯ产量亦较对照组减少（Ｐ＜００１），ＮＯＳ活性下降，但无统计学意义。结论：大鼠ｉｐ氯胺酮１００ｍｇ／ｋｇ能明显抑制脑ＮＯＳ活性和减少ＮＯ产量。

从NO、NOS变化探讨瓜蒌薤白白酒汤对心肌缺血再灌注损伤的防治作用

目的:研究瓜蒌薤白白酒汤(LGXD)对家兔心肌缺血再灌注损伤模型血清中一氧化氮(NO)含量和一氧化氮合酶(NOS)活性的影响,并探讨其对再灌注损伤的可能性保护机制。方法:采用结扎左冠状动脉前降支法复制心肌缺血再灌注损伤模型,分离血清检测血清中一氧化氮(NO)含量和一氧化氮合酶(NOS)活性,摘取左室心肌做常规病理切片。结果:与假手术组比较,模型组NO含量和NOS活性均显著增高,与模型组比较,高、低剂量药物组NO含量和NOS活性均降低,低剂量组比高剂量组低,病理结果显示低剂量组优于高剂量组。结论:瓜蒌薤白白酒汤对家兔心肌缺血再灌注损伤的心肌有保护作用,其机制可能与抑制NOS的活性,减少NO的过量产生有关,且低剂量药物组疗效优于高剂量组。

2型糖尿病合并高血压患者血清NO、NOS水平的研究

目的 通过对 2型糖尿病 (2 - DM)合并高血压 (EH)患者血清一氧化氮 (NO)、一氧化氮合酶 (NOS)的测定 ,探讨其在 2 - DM合并 EH发生发展中的意义。方法 设正常对照组、单纯 2 - DM组和 2 - DM合并 EH组 ,分别测定血糖、胰岛素、血脂、NO及 NOS。结果 2 - DM合并 EH组空腹胰岛素 (FINS)、舒张压 (DBP)和收缩压 (SBP)等均显著高于单纯 2 - DM组和正常对照组 ;胰岛素敏感指数 (ISI)和 NO均显著低于其他两组。相关分析显示 NO与 FINS和血压呈明显负相关 ,与 ISI呈显著正相关。结论 血清中 NO含量与胰岛素抵抗和血压密切相关 ,2 - DM患者血清 NO水平的降低可能参与了 2 - DM患者合并 EH的发病。

脑缺血后脑组织NO含量和NOS活性变化及三七总皂甙的影响

为了探讨一氧化氮 (NO)在缺血神经元坏死中的作用机制及三七总皂甙 (PNS)是否通过影响NO的变化而起脑保护作用。 方法 用栓线法建立大鼠局灶性脑缺血模型 ,测定脑缺血后不同时间脑组织NO含量和一氧化氮合成酶 (NOS)活性变化及PNS对其的影响。 结果 缺血后 3 0分钟脑组织NO含量和NOS活性均显著升高 (P <0 0 1 ) ,而PNS能防止脑缺血后两者的升高。NO含量与NOS活性呈显著正相关。 结论 NO在缺血神经元损伤中起重要作用 。

荆花胃康胶丸对溃疡大鼠胃黏膜NO,NOS和ET含量的影响

目的:观察荆花胃康胶丸对溃疡大鼠胃黏膜内皮素(ET)、一氧化氮(NO)、一氧化氮合酶(NOS)含量的影响。方法:采用Okabe氏法造成大鼠胃溃疡模型,随机将模型动物分为蒸馏水对照组、荆花胃康胶丸30,20, 10 mg·kg^(-1)·d^(-1)组、硫糖铝10 mg·kg^(-1)·d^(-1)及法莫替丁5 mg·kg^(-1)·d^(-1)组,每组8只。各组均灌胃给药,qd,连续10 d。末次给药后次日,测定各组溃疡面积,并测定溃疡周围组织NO,NOS及ET含量。结果:与对照组相比,荆花胃康胶丸30,20,10 mg·kg^(-1)·d^(-1)组的溃疡面积显著降低,溃疡周围组织NO及NOS的含量明显增高,ET的含量明显降低(P＜0．01),且呈现剂量依赖性;荆花胃康胶丸30 mg·kg^(-1)·d^(-1)组的保护作用优于硫糖铝及法莫替丁组(P＜0．05)。结论:荆花胃康胶丸可能通过增加胃溃疡组织NO及NOS的含量及降低ET的含量来发挥胃黏膜修复作用。

四种复方中药和黄芪多糖对鲫鱼生长、组织中NO含量与NOS活性的影响

将四种复方中药(分别编号为AB、AH、CD、IV)和黄芪多糖(APS)作为添加剂添加入基础料饲料中,连续饲喂1龄鲫鱼45d,于第45d及第60d分别采样,测定鱼体重和肝脏、脾脏、肾脏中一氧化氮(NO)含量及一氧化氮合酶(NOS)活性,研究中药复方对鲫鱼生长及免疫功能的影响。结果显示:45d时称量体重,复方AH、复方AB两组增重率显著高于对照组及其他三种中药复方组,饲喂60d对照组及所有中药组增重率均无显著差距。投喂45d,方剂AB显著降低鲫鱼肝脏、肾脏中NO的含量,五种中药方剂均能提高鲫鱼脾脏中NO含量。AH、AB、IV、APS四种方剂饲喂的鲫鱼肝脏中NOS含量明显高于对照组。投喂60天,五种中药方剂饲喂的鲫鱼脾脏中NO含量均高于对照组,除IV外,均差异显著。肝脏中NOS含量各组无显著差异,脾脏中IV组TNOS含量最高,APS组iNOS含量最高。肾脏中APS方剂组的iNOS含量显著高于对照组。

注射L-精氨酸和环磷酰胺对杂色鲍血清NO水平、NOS活性及免疫指标的影响

对杂色鲍(Haliotis diversicolor)分别按每kg体质量注射250mg、500mg和1000mg的L-精氨酸和5mg、10mg、20mg的环磷酰胺。每隔5d足部肌肉注射1次,共注射3次。第16天检测其血清中NO含量以及NOS活性的变化情况。同时采用注射环磷酰胺的方法(剂量为10mg/kg体质量),对杂色鲍血清中NOS活性进行负调控,分别在注射后3h、6h、12h、24h、48h、96h对血清NO水平和血清NOS、超氧化物歧化酶(SOD)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)以及溶菌酶(LZM)活性进行测定,并探讨它们之间的相关性。结果显示,注射500mg/kg的L-精氨酸可以显著提高实验鲍血清中NO水平和NOS活性,而注射10mg/kg的环磷酰胺则可以显著降低实验鲍血清中NO水平和NOS活性。血清中NOS活性与ACP、AKP以及LZM等活性的相关系数分别为0.8074、0.8292和0.7408,相关显著(P<0.05)或极显著(P<0.01),NO与SOD及LZM的相关系数分别为0.9302和0.9413,相关极显著(P<0.01)。实验结果为人工调控鲍体内的NO含量、增强其自身的非特异性免疫功能,从而提高鲍的抵抗疾病的能力提供了科学依据,同时也证明NO和NOS可以作为评价鲍免疫功能强弱的指标。

NO、NOS与宫缩乏力性产后出血关系的研究

目的:探讨血清NO及NOS在预测宫缩乏力性产后出血发生发展中的作用。方法:选择剖宫产终止妊娠的孕妇180例,因臀位、社会因素、骨盆狭窄、胎儿宫内窘迫等行剖宫产术,按产后24 h内出血量≥500 m l分为产后出血组(n=47)及非产后出血组(n=133)。所有孕妇于剖宫产术麻醉前30 m in及术后2 h测血清NO及NOS,术中娩出胎儿后于子宫切缘上缘及胎盘母体面中央无钙化区分别取子宫肌组织及胎盘组织各一,切片作免疫组织化学分析iNOS的表达,准确收集剖宫产术中出血量及产后24 h出血量。结果:产后出血组术前、术后血清NO及NOS含量较非产后出血组明显增高,差异有显著性意义(P<0.05);血清NOS预测产后出血的灵敏度、特异度、阳性预测值、阴性预测值均高于血清NO值;从SABC免疫组化结果可见在子宫肌及胎盘均有iNOS表达,产后出血组子宫肌和胎盘的iNOS阳性表达率均显著高于非产后出血组,P<0.05;经等级相关检验,子宫肌、胎盘组织iNOS的表达强度随术前血清NO、NOS水平的升高而增加,均呈正相关。结论:血清NO、NOS水平可以正确反映子宫、胎盘NOS的活性,可作为预测宫缩乏力性产后出血的1项有效的指标。

慢性低氧及运动训练对大鼠血清一氧化氮(NO)含量及一氧化氮合酶(NOS)活力的影响

为观察慢性低氧及训练对大鼠的影响,取健康SD大鼠28只,随机分成4组:(1)常氧对照组(NC),(2)常氧训练组(NT),(3)低氧安静组(HC),(4)低氧训练组(HT)。其中HC、HE两个低氧组每天保证22h生活在模拟4000m高原的低氧舱(氧浓度12 7%),NT、HT两个训练组则每天进行跑台训练1h。28天后,各组均于安静状态下宰杀、取血,测定血清一氧化氮(NO)含量及一氧化氮合酶(NOS)活力。结果可见:28天后,与对照组相比,低氧安静组(HC)和低氧训练组(HT)大鼠血清NO浓度有较显著的降低(P<0 10),而常氧训练组(NT)下降不显著;另一方面,慢性28天后,与常氧对照组(NC)相比,常氧训练组(NT)及低氧对照组(HC)NOS活性虽有上升,但差异不显著;低氧训练组(HT)NOS活性则明显低于NT组及HC组(P<0 05)(P<0 10),这说明低氧及运动两种因素的效应并非是简单的叠加,可能相互抵消。其机制有待进一步研究。

枸杞多糖对衰老小鼠脑NO、NOS的影响

目的 :研究枸杞多糖对衰老小鼠NO、NOS的影响。方法 :采用老年小鼠和D 半乳糖致衰老小鼠 ,分别连续灌服枸杞多糖 2 0mg/kg·d两个月和一个月 ,应用Griess法检测脑NO含量 ,双波长分光光度计测定NOS活性。 结果 :老年小鼠和D 半乳糖致衰老小鼠脑NO含量、NOS活性明显低于青年小鼠和正常对照小鼠 ,给药后则可明显提高之。结论 :枸杞多糖能明显提高衰老小鼠脑NO含量和NOS活性。

耐力训练与补充不同剂量L-Arg对大鼠股外肌NOS基因表达的影响

目的 :本研究旨在探讨耐力训练和补充不同剂量外源性L -Arg对耐力训练后和力竭运动恢复期大鼠股外肌NOS基因表达的影响。方法 :采用定量反转录聚合酶链式反应 (QRT -PCR)检测股外肌NOS三种亚型 (eNOS、nNOS、iNOS)的基因表达。结果 :耐力训练显著上调eNOS、nNOS表达 (P <0 .0 1) ,有上调iNOS表达的趋势 ;补充小剂量L -Arg可下调nNOS的表达 (P <0 .0 5 ) ,有上调iNOS表达的趋势 ;补充大剂量上调nNOS表达 (P <0 .0 5 ) ,有下调iNOS表达的趋势。结论 :耐力训练上调eNOS、nNOS、iNOS的表达 ;补充外源性L -Arg对nNOS。