血脂康对急性冠脉综合征患者内皮功能的影响

目的观察血脂康对急性冠脉综合征患者血清一氧化氮（NO）及一氧化氮合酶（NOS）的影响。方法随机选择急性冠脉综合征病患者67例．将其随机分为血脂康治疗组35例，阿托伐他汀组32例，并对两组患者治疗前后血清NO及NOS含量水平进行对比分析。结果血脂康和阿托伐他汀均可升高急性冠脉综合征患者血清NO及NOS水平，其作用无明显差异（P〉0．05）。结论血脂康可抑制脂质过氧化反应．保护血管内皮功能，对急性冠脉综合征的防治具有重要意义。     

一氧化氮合成酶抑制剂的研究进展

一氧化氮（NO）具有广泛的生理功能。哺乳动物体内的NO是由NO合成酶（NOS）氧化L－精氨酸而合成的,合成后的NO迅速跨膜扩散释放。NO合成失调能介导多种疾病产生。特异性NO抑制剂能通过调控NO的合成,对NOS表达相关的各种疾病的防治具有重要的临床意义。本文对近年来NOS抑制剂的研究作一概述。     

一氧化氮合酶抑制剂的研究进展

一氧化氮（NO）具有广泛的生理功能,哺乳动物体内的NO是由NO合酶（NOS）氧化L－精氨酸而合成的,合成后的NO迅速跨膜扩散释放,NO合成失调能介导多种疾病产生,特异性NOS抑制剂能通过调控NO的合成,对NOS表达相关的各种疾病的防治具有重要的临床意义,本文对近年来NOS抑制剂的研究作一概述。     

Cannabinoid Regulation of Nitric Oxide Synthase I (nNOS) in Neuronal Cells

In our previous studies, CB₁ cannabinoid receptor agonists stimulated production of cyclic GMP and translocation of nitric oxide (NO)-sensitive guanylyl cyclase in neuronal cells (Jones et al., Neuropharmacology 54:23–30, 2008). The purpose of these studies was to elucidate the signal transduction of cannabinoid-mediated neuronal nitric oxide synthase (nNOS) activation in neuronal cells. Cannabinoid agonists CP55940 (2-[(1S,2R,5S)-5-hydroxy-2-(3-hydroxypropyl) cyclohexyl]-5-(2-methyloctan-2-yl)phenol), WIN55212-2 (R(+)-[2,3-dihydro-5-methyl-3-[(morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazinyl]-(1-naphthalenyl)methanone mesylate), and the metabolically stable analog of anandamide, (R)-(+)-methanandamide stimulated NO production in N18TG2 cells over a 20-min period. Rimonabant (N-(piperidin-lyl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-H-pyrazole-3-carboxamide), a CB₁ receptor antagonist, partially or completely curtailed cannabinoid-mediated NO production. Inhibition of NOS activity (N ^G -nitro-l-arginine) or signaling via Gi/o protein (pertussis toxin) significantly limited NO production by cannabinoid agonists. Ca²⁺ mobilization was not detected in N18TG2 cells after cannabinoid treatment using Fluo-4 AM fluorescence. Cannabinoid-mediated NO production was attributed to nNOS activation since endothelial NOS and inducible NOS protein and mRNA were not detected in N18TG2 cells. Bands of 160 and 155 kDa were detected on Western blot analysis of cytosolic and membrane fractions of N18TG2 cells, using a nNOS antibody. Chronic treatment of N18TG2 cells with cannabinoid agonists downregulated nNOS protein and mRNA as detected using Western blot analysis and real-time polymerase chain reaction, respectively. Cannabinoid agonists stimulated NO production via signaling through CB₁ receptors, leading to activation of Gi/o protein and enhanced nNOS activity. The findings of these studies provide information related to cannabinoid-mediated NO signal transduction in neuronal cells, which has important implications in the ongoing elucidation of the endocannabinoid system in the nervous system. Howlett and Norford are guarantors of this work.     

大鼠生后肾内皮型一氧化氮合酶的定量分析

为探讨内皮型一氧化氮合酶在大鼠生后肾发育中的作用,采用SD大鼠按生后年龄随机分为6组,即新生组、生后3天组、5天组、7天组、14天组和成年组,用免疫印记技术和光密度分析法对各组大鼠生后肾内皮型一氧化氮合酶进行定量检测,以测得的一氧化氮合酶最大量为1(100%),计算蛋白相对含量,SPSS10.0统计软件包统计分析。结果显示,新生组酶含量最高为1,随年龄增长酶含量逐渐减少,至第7天达到最少为44.60±2.41%,以后又增高至14天达成年水平为71.55±4.35%。提示大鼠生后不同年龄组肾内皮型一氧化氮合酶有明显的变化规律,NO可能在肾脏的成熟与发育中起重要作用。     

内皮型一氧化氮合酶基因多态性与老年高血压关系的研究

目的:探讨内皮型一氧化氮合酶(eNOS)基因894G/T多态性与老年人原发性高血压病(EH)之间的关系。方法:应用聚合酶链反应限制性片段长度多态性技术对湖北地区汉族129例高血压病患者(EH组)和117例健康者(NT组)的eNOS外显子894位点进行基因分型,生化技术测定其血脂水平。结果:EH组T等位基因频率以及GT+TT基因型分布频率均显著高于NT组(P<0.05);在年龄不同组中这种显著相关性仅存在年龄小于61岁的中青年患者中。结论:eNOS基因可能与老年人群原发性高血压发生无关。     

Complexes of an Alpha Thymosin Derivative with (185/187) Re and (99m) Tc: Structural Analysis and Initial Biological Evaluation

Alpha thymosins are a family of immunostimulating peptides first isolated from thymus gland. In the present work, the structure of the ReO(V)(3+) complex of an alpha 1 thymosin derivative containing the metal-chelating N,N-dimethylglycyl-l-seryl-l-cysteinyl group was studied with NMR, CD, and ESI. The analysis indicated the existence of two interconverting diastereomers depending on the orientation of the side chain of the chelated Ser syn- or anti- to the oxygen of the ReO(V)(3+) core. The two diastereomers could be separated on HPLC under a slow gradient showing the ratio of syn/anti to be 3:2, in agreement with the NMR data. The conversion process was shown to involve the coordination of a water molecule to the ReO(V)(3+) core through the incubation of the complex in (18) O-enriched water and subsequent ESI analysis. HPLC analysis of the analogous radioactive (99m) TcO(V)(3+) complex showed the formation of two isomers in the same syn/anti 3:2 ratio. Biodistribution studies of the (99m) TcO(V)(3+) complex in Swiss albino mice with experimentally induced inflammation showed higher accumulated radioactivity in inflamed tissue compared to normal (ratio of inflamed/control tissue 3.9). (99m) Tc-labeled complexes of alpha thymosin derivatives are expected to facilitate research on alpha thymosins and accelerate exploitation of these peptides in immunotherapy protocols.     

Pharmacokinetics and Pharmacodynamics of 7-Nitroindazole,a Selective Nitric Oxide Synthase Inhibitor,in the Rat Hippocampus

Purpose. This study was conducted to assess the pharmacokinetics and pharmacodynamics of 7-nitroindazole (7-NI), a selective inhibitor of neuronal nitric oxide synthase (NOS). Methods. Male Sprague-Dawley rats were equipped with peritoneal/venous cannulae and a microdialysis probe in the hippocampal cortex. Rats received 7-NI in peanut oil (25 mg/kg) ip every 2 h for 14 h or peanut oil alone. Blood samples were obtained at timed intervals for serum 7-NI; brain tissue microdialysate for determination of extracellular 7-NI and NO was obtained every 20 min. A pharmacokinetic-pharmacodynamic model was constructed to evaluate the effects of 7-NI on NOS activity. Results. Consistent with previous reports, NOS activity in controls evidenced circadian variation. These cyclic changes in NO production were incorporated into the model of 7-NI effects on NOS. 7-NI produced a rapid (within 2 h) decrease in hippocampal NO. Under the conditions of this experiment, 7-NI produced an 50% decrease in hippocampal NO, which was sustained during 7-NI administration. The decrease in NOS activity by 7-NI was concentration-dependent with an apparent IC₅₀ of 17 g/ml. Conclusions. Multiple ip injections of 7-NI result in a predictable, sustained decrease in NO production in the hippocampus. The pharmacokinetic-pharmacodynamic model developed allows design of dosing regimens that can produce designated changes in brain NO content, facilitating use of 7-NI to probe the pharmacological implications of NO in the central nervous system.     

豚鼠耳蜗核复合体及脑干内一氧化氮合酶阳性神经元的分布

目的 :研究一氧化氮合酶 (NOS)在耳蜗核复合体 (CNC)及脑干的分布特点 ,探讨一氧化氮 (NO)在听觉径路中的作用 ;方法 :采用依赖还原型辅酶Ⅱ的尼克酰胺腺嘌呤二核苷酸磷酸黄递酶 (Nicotinamideadeninedinucleotidephosphate -Diaphorase ,NADPH -d)组织化学方法 ,研究了豚鼠听觉核团和脑干内NOS的分布 ;结果 :在脑桥的各级听觉传入核团 ,均有NOS阳性神经元 ,而上橄榄复合体NOS反应阴性 ;耳蜗核有NOS阳性神经元 ,其NOS阳性神经元主要集中在后腹核 ,为圆形或椭圆型双极细胞 ;下丘的臂内侧核NOS阳性神经元呈小的、圆形细胞。在脑干中 ,NOS阳性神经元分布广泛 ;在三叉神经中脑核 (Me5 )、蓝斑 (LC)、前庭内侧核(MVe)、MVe复侧核 (MVeV)、前庭外侧核 (Lve)、舌下神经前置核 (PrH)、巨细胞网状核 (Gi)、Gi腹侧部 (GiV)、外侧巨细胞旁核 (LPGi)、背侧巨细胞旁核 (DPGi)均有NOS阳性神经元 ;结论 :NO可能是听觉中枢的神经递质或调质 ,参与声信号传递的调节 ;并可能与脑干功能的调节有关。     

弥漫性颅脑损伤对大鼠海马一氧化氮合成酶活性及表达的影响

目的 :研究大鼠弥漫性颅脑损伤后海马结构一氧化氮合成酶 (NOS)活力及表达 ,分析海马不同亚区NOS活力变化与时间的关系。方法 :制作Wistar大鼠弥漫性脑损伤模型 ,并分对照组、假手术组及伤后6、12、24、48h组 ,于不同时间点获取脑组织 ;用NADPH 黄递酶 (NADPH d)组织化学法和免疫组化法检测NOS的活性及表达情况。结果 :NADPH d组织化学法显示 ,弥漫性脑损伤后 ,海马结构NOS阳性神经元数量在伤后6h最多 ,以后逐渐减少 ,伤后24、48h组明显低于假手术组 (P<0 05) ;免疫组化结果显示 ,在CA1、CA3区和齿状回 (DG) ,伤后24、48h组阳性细胞数均低于假手术组 (P<0 01)。结论 :大鼠弥漫性脑损伤后 ,可引起海马各区NOS的变化 ,该变化可能是造成脑组织继发性损害机制之一。     

eNOs基因T786C多态性与原发性高血压的关系

目的:探讨内皮型一氧化氮合酶(eNOs)基因T786C多态性与原发性高血压的关系。方法:选取原发性高血压患者266例(高血压组)和健康人群100例(对照组),入院后空腹12h抽取肘正中静脉血4mL,常规测定生化及凝血功能等各项指标。进行eNOs基因T786C的PCR扩增、酶切和基因型分析。分析不同基因型与高血压发病的关系。结果:高血压组的年龄、收缩压、舒张压、体质量指数及有饮酒史者均高于对照组,差异均有统计学意义(P<0.05或P<0.01);其他指标比较差异无统计学意义(P>0.05)。2组TT、TC及CC基因型分布和T、C等位基因频率分布差异均无统计学意义(P>0.05)。高血压组患者中C等位基因携带者的收缩压及舒张压与TT基因型者差异无统计学意义(P>0.05)。影响高血压的因素有年龄、性别和体质量指数。结论:eNOs基因T786C多态性在高血压发病中不起直接作用,可能通过影响相关危险因素参与高血压发病。     

花生四烯乙醇胺对大鼠心功能与心肌一氧化氮水平及一氧化氮合酶活性的影响

目的 观察大麻素类物质花生四烯乙醇胺（anandamide，Ana）对大鼠体外心脏左心室心肌中一氧化氮(NO)含量和一氧化氮合酶(NOS)活性的影响。方法 采用Langendorff方法观察Ana对大鼠体外心脏心率（HR）、冠脉流量（CF）、冠脉灌注压(CPP)、左室压最大上升速率(＋dp/dtmax)、左室压最大下降速率( dp/dtmax)、左室收缩峰压(LVSP)、左室舒张末压(LVEDP)及左室发展压(LVDP)的影响；NO含量和NOS活性采用联苯胺荧光分光光度法测定。结果 Ana可使体外心脏心率、CPP、＋dp/dtmax、 dp/dtmax、LVSP、LVDP降低，使LVEDP升高，CF增加。选择性大麻素CB1受体拮抗药AM251(1 μmol·L 1)可阻断 Ana的部分心脏效应；另一选择性大麻素CB2受体拮抗药AM630(1 μmol·L 1)对Ana的心脏效应无显著影响。NOS抑制药L N 硝基精氨酸甲酯（L NAME）(100 μmol·L 1)对Ana的心脏效应也无显著影响。Ana能增强原生型NOS(cNOS)活性，抑制诱生型NOS(iNOS)活性，促进心肌NO的释放。结论 Ana使离体大鼠心肌收缩力降低，心率减慢，表现出负性肌力和负性频率作用；Ana可舒张冠脉，增加冠脉流量；大麻素CB2受体可能不参与Ana的这些心脏效应，内源性NO也可能不参与调节Ana的心脏效应；可能存在其他新的作用位点调节Ana的心脏效应。Ana调节心肌NOS同工酶活性，增加cNOS活性，降低iNOS活性，促进NO的释放，可能发挥心肌保护作用，在心肌缺血和高血压治疗中有潜在应用前景。     

Biology,Chemistry and Therapeutic Applications of Nitric Oxide: First International Conference

The First International Conference on the Biology, Chemistry and Therapeutic Applications of Nitric Oxide represented a milestone in the history of nitric oxide (NO) research. This meeting combined the two major conferences on NO, the Biology of Nitric Oxide and the Biochemistry and Molecular Biology of Nitric Oxide. The conference was held at the Hyatt Regency Hotel in San Francisco under the auspices of the Nitric Oxide Society. This meeting successfully brought together scientists from all disciplines currently working in the field. There were three sessions on each day of the meeting, one session for each of the three areas (biology, chemistry and therapy). Each session consisted of two plenary lectures of 20 min duration followed by a series of short 10 min papers, with a total of 70 oral presentations. The meeting also included approximately 400 excellent poster presentations and a hot topics session. All the presentations and posters were of a very high standard and the conference chairs, J Parkinson and G Rubanyi and their co-chairs and scientific advisory board are to be congratulated on such an excellent programme. This review cannot begin to attempt to cover all the contributions, but only to convey the major themes and overall enthusiasm of the meeting. The interested reader is referred to the complete abstracts of the meeting which are published in Nitric Oxide, the journal of the Nitric Oxide Society [1].     

Selective Acetamidine-Based Nitric Oxide Synthase Inhibitors: Synthesis,Docking, and Biological Studies

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Tat-Mediated p66shc Transduction Decreased Phosphorylation of Endothelial Nitric Oxide Synthase in Endothelial Cells

We evaluated the role of Tat-mediated p66shc transduction on the activation of endothelial nitric oxide synthase in cultured mouse endothelial cells. To construct the Tat-p66shc fusion protein, human full length p66shc cDNA was fused with the Tat-protein transduction domain. Transduction of TAT-p66shc showed a concentration- and time-dependent manner in endothelial cells. Tat-mediated p66shc transduction showed increased hydrogen peroxide and superoxide production, compared with Tat-p66shc (S/A), serine 36 residue mutant of p66shc. Tat-mediated p66shc transduction decreased endothelial nitric oxide synthase phosphorylation in endothelial cells. Furthermore, Tat-mediated p66shc transduction augmented TNF-α-induced p38 MAPK phosphorylation in endothelial cells. These results suggest that Tat-mediated p66shc transduction efficiently inhibited endothelial nitric oxide synthase phosphorylation in endothelial cells.     

蝶啶类一氧化氮合酶抑制剂的研究

综述了近年来对一氧化氮化合酶（NOS）结构的研究近况和以四氢生物蝶呤（BH4)为靶点的蝶啶类NOS抑制剂的研究进展，重点讨论了在蝶啶的2－，4－，5－，6－，7－位进行修饰所得的化合物的构效关系。     

三七总皂苷对小鼠柯萨奇B3病毒性心肌炎诱导型一氧化氮合酶的影响

目的：探讨三七总皂苷对小鼠柯萨奇B3病毒所致病毒性心肌炎模型中诱导型一氧化氮合酶（iNOS）表达的影响,为病毒性心肌炎的治疗寻找新的理论依据。方法：选用近交系4～6周龄雄性BALB／C小鼠84只,随机分为3组：空白对照组（C组）24只,心肌炎组（V组）30只,治疗组（T组）30只。于病毒接种后第8天、第15天、第21天分别采血后处死小鼠并留取心脏标本,采用免疫组化法检测心肌iNOS蛋白表达,硝酸还原酶法测定血清NO含量,光镜及透射电镜观察心肌组织病理及细胞超微结构改变。结果：（1）V组和T组小鼠心肌细胞iNOS免疫组化染色表达呈阳性,c组iNOS免疫组化染色阴性;（2）T组与V组比较,心肌细胞iNOS表达在第15天及第21天明显降低,差异有统计学意义（P〈0．05、P〈0．01）;（3）T组在第15天及第21天血清NO含量较V组明显降低（P〈0．05）;（4）光镜下T组炎症细胞浸润较V组轻,炎症积分在第15天及第21天明显低于V组（P〈0．05、P〈0．01）;（5）电镜下V组及T组可见肌原纤维溶解断裂,线粒体膜消失,含溶酶体丰富的单核巨噬细胞浸润,c组小鼠心肌未见上述变化。结论：三七总皂苷可降低小鼠病毒性心肌炎iNOS的表达,从而减少NO释放,可能对病毒性心肌炎有一定治疗作用。     

Drug Discovery for Overcoming Chronic Kidney Disease (CKD): The Endothelin ETB Receptor / Nitric Oxide System Functions as a Protective Factor in CKD

Accelerated cardiovascular disease (CVD) is a frequent complication of renal disease. Chronic kidney disease (CKD) develops hypertension and dyslipidemia, which in turn can contribute to the progression of renal failure. There is general agreement that endothelin-1 (ET-1), which acts through the two subtypes of receptor ET_A and ET_B, plays important physiological roles in the regulation of normal cardiovascular function and that excessive ET-1 production is linked to CVD and CKD. Although selective ET_A or nonselective ET_A/ET_B receptor antagonisms have been recognized as a potential strategy for treatment of several cardiovascular disease, it remains unclear which of the antagonisms is suitable for the individuals with CKD because upregulation of the nitric oxide (NO) system via ET_B receptor is responsible for renal function such as natriuresis, diuresis, and glomerular hemodynamics. Our findings clearly indicate that the blockade of ET receptors, in particular ET_A-receptor antagonism, not only produces a potential renoprotective effect in CKD but also reduces the risk of CVD. In contrast, pharmacological blockade or genetic deficiency of ET_B receptor seems to aggravate CKD and CVD in several experimental models of rats. Moreover, preliminary evidence in patients with CKD also suggests that both selective ET_A- and nonselective ET_A/ET_B-receptor blockade decreases blood pressure but that selective ET_A blockade has additional desirable effects on renal hemodynamics. Thus, at least in CKD, these findings support the notion that ET_B receptor– mediated actions produce a renoprotective effect and that nonselective ET_A/ET_B-receptors blockade seem to offer no advantage over selective ET_A antagonism, and if anything may potentially reduce the benefits.     

1,25-二羟维生素D_3对脊柱结核患者外周血单个核细胞的NO及iNOS的影响研究

目的:研究1,25-二羟维生素D3(DHVD3)作用下脊柱结核患者外周血单个核细胞(PBMC)中一氧化氮(NO)及诱导型一氧化氮合酶(iNOS)含量变化,探讨其对脊柱结核患者的免疫调节作用。方法:患者PBMC分为试验组(加入DHVD3)和对照组(加入培养基),接种卡介苗后培养;采用硝酸还原酶法测定不同时间点(0、3、6、9、12d)试验组和对照组培养上清液中NO的相对吸光度值,分析其变化规律;于第6天收获2组培养细胞,提取细胞总RNA,逆转录后,定量聚合酶链反应(PCR)扩增,对目的基因iNOS及参照基因3-磷酸甘油醛脱氢酶的mRNA进行分析,用相对定量方法2-ΔΔCt比较其差异。结果:细胞培养第6天,试验组NO的相对吸光度值和iNOS的mRNA含量均高于对照组(P<0.05)。结论:DHVD3可使脊柱结核患者PBMC内iNOS的含量增加,进而促进NO的分泌,从而增强单个核细胞对结核杆菌的吞噬作用。     

Vasorelaxing Activity of Ulmus davidiana Ethanol Extracts in Rats: Activation of Endothelial Nitric Oxide Synthase

Ulmus davidiana var. japonica Rehder (Urticales: Ulmaceae) (UD) is a tree widespread in northeast Asia. It is traditionally used for anticancer and anti-inflammatory therapy. The present study investigated the effect of an ethanol extract of UD on vascular tension and its underlying mechanism in rats. The dried root bark of UD was ground and extracted with 80% ethanol. The prepared UD extract was used in further analysis. The effect of UD on the cell viability, vasoreactivity and hemodynamics were investigated using propidium iodide staining in cultured cells, isometric tension recording and blood pressure analysis, respectively. Low dose of UD (10~100μg/ml) did not affect endothelial cell viability, but high dose of UD reduced cell viability. UD induced vasorelaxation in the range of 0.1~10μg/ml with an ED(50) value of 2μg/ml. UD-induced vasorelaxation was completely abolished by removal of the endothelium or by pre-treatment with L-NAME, an inhibitor of nitric oxide synthase. UD inhibited calcium influx induced by phenylephrine and high K(+) and also completely abolished the effect of L-NAME. Intravenous injection of UD extracts (10~100 mg/kg) decreased arterial and ventricular pressure in a dose-dependent manner. Moreover, UD extracts reduced the ventricular contractility (+dP/dt) in anesthetized rats. However, UD-induced hypotensive actions were minimized in L-NAME-treated rats. Taken together, out results showed that UD induced vasorelaxation and has antihypertensive properties, which may be due the activation of nitric oxide synthase in endothelium.