一氧化氮与吗啡镇痛及其镇痛耐受关系的研究进展

近年来,针对吗啡镇痛耐受已经开展了许多研究,通常认为其发生机制非常复杂,涉及许多生物因子和神经传导通路,并且其中的许多因素和环节尚未被完全阐明.一氧化氮(nitric oxide,NO)是中枢神经系统和周围神经系统细胞间和/或细胞内的一种递质,研究发现它在吗啡镇痛作用及其镇痛耐受中发挥着重要调制作用.对此方面的研究进展进行综述,旨在为临床上解决吗啡镇痛耐受这一难题提供有益的启示.     

吗啡耐受形成过程中氯胺酮对小鼠脊髓诱导型一氧化氮合酶mRNA表达的影响

目的观察吗啡耐受形成过程中N-甲基-D天冬氨酸受体拮抗剂氯胺酮对小鼠脊髓诱导型一氧化氮合酶(iNOS)mRNA表达的影响。方法昆明种小鼠30只,随机分为5组(n=6):对照组(A 组)皮下注射生理盐水(10 ml·kg-1)30 min后腹腔注射生理盐水(10 ml·kg-1);慢性吗啡耐受组(B组) 皮下注射吗啡10 mg·kg-1后30min腹腔注射生理盐水10 ml·kg-1;C、D、E组吗啡用药均同B组,注射吗啡后30 min分别腹腔注射5、10、20 mg·kg-1氯胺酮。上述各组每日用药2次,连续9 d。最后1次注射吗啡后1 h处死小鼠,提取腰膨大部位脊髓组织中总RNA,以β-actin为内对照,用半定量反转录聚合酶链反应技术测定iNOS mRNA的表达量。结果 A组未见iNOS mRNA表达,其他各组可见iNOS mRNA表达;与B组相比,C、D、E组iNOS mRNA表达减少(P<0．05);与C组相比,D、E组iNOS mRNA 表达减少(P<0．05)。结论氯胺酮拮抗小鼠吗啡耐受形成与其下词脊髓iNOS mRNA表达有关。     

阿片受体与吗啡镇痛耐受

长期使用阿片类物质如吗啡的主要限制在于生理耐受性的发生.耐受使吗啡的镇痛效果明显减弱.现从阿片受体下调、失敏感、G-蛋白下游信号转导等方面,将阿片受体参与吗啡镇痛耐受机制的研究进展作一综述.     

一氧化氮合酶与急性缺血性肾损伤

一氧化氮合酶是一类复杂的生物合酶，其产物一氧化 机体内发挥广泛而重要的作用。随着对一氧化氮合酶的结构及功能认识的深入，发现一氧化氮及其合酶的变化在急性肾缺血性损伤中有重要作用。     

一氧化氮合酶与急性缺血性肾损伤

一氧化氮合酶是一类复杂的生物合酶，其产物一氧化氮在机体内发挥广泛而重要的作用。随着对一氧化氮合酶的结构及功能认识的深入，发现一氧化氮及其合酶的变化在急性肾缺血性损伤中有重要作用     

内皮细胞型一氧化氮合酶与骨质疏松症

一氧化氮涉及许多生理过程,其对骨细胞的功能也有重要作用.研究发现一氧化氮及其合酶,特别是内皮细胞型一氧化氮合酶在参与和促进骨质疏松症的病理生理过程中占有举足轻重的作用;一氧化氮和内皮细胞型一氧化氮合酶作为骨形成及骨吸收过程中调节因子的作用亦受到广泛关注,为临床应用一氧化氮防治骨质疏松症提供了新思路.目前对一氧化氮和内皮细胞型一氧化氮合酶的骨吸收调控机制及在骨质疏松症发生过程中作用的认识仍以推理性为主,有待进一步研究.     

大鼠脑缺血后一氧化氮合酶的时相变化

探讨一氧化氮有脑缺血中的作用。方法 用分光光度法则脑缺血后ＮＯＳ活性，ＲＴＰＣＲ半定量测脑缺血后ｃＮＯＳ，ｉＮＯＳｍＲＮＡ表达水平，结果 脑缺血后１小时ＮＯＳ活性增加，４小时ＮＯＳ活性开始下降，１２小时 ＮＯＳ活性最低；脑缺血后２４小时，４８小时开始ＮＯＳ活性再镒明显升高。脑缺血后１小时，４小时ｃＮＯＳｍＲＮＡ表达增加，１２小时开始下降，２４小时明显下降，直至４８小时，脑缺血后１小时，４小时ｉＮＯ     

一氧化氮、一氧化氮合酶与伴精索静脉曲张不育患者精液参数的关系

目的:探讨一氧化氮(NO)、一氧化氮合酶(NOS)与伴精索静脉曲张(VC)不育患者精液参数之间的关系。方法:根据体格检查和彩色多普勒超声检查选择伴VC的不育患者(组1,n=53),其中临床型和亚临床型分别为21例和32例;同时选择非VC少弱精子症患者(组2,n=29)和正常生育者(组3,n=28)作对照组。采用硝酸还原法分别测定外周血和精浆中NO含量和NOS活性。用计算机辅助精液分析仪测定VC组患者精子密度、活动精子(a+b级精子)和快速前向运动精子百分率。结果:①组1外周血清NO含量和NOS活性与组2及组3相比差异无显著性(P>0.05),但精浆中NO含量和NOS活性组1明显高于其他两组,差异有显著性(P<0.01和P<0.05)。②组1中,随着曲张的精索静脉内径的增加,外周血清和精浆中NO含量和NOS活性均有所上升,但只有精浆中临床型和亚临床型之间相比差异有显著性(P<0.05)。③组1中,随着精子密度和精子活力的下降,外周血清和精浆中NO含量和NOS活性均有上升趋势,且精子密度≥20×106/ml和≤10×106/ml之间,精子活力≥50%和≤25%之间差异有显著性(P均<0.05)。结论:在VC诊断中精浆中NO含量和NOS活性测定较外周血清中更有意义。早期测定精浆NO含量和NOS活性对VC的诊断和治疗具有重要的临床价值。     

冠心病与内皮型一氧化氮合酶基因多态性的关系

目的 探讨内皮型一氧化氮合酶(eNOS)基因-786T/C,4a4b,894G/T等3个多态性位点与冠心病(CAD)发病相关.方法 对146例中国汉族人群CAD患者和113例正常对照进行遗传学分析,应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)和PCR技术分析2个SNP位点即-786T/C和894G/T,以及1个VNTR位点4a4b,检测各位点基因型和等位基因频率,采用HaploView 4.0及SPSS 13.0软件经x2检验比较两组间各位点基因型及等位基因频率的差异.结果 CAD组中eNOS基因-786T/C位点CC基因型频率为2.0%,4a4b位点4a/4a基因型频率为5.4%,对照组eNOS基因-786T/C位点CC基因型频率为0.0%,4a4b位点4a/4a基因型频率为0.9%,差异有统计学意义(P＜0.05).CAD组和对照组在eNOS基因的894G/T位点等位基因和基因型频率分布差异均无统计学意义(P＞0.05).结论 eNOS基因-786T/C和4a4b多态性与中国汉族人群CAD存在关联,C等位基因和4a等位基因可能是CAD发病的危险因素.eNOS基因894G/T位点与CAD发病无明显相关.

Abstract：

Objective To investigate the relationship between the 3 polymorphisms ( -786T/C,4a4b,894G/T) in endothelial nitric oxide synthase (eNOS) gene and coronary artery disease (CAD).Methods 146 patients with CAD and 113 healthy unrelated individuals in a Chinese Han nation were involved.The genotype and allele frequency of each polymorphism of the eNOS gene in these patients and normal controls were examined by using polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) or PCR methods.Genotypes and allele frequency were analyzed by HaploView 4.0 and SPSS13.0 software.Results The frequency of CC genotype of the -786T/C was 2.0%,and that of 4a/4a genotype of the 4a4b was 5.4% in CAD.The frequency of CC genotype of the - 786T/C was 0.0%,and that of 4a/4a genotype of the 4a4b was 0.9% in controls ( P＜0.05 ).There were significant differences in both allele and genotype frequency of -786T/C and 4a4b between CDA group and control group.Between patients with CAD and controls,there were no significant differences in the frequency of the genotypes and alleles of the 894G/T in eNOS gene.Conclusion The - 786T/C and 4a4b polymorphisms of eNOS gene may be associated with CAD.The individuals with C allele of - 786T/C and 4a allele of 4a4b are susceptible to CAD.There is no significant correlation between 894G/T polymorphism in eNOS gene and CAD.     

一氧化氮供体及一氧化氮合酶抑制剂与动脉疾病

自血管内皮衍生舒张因子 (ｅｎｄｏｔｈｅｌｉｕｍ ｄｅｒｉｖｅｄｒｅｌａｘｉｎｇｆａｃｔｏｒ,ＥＤＲＦ)的化学本质被证实是ＮＯ以来[1] ,学者们对ＮＯ进行了大量的研究 ,证实ＮＯ在许多疾病的病理生理过程中具有重要功能。在血管系统研究方面 ,以往主要集中在ＮＯ的减少与冠心病、高血压相关的研究以及较新的冠脉ＰＴＡ术后再狭窄的基因治疗的研究等方面 ,而对ＮＯ在动脉扩张病及周围动脉硬化闭塞症的作用的研究较少。本文仅就ＮＯ供体和抑制剂在动脉瘤及周围动脉硬化闭塞症中的作用及应用的研究进展作一综述。1　ＮＯ生物学简介1.1　Ｎ…     

iNOS/NO对结直肠肿瘤发生发展的影响

一氧化氮（nitric oxide,NO）具有广泛的生物学活性。近年来发现一氧化氮（NO）、一氧化氮合成酶（nitric oxide synthase-2,NOS-2）与结直肠肿瘤的发生、发展密切相关,它与环氧化酶（cy-cloxygenase-2,COX-2）之间存在复杂的调控机制。对NO清除剂、NOS抑制剂和释放NO的非甾体抗炎药的研究为结直肠肿瘤的防治提供了一个思路。     

Activity of nitric oxide synthase in mature and immature human spermatozoa

Nitric oxide (NO) is known to be involved in multiple signal transduction pathways of male germ cells, including sperm capacitation. In somatic cells, NO production was found to be part of apoptosis signalling. The aim of our study was to further clarify the role of NO in spermatozoa by investigation of NO synthase activity with regard to sperm maturity and sperm apoptosis signalling. Semen specimens from 19 healthy donors were subjected to density gradient centrifugation to separate the predominantly mature and immature sperm fraction. NO synthase activity was evaluated using diaminofluoresceine‐2‐diacetate by FACS. Apoptosis signalling was monitored by flowcytometric analyses of caspase‐3 (CP3) and integrity of the transmembrane mitochondrial potential (TMP). TUNEL assay was used to detect DNA fragmentations. Maturity of human spermatozoa was associated with increased NO synthase activity and inactivated apoptosis signalling (lower levels of disrupted TMP, active CP3 and DNA fragmentations, P < 0.05). Activation of apoptosis signalling was significantly negatively correlated to NO production, indicating a rather anti‐apoptotic effect of NO. This might underline the recently proposed role of NO in physiological sperm signal transduction, e.g. during capacitation.     

大鼠肝缺血／再灌注后肝组织一氧化氮和不同亚型一氧化氮合酶的变化

目的探讨一氧化氮（NO）和一氧化氮合成酶（NOS）在肝缺血／再灌注（I／R）过程中的变化和作用。方法健康雄性SD大鼠24只，随机分为3组（每组8只）：①正常对照组，术中只分离肝周围韧带，不做肝门阻断及再灌注。②I/R组，进行45min的部分肝门阻断及60min的再灌注。③L-精氨酸（L—Arg）组，缺血前20min经阴茎背静脉注射L—Arg（300mg／kg），余同②组。实验结束后，取下腔静脉血2ml，并迅速切取缺血肝组织。检测血清丙氨酸转氨酶（ALT）、门冬氨酸转氨酶（AST）、乳酸脱氢酶（LDH）；测定肝组织中超氧化物歧化酶（SOD）、丙二醛（MDA）、黄嘌呤氧化酶（XOD）、一氧化氮（NO）和一氧化氯合成酶（NOS）等指标；观察光镜和电镜下肝组织学变化。结果与正常对照组相比，I／R组iNOS升高，NO降低；L-Arg组NO、eNOS均高于I／R组。2、3组比1组大鼠的肝组织病理损害重、肝功能差，L—Arg组病理损害较I／R组明显减轻、肝功能改善。结论NO对大鼠肝I／R损伤具有保护作用．不同亚型NOS的变化参与其中。     

Expression of nitric oxide and inducible nitric oxide synthase in acute renal allograft rejection in the rat

BACKGROUND: Recent studies have shown that nitric oxide (NO) synthases, particularly inducible nitric oxide synthase (i-NOS), are induced in acute rejection episodes following heart, liver, pancreas and kidney allotransplantation. Furthermore, tissue and cellular injury has been demonstrated to be mediated by peroxynitrite (ONOO-), a metabolite of NO as well as a potent oxidant. However, a detailed relationship between NO, i-NOS and graft injury in transplantation remains elusive. METHODS: The present study used the following models of renal transplantation in rats: allografts (n = 5, Brown-Norway to Lewis [LEW] rats), isografts (n = 5, LEW to LEW) and allografts treated with aminoguanidine (AG), an i-NOS inhibitor (n = 5). Blood urea nitrogen (BUN), serum creatinine (SCr) and urinary and serum nitrosocompounds (NOx) were measured on days 2, 4 and 7 post-transplant. Western blot analysis of i-NOS protein expression and measurement of i-NOS activity were carried out in grafts harvested on Day 7, along with immunohistochemical and histopathological examinations. RESULTS: In the allograft group, both BUN and SCr levels increased markedly on Day 7, in parallel with a sharp increase in NOx. A band stained by anti-i-NOS antibody was detected at approximately 130 kDa, along with high levels of i-NOS activity and diffusely distributed i-NOS-positive cells (macrophages). Histologically, an acute rejection episode was confirmed (Grade 3 according to Banff classifications). In the AG group, reduced renal function and graft injury were significantly less severe than in the allograft group. CONCLUSIONS: In rat renal allograft acute rejection, markedly increased levels of serum NOx were observed, along with enhanced tissue i-NOS activity, together resulting in graft injury. AG administration suppressed the increase of serum NOx levels, with concomitant mitigation of tissue injury and renal function impairment.     

诱导型一氧化氮合成酶在迟发性血管痉挛中的作用

目的　以大鼠迟发性脑血管痉挛模型为基础研究诱导型一氧化氮合成酶 (iNOS)在迟发性血管痉挛发展中的作用。方法　 3 2只雄性SD大鼠随机分为实验组和对照组 ,实验组枕大池二次注血诱导迟发性脑血管痉挛 ,对照组枕大池注射生理盐水。第 8天行脑血管造影 ,枕大池抽取脑脊液测一氧化氮 (NO)浓度。逆转录 聚合酶链反应 (RT PCR )法和免疫组织化学法测定并评价iNOSmRNA和蛋白质在基底动脉、大脑中动脉和皮质中的表达。结果　颅内动脉血管减影提示对照组颈内动脉颅内段、大脑中动脉 (MCA)明显变细 ,大脑中动脉中段直径 (MD)与镫骨动脉中段直径 (SD)之比衡量大脑中动脉的管径显示实验组MCA管径较对照组MCA管径减少 3 0 %。对照组脑脊液中NO浓度为 (11.70± 2 .62 ) μmol/L ,实验组脑脊液中NO的浓度为(5 5 .67± 12 .84)μmol/L。iNOSmRNA和蛋白质表达于基底动脉、大脑中动脉和皮质 ,其中基底动脉表达最强。 结论　iNOS作为迟发性脑血管痉挛发展中的关键因素参与血管壁的迟发性损伤。     

The distribution of neuronal and inducible nitric oxide synthase in urethral stricture formation

PURPOSE: The distribution of neuronal (n) and inducible (i) nitric oxide synthase (NOS) may have a role in the maintenance of normal urethral spongiosum and during the development of spongiofibrosis in urethral stricture disease. MATERIALS AND METHODS: Eight normal and 33 strictured human bulbar urethras were studied by histological and immunohistochemical techniques for the neuronal markers S-100, nNOS and iNOS. The smooth muscle-to-collagen ratio was calculated by morphometric analysis of Masson's trichrome sections. Immunohistochemical staining patterns of the neuronal markers in normal urethral tissue was compared to that in urethral stricture tissue with spongiofibrosis. RESULTS: The smooth muscle-to-collagen ratio was significantly lower in the strictured urethra compared to that in the control group (p = 0.001). In the strictured bulbar urethra nNOS immunoreactivity was decreased compared to that in normal urethral tissue. The severity of spongiofibrosis corresponded to the loss of nNOS immunoreactivity. iNOS immunoreactivity was found in strictured urethral epithelium and spongiosal tissue, whereas the control group was nonimmunoreactive to iNOS. CONCLUSIONS: Urethral stricture formation is a fibrotic process associated with significant changes in NOS metabolism. Abnormal collagen synthesis following urethral trauma may be stimulated by inappropriate iNOS activity. A functional nerve supply to the urethral spongiosum seems to be crucial in the maintenance of the unique ultrastructure of the urethral spongiosum.     

脑出血患者一氧化氮、一氧化氮合酶的变化及临床意义

目的 探讨一氧化氮(NO)、一氧化氮合酶(NOS)与脑出血的关系。方法 测定76例脑出血患者及66例健康人血浆NO、NOS含量，分析上述指标在出血后3个时期的变化以及与患者颅内血肿量、是否合并蛛网膜下腔出血和临床预后的关系。结果 与对照组相比，脑出血1～3d组NO、NOS含量明显高于4～7d组、14d组和对照组，NO、NOS变化与颅内血肿量和／或合并蛛网膜下腔出血有一定关系。结论 NO、NOS参与了脑出血病理生理过程，其变化对病情的预后有一定的意义。     

Iron deficiency suppresses ileal nitric oxide synthase activity

Intestinal motility disorders are more common in women of childbearing age who are prone to iron deficiency anemia. The neurotransmitters nitric oxide (NO) and acetylcholine (ACh) play a key role in ileal smooth muscle relaxation and contraction, respectively. Iron-containing heme is known to be a cofactor for nitric oxide synthase (NOS), the enzyme responsible for NO production. Therefore we tested the hypothesis that iron deficiency would downregulate ileal NOS activity without affecting the ileum’s response to ACh. Twelve adult female prairie dogs were fed either an ironsupplemented (Fe+) (200 ppm) (n = 6) or an iron-deficient (Fe-) (8 ppm) (n = 6) diet for 8 weeks. Ileal circular muscle strips were harvested to measure responses to ACh and electrical field stimulation. Under nonadrenergic noncholinergic (NANC) conditions, Nω-nitro-L-arginine (L-NNA), an NOS inhibitor, and VIP_10-28, a vasoactive intestinal peptide (VIP) inhibitor, were added prior to electrical field stimulation. NANC inhibitory responses are expressed as a percentage of optimal relaxation from EDTA. The excitatory response to ACh was similar in both groups (1.1 ± 0.3 N/cm² vs. 1.5 ± 0.3 N/cm², P = 0.45). The inhibitory response to electrical field stimulation under NANC conditions was greater in the Fe+ group (34.7 ±2.9%) compared to the Fe-group (23.9 ±3.2%; P <0.01). L-NNA eliminated the inhibitory response in the Fe+ group (0.02 ± 0.02%) but not in the Fe-group (8.38 2 2.15%; P <O.Ol). VIP_10-28 led to greater relaxation in the Fe+ animals (45.8 ± 6.6%) than in the Fe-animals (23.4 ±5.8%; P <0.05). Both L-NNA and VIP_10-28 had no inhibitory response (0.02 ± 0.02%) in the Fe+ animals, whereas the Fe-animals had some residual inhibition (2.54 ±1.04%; P <0.05). These data suggest that ileal NANC relaxation is due to NOS and that iron deficiency results in (1) decreased NANC relaxation, (2) a compensatory relaxation due to a non-NOS, non-VIP mechanism, and (3) a normal excitatory response. We conclude that iron deficiency suppresses ileal NOS activity. Supported by grant ROI-DK44279-07 from the National Institutes of Health. Presented at the Forty-First Annual Meeting of The Society for Surgery of the Alimentary Tract, San Diego, Calif., May 2l–24, 2000.