2型糖尿病合并冠心病患者血清脂联素、血皮素-1和一氧化碳与血管内皮功能相关性研究

目的 研究糖尿病合并冠心病患者血清脂联素(APN)、一氧化氮(NO)、内皮素(ET-1)、高密度脂蛋白(HDL)、反应性充血肱动脉内径增加程度(FMD)的水平,并分析其相关性,探讨脂联素对内皮功能的影响.方法 75例糖尿病患者分为单纯糖尿病组(B组,35例)和合并冠心病组(C组,40例),正常对照(A组)33例,分别检测血清APN、NO、ET-1,同时利用超声检测FMD、硝酸甘油诱发内皮独立的肱动脉直径增加程度(NID).结果 B组、C组与A组比较,血APN、NO、HDL显著降低(P＜0.01);FBG、ET-1显著升高(P＜0.01).FMD(%)降低(P＜0.05),NID(%)各组问差异无显著性(P＞0.05).APN与ET-1、FBG、BMI呈负相关(r=-0.41,r=-0.49,r=-0.31,P＜0.01),与FMD(%)、NO、HDL呈显著正相关(r=0.672,r=0.308.r=0.347,P＜0.01).结论 2型糖尿病合并冠心病患者存在明显的内皮功能紊乱,脂联素可通过影响ET-1和NO的释放,改善血管内皮功能.     

NO调控JNKs通路对鼠海马神经元保护作用研究

Background C-Jun N-terminal kinase (JNK) signaling pathway plays a critical role in cerebral ischemia. Although the mechanistic basis for this activation of JNK1/2 is uncertain, oxidative stress may play a role. The purpose of this study was to investigate whether the activation of JNK1/2 is associated with the production of endogenous nitric oxide. Methods Ischemia and reperfusion was induced by cerebral four-vessel occlusion (4-VO). Sprague–Dawley (SD) rats were divided into 6 groups: sham group, ischemia and reperfusion group, neuronal nitric oxide synthase (nNOS) inhibitor 7-Nitroindazole (7-NI) (Sigma St Louis, MO, USA) given group, inducible nitric oxide synthase (iNOS) inhibitor (AMT) (Sigma) given group, sodium chloride control group and 1% dimethyl sulfoxide (DMSO) control group. The levels of protein expression and phospho-JNK1/2 were detected by western blot and the survival hippocampus neurons in CA1 zone were observed by cresyl violet (CV) staining. Results The study illustrated two peaks of JNK1/2 activation occurred at 30 min and 3 days during reperfusion. 7-NI inhibited JNK1/2 activation during the early reperfusion, whereas AMT preferably attenuated JNK1/2 activation during the later reperfusion. At same time, the results also showed that administration of 7-NI and AMT can decrease ischemia/Reperfusion (I/R)-induced neuronal loss in hippocampal CA1 region. Conclusions JNK1/2 activation is associated with endogenous nitric oxide (NO) in response to ischemic insult.     

Effects of Nephritis No.3 Recipe on Nitric Oxide,Nitric Oxide Synthase Secreted by Cultured Mesangial Cells in Rats and the Gene Expression of Inducible Nitric Oxide Synthase

Objective:To explore the effect of the Nephritis No.3(N-3)recipe on nitric oxide(NO),nitric oxide synthase(NOS)secreted by cultured mesangial cells(MC)and its gene expression of the in-ducible nitric oxide synthase(iNOS).Methods:The drug(nephritis No.3)-containing serum was preparedwith serum pharmacological technique,and then was applied to react on mesangial cells cultured In fetalcalf serum(FCS)and cells cultured in FCS plus lipopolysaccharide.To observe the secretion of NO andNOS and the gene expression of iNOS by means of RT-PCR.Results:Under the two kinds of culture con-ditions,the content of NO and NOS in the groups with drug-containing serum were higher than thosewithout drug-containing serum(P<0.05,P<0.01),and the expression of iNOS mRNA was up-regula-ted too.Conclusion:The N-3 could significantly promote the secretion of NO and NOS and the mRNA ex-pression of iNOS in rats.     

Activation of c-Jun N-terminal kinase 1/2 regulated by nitric oxide is associated with neuronal survival in hippocampal neurons in a rat model of ischemia

Background  C-Jun N-terminal kinase (JNK) signaling pathway plays a critical role in cerebral ischemia. Although the mechanistic basis for this activation of JNK1/2 is uncertain, oxidative stress may play a role. The purpose of this study was to investigate whether the activation of JNK1/2 is associated with the production of endogenous nitric oxide (NO).

Methods  Ischemia and reperfusion (I/R) was induced by cerebral four-vessel occlusion. Sprague-Dawley (SD) rats were divided into 6 groups: sham group, I/R group, neuronal nitric oxide synthase (nNOS) inhibitor (7-nitroindazole, 7-NI) given group, inducible nitric oxide synthase (iNOS) inhibitor (2-amino-5,6-dihydro-methylthiazine, AMT) given group, sodium chloride control group, and 1% dimethyl sulfoxide (DMSO) control group. The levels of protein expression and phospho-JNK1/2 were detected by Western blotting and the survival hippocampus neurons in CA1 zone were observed by cresyl violet staining.

Results  The study illustrated two peaks of JNK1/2 activation occurred at 30 minutes and 3 days during reperfusion. 7-NI inhibited JNK1/2 activation during the early reperfusion, whereas AMT preferably attenuated JNK1/2 activation during the later reperfusion. Administration of 7-NI and AMT can decrease I/R-induced neuronal loss in hippocampal CA1 region.

Conclusion  JNK1/2 activation is associated with endogenous NO in response to ischemic insult.

     

雌激素对血管内皮细胞一氧化氮合酶活性的调控

目的　观察 17β -雌二醇对大鼠血管内皮细胞一氧化氮合酶 (NOS)活性的影响。 方法　用贴壁法和无酚红 164 0培养基培养大鼠血管内皮细胞 ,在 10nmol的 17β -雌二醇作用下 ,观察一定时间内内皮细胞的NOS活性。 结果　 10nmol的 17β -雌二醇作用 8～ 2 4hNOS活性显著增强 (同对照组相比 8hP <0 0 5 ;16h ,2 4hP <0 0 1)。 结论　 17β-雌二醇能增强大鼠血管内皮细胞的NOS活性 ,这可能是雌激素降低血管阻力、抑制动脉粥样硬化作用的重要机理之一     

Modulation of ciliary activity by tumor necrosis factor-alpha in cultured sinus epithelial cells. Possible roles of nitric oxide

The primary function of well-differentiated ciliated epithelium in the paranasal sinus is to eliminate harmful agents through the beating action of cilia. Respiratory epithelium also contributes to local inflammatory processes through the release of various proinflammatory cytokines. Recently, considerable attention has been focused on the intimate relationship between the cytokine-dependent regulation of the ciliary beat frequency (CBF) and intra-cellular production of nitric oxide (NO) in ciliated epithelial cells. The aims of this study are to examine the effect of tumor necrosis factor-alpha (TNF-alpha), one of the major proinflammatory cytokines, on the ciliary activity of human sinus epithelial cells and to assess the hypothesis that NO is involved in this regulatory mechanism. Human maxillary or ethmoidal sinus mucosa (n = 23) were cultured by the explant-outgrowth method. CBF of the outgrowth ciliated cells was measured by the photoelectrical method before and after being treated with TNF-alpha (0.1, 1 and 10 ng/ml) or dexamethasone (10(-6) M and 10(-7) M). We also investigated the expression of nitric oxide synthase (NOS) isoforms, enzymes responsible for NO synthesis, by fluorescent immunohistochemistry. TNF-alpha increased CBF at relatively low concentrations (0.1 and 1 ng/ml) and decreased CBF at a high concentration (10 ng/ml). Dexamethasone decreased CBF at a concentration of 10(-6) M. Fluorescent immunohistochemistry demonstrated that the expression of inducible NOS was augmented by TNF-alpha and attenuated by dexamethasone, whereas that of endothelial NOS remained unchanged. We conclude that human sinus epithelial cells potentially contribute to the inflammatory process by regulating their ciliary motility through an NO-dependent pathway. Proinflammatory cytokines and steroids are able to modulate this mechanism by the induction or inhibition of expression of different NOS isoforms.     

铅对大鼠海马一氧化氮合酶活性的影响

目的：探讨铅对海马LTP的影响与海马不同亚区NOS活性变化的关系。方法：采用NADPH—d组化法对铅暴露大鼠海马不同亚区内一氧化氮合酶活性的变化进行研究。结果：染铅大鼠海马CAI区和齿状回NOS阳性细胞数均明显少于对照组（P＜0．05）;在CA3区染铅组NOS阳性细胞数目与对照组无明显差别。结论：推测铅对海马LTP的影响可能与染铅后海马各区NOS活性的不同变化有关。     

内皮型一氧化氮合酶活性的调节与心血管疾病的研究进展

内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)是合成一氧化氮(nitiric oxide,NO)起主要作用的酶,eNOS和NO在调节血管壁结构和功能中有重要作用,参与心血管疾病的病理生理过程。eNOS活性的改变除了常见的磷酸化途径外,还涉及乙酰化、谷胱甘肽化、蛋白质间的相互作用等机制,笔者将简要阐述eNOS的基本结构和功能、eNOS活性的调节途径及其在心血管疾病中的研究进展。     

Propofol improves cardiac functional recovery after ischemia-reperfusion by upregulating nitric oxide synthase activity in the isolated rat hearts

Background There are few studies to assess whether propofol attenuates myocardial ischemia-reperfusion injury via a mechanism related to nitric oxide （NO） route, so we designed this randomized blinded experiment to observe the changes of NO contents, nitric oxide synthase （NOS） activity, NOS contents in the myocardium, and cardiac function in ischemic reperfused isolated rat hearts, and to assess the relation between myocardial NO system and cardioprotection of propofol. Methods The hearts of 30 Sprague-Dawley male rats were removed, mounted on a Langendorff apparatus, and randomly assigned to one of three groups （n=10 each group） to be treated with the following treatments in a blinded manner： Group 1, control group, after perfusion with pure Krebs Henseleit bicarbonate （K-HBB） buffer solution for 15 minutes, hearts were subjected to 20 minutes global ischemia followed by 60 minutes reperfusion with pure K-HBB buffer; Group 2, after perfusion with K-HBB buffer solution containing propofol （10 ug/ml） for 15 minutes, the hearts underwent 20 minutes global ischemia followed by 60 minutes reperfusion with the same K-HBB buffer solution; Group 3, after perfusion with K-HBB buffer solution containing propofol （10ug/ml） and L-NAME （100 umol/L） for 15 minutes, the hearts underwent 20 minutes global ischemia followed by 60 minutes reperfusion with the same K-HBB buffer solution. The cardiac function was continuously monitored throughout the experiment. The coronary flow was also measured. An ISO-NO electrode was placed into the right atrium close to the coronary sinus to continuously measure NO concentration in the coronary effluent. The tissue samples from apex of hearts in Groups 1 and 2 were obtained to measure the NOS activity by spectrophotometry and the NOS contents by immunohistochemistry, respectively. Results The cardiac function was significantly inhibited after ischemia and then gradually improved with reperfusion in all three groups. As compared with Group 1, the cardiac function variables and coronary flow at all the observed points were significantly improved in Group 2. The cardiac function variables and coronary flow were better in Group 3 than in Group 1, but were inferior in Group 3 than in Group 2. Both NO contents and NOS activity in the myocardium were significantly higher in Group 2 than in Group 1. However, NOS contents in the myocardium did not significantly differ between Groups 1 and 2. Conclusions In isolated rat hearts, propofol can improve cardiac functional recovery after ischemia-reperfusion by upregulating NOS activity in the myocardium. The NO system may play an important role in the preservation of myocardial ischemia-reperfusion injury produced by propofol.     

甲基苯丙胺对大鼠纹状体小胶质细胞及NOS的影响

目的 观察甲基苯丙胺(METH)对大鼠纹状体内小胶质细胞的影响以及一氧化氮合酶(NOS)、诱导型一氧化氮合酶(iNOS)和结构性一氧化氮合酶(cNOS)的变化以及相互间的关系.方法 建立METH给药模型,实验组给予METH,对照组给予等体积生理盐水,采用OX-42免疫组化观察小胶质细胞的变化情况,采用酶化学方法测量纹状体内的NOS、iNOS和cNOS的活性改变.结果 与对照组相比,实验组纹状体区小胶质被细胞激活,数量显著增加(P<0.01),呈现为"灌木丛"(bushy)甚至"阿米巴样"(amoeboid),同时NOS、iNOS和cNOS的活性均显著升高(P<0.01).结论 小胶质细胞被激活,NOS活性升高,可能是METH引起中枢神经损伤的重要机制.     

肺癌患者肺泡巨噬细胞一氧化氮活性研究

用镀铜镉还原－Ｃｒｉｅｓｓ法检测肺癌患者ＢＡＬＦ及肺泡巨噬细胞（ＡＭ）培养上清液中ＮＯ水平；ＲＴ－ＰＣＲ检测ＡＭ ｉＮＯＳ ｍＲＮＡ表达。结果显示，肺癌组ＢＡＬＦ及ＡＭ培养上清液中ＮＯ水平均明显低于对照组。两组ＡＭ ｉＮＯＳ ｍＲＮＡ表达阳性率分别为６９％和９１％（Ｐ〉０．０５），但表达强度肺癌组明显弱于对照组（Ｐ〈０．０１）。经ＧＭ－ＣＳＦ刺激后，ＡＭ ｉＮＯＳ表达强度及培养上清液中ＮＯ水平均显     

Inducible nitric oxide synthase is involved in the oxidation stress induced by HIV-1 gp120 in human retina pigment epithelial cells

Background The human immunodeficiency virus-1 （HIV-1） envelope glycoprotein gp120 has been implicated in the development of AIDS-associated retinopathy. The present study tested the hypothesis that gp120 may induce oxidative stress including up regulation of inducible nitric oxide synthase （iNOS） and production of malondialdehyde （MDA） and nitric oxide （NO） to mediate retinopathy in retinal pigment epithelial （RPE） cells. Methods Human RPE cell line D407 was cultured and treated with gp120. HIV-1 gp120 protein induced lipid peroxidation product MDA. NO production and iNOS expression were examined in vitro by spectrophomtometry, real-time PCR, Western blotting, and confocal microscope. Results Addition of gp120 was able to induce RPE cells to produce NO and MDA in time- and dose-dependent manners （P 〈0.05）. Similarly, gp120 was also capable of up-regulating iNOS mRNA and protein in D407 cells in time- and dose-dependent manners. Conclusions Gp120 induces oxidative stress in D407 cell by stimulating MDA and NO production, which is mediated by up-regulating iNOS expression. Gp120 may mediate oxidation stress in AIDS-associated retinopathy.     

呼出气一氧化氮测定对儿童支气管哮喘的诊断价值

目的:评估呼出气一氧化氮(feno)测定对儿童支气管哮喘诊断的临床价值?方法:选取2010年6月~9月就诊于南京市儿童医院疑似支气管哮喘的患儿87例,测定feno水平,以临床表现和支气管激发或舒张试验为哮喘诊断的金标准,绘制roc曲线,结合roc曲线明确feno的诊断临界点,以评价feno对儿童支气管哮喘的诊断价值?结果:87例患儿中诊断哮喘患儿52例,非哮喘患儿35例;哮喘组feno[(59.77±42.48)ppb]高于非哮喘组[(25.26±22.60)ppb,p < 0.05]?fev1%与feno无直线相关(r=-0.151,p > 0.05)?feno与外周血嗜酸性粒细胞有相关性(r=0.546,p < 0.01)?roc曲线下面积是0.818,feno以34.5 ppb为阈值,敏感度0.712,特异度0.686,阳性预测值0.755,阴性预测值0.605,正确率0.689?结论:feno测定有助于儿童支气管哮喘的诊断和鉴别诊断?     

帕金森病大鼠中枢神经一氧化氮合成酶活性研究

目的 通过对帕金森病(Parkinson's disease,PD)大鼠中枢神经系统一氧化氮合成酶(NOS)3种同工酶的活性变化的研究,探讨帕金森病发病的可能因素和机制.方法 用6-羟多巴胺(6-OHDA)毁损单侧黑质建立大鼠向健侧旋转运动的帕金森病动物模型;采用分光光度法检测神经元型一氧化氮合成酶(nNOS)、内皮型...     

异丙酚对脂多糖诱导人脐静脉内皮细胞内皮源型一氧化氮合酶基因启动子转录活性的影响

目的 探讨异丙酚对脂多糖(LPS)诱导人脐静脉内皮细胞(HUVEC)内皮源型一氧化氮合酶(heNOS)基因启动子转录活性的影响.方法 以基因重组技术构建heNOS基因启动子区(-1～-1600 bp)驱动的萤火虫荧光素酶报告基因载体pGL2-Basic,得到质粒peNOS-Luc.采用脂质体介导的细胞基因共转染技术将peNOS-Luc、空载体pGL2-Basic和?-半乳糖苷酶表达质粒pCMV-?共转染HUVEC,用LPS、LPS 异丙酚和LPS 转移生长因子?1(TGFb1)分别刺激转染后的HUVEC,检测并比较荧光素酶/?-半乳糖苷酶活性,以确定LPS、异丙酚和TGF?1对heNOS基因启动子转录活性的影响.结果 (1)酶切和测序结果均证实重组载体peNOS-Luc构建正确;(2)重组载体peNOS-Luc在HUVEC中有效表达;(3)与未加刺激组比较,LPS刺激组heNOS启动子的转录活性降低.与LPS刺激组比较,LPS 异丙酚和LPS TGF?1组heNOS启动子的转录活性增强.结论 异丙酚能明显增强heNOS基因启动子的转录活性,可初步证实异丙酚在转录水平通过上调heNOS基因启动子的转录活性而影响NO的生成和释放,这可能是异丙酚防治LPS诱导引起炎症反应的机制之一.     

高血压病患者血浆氧自由基、纤溶活性、一氧化氮的变化及丹参粉针干预作用

目的 探讨高血压病患者氧自由基、纤溶活性、一氧化氮的变化及丹参粉针干预作用.方法 ①观察80例正常人和80例高血压病患者血浆丙二醛(MDA)、超氧化物歧化酶(SOD)、组织型血纤维蛋白溶酶原激活剂(t-PA),t-PA抑制物(PAⅠ)及一氧化氮(NO),并比较两组MDA、SOD、t-PA、PA Ⅰ及NO的变化;②80例高血压组随机分为对照组40例和观察组40例,对照组予左旋氨氯地平,观察组予左旋氨氯地平联合丹参粉针治疗,比较两组治疗前后MDA、SOD、t-PA、PA Ⅰ及NO的变化.结果 ①高血压组的MDA及PA Ⅰ较正常人组明显升高,而SOD、t-PA和NO较正常人组明显降低(P＜0.05).②对照组治疗前后MDA、SOD、t-PA、PA Ⅰ和NO的变化差异无统计学意义(P＞0.05),观察组治疗后MDA和PA Ⅰ较治疗前明显下降(P＜0.01),而SOD、t-PA、NO明显升高(P＜0.01).结论 高血压病患者存在MDA、SOD、t-PA、PA Ⅰ及NO异常,丹参粉针能提高患者的SOD活力,降低MDA含量,促进纤溶活性,NO合成释放增多.     

脑缺血后大鼠蝶腭神经节一氧化氮合酶表达的实验研究

目的　探讨一氧化氮合酶 (NOS)在大鼠蝶腭神经节内神经元中的表达与缺血再灌注脑损伤的关系。方法　 48只SD雄性大鼠 ,采用Pulisislli法制作闭塞大鼠四动脉的全脑缺血模型后随机分为实验组和假手术对照组 ,采用还原型尼克酰胺嘌呤二核苷酸脱氢酶 (NADPH d)组化法显示NOS阳性神经元 ,观察两组蝶腭神经节内NOS阳性神经元细胞数目的变化。结果　再灌注后NOS在蝶腭神经节内神经元表达水平增高 ,与假手术对照组相比 ,差异有极显著意义 (P <0 .0 1)。结论　NOS在大鼠缺血再灌注后蝶腭神经节内神经元中表达水平的增高 ,可能是缺血性脑损伤后的一种自身保护性调节反应