血流切应力调控内皮型一氧化氮合酶的分子机制

血流切应力（flowshear stress，rss）是生理或病理条件下调节血管内皮细胞产生一氧化氮的最重要的刺激因素。FSS对内皮型一氧化氮合酶（endothelial nitric oxide synthase，eNOS）的调控包括基因转录的调节、转录后的调节和翻译后的调节。eNOS基因转录以及转录后mRNA的稳定性能被FSS诱导加强。FSS通过游离钙离子浓度、磷酸化、eNOS相关蛋白以及细胞内易位等途径调节eNOS的催化活性。此外，FSS还能调控eNOS催化反应的辅助因子。     

Activation of human neutrophils by the pollutant sodium sulfite: effect on cytokine production,chemotaxis, and cell surface expression of cell adhesion molecules

In chronic beryllium disease (CBD), a granulomatous lung disease characterized by hypersensitivity to beryllium salts (BE), BE challenge of bronchoalveolar lavage cells induces IFNgamma. Although nitric oxide (NO) is elevated in CBD airways, the effects of NO on CBD IFNgamma responses are unknown. Here we report that BE-stimulated IFNgamma production in CBD lavage cells was markedly reduced (74%) by the NO generator DETA NONOate. Investigation of IFNgamma-stimulatory cytokine involvement indicated that lavage cell IL-18 was significantly increased (fourfold) by BE and reduced (64%) by DETA NONOate but IL-12 was undetectable. IL-18 production was caspase-1-dependent but caspase 1 inhibition reduced IFNgamma only partially (43%). Specific antibody depletion of lavage cell IL-18 yielded marginal reduction (19%) of IFNgamma. Data are the first to show that: (1) BE stimulates IL-18 as well as IFNgamma in CBD; (2) BE cytokine responses are NO-sensitive; and (3) NO down-regulation of IFNgamma involves other sites in addition to IL-18.     

大鼠脑干神经元型一氧化氮合酶免疫阳性神经元的分布

目的　观察大鼠脑干神经元型一氧化氮合酶 (nNOS)免疫阳性神经元的分布 ,为探讨nNOS的作用提供形态学资料。方法　用ABC免疫细胞化学方法显示脑干nNOS免疫阳性神经元。结果　大鼠脑干nNOS免疫阳性神经元以中脑和脑桥分布丰富 ,延髓较稀少 ;在中脑 ,nNOS免疫阳性神经元主要分布于中脑水管周围灰质的背侧部、被盖背外侧核、中缝背核、上下丘灰质等部位 ;在脑桥 ,主要分布于被盖背外侧核、脑桥中缝核、被盖脚桥核、蓝斑、臂旁核、斜方体核 ,以及脑桥网状结构 ;与中脑和脑桥相比 ,延髓nNOS免疫阳性神经元较少 ,主要分布于延髓网状结构、三叉神经脊束核和孤束核等核团。结论　分布于脑干内丰富的nNOS免疫阳性神经元可能通过其生成的NO调节其他神经递质的分泌 ,共同参与内脏活动、感觉和运动的传导 ,以及睡眠和觉醒等脑的高级整合功能的调节。     

Inhalation anesthetic-induced neuroinvasion by an attenuated strain of West Nile virus in mice

There are contradictory reports regarding the effects of inhalation anesthetics on the immune system. Measurable immune responses have been studied in vitro, but little is known about the in vivo effects in the intact organism. We used an attenuated, non-neuroinvasive, nonlethal strain of the encephalitic West Nile virus, termed WN-25, which can become lethal in combination with environmental stressors, to study possible modulatory immune effects of inhalation anesthetics in mice. Both single short-term exposure and repeated exposure to halothane and nitrous oxide were studied. Exposure to 30% CO2 served as a positive control. Mortality, brain invasion, spleen weight, and antiviral antibodies served as the experimental endpoints. Halothane and nitrous oxide led to viral brain invasion, increased mortality, and suppressed immune response in a concentration- and time-dependent manner. Repeated exposures had a cumulative effect. Assessment of the stability of the viral attenuation did not demonstrate any alteration in the character of the virus, suggesting an increased access to the brain by inhalation anesthetics that led to the fatal encephalitis. These findings may be of special concern to populations at risk, such as operating room staff and patients undergoing general anesthesia in endemic areas of encephalitic virus species, in which subclinical infection may develop into an overt disease.     

缺氧性肺动脉高压时一氧化氮合酶、左旋精氨酸及亚硝基左旋精氨酸甲酯的研究

目的研究一氧化氮（ＮＯ）在缺氧性肺动脉高压（ＨＰＨ）发病中的作用。方法用烟酰胺腺嘌呤二核苷酸磷酸—黄递酶（ＮｉｃｏｔｉｎａｍｉｄｅＡｄｅｎｉｎｅＤｉｎｕｃｌｅｏｔｉｄｅＰｈｏｓｐｈａｔｅＤｉａｐｈｏｒａｓｅ，ＮＡＤＰＨｄ）和免疫组化ＡＢＣ方法检测原生型和诱生型一氧化氮合酶（ｃＮＯＳ、ｉＮＯＳ）在正常和缺氧大鼠肺内的表达和分布，同时观察左旋精氨酸（Ｌａｒｇ）和亚硝基左旋精氨甲酯（ＬＮＡＭＥ）对正常和缺氧大鼠肺循环的影响。结果ＨＰＨ组，ＮＡＤＰＨｄ阳性反应见于大血管内皮、平滑肌、支气管粘膜上皮及小血管内皮和平滑肌中，而后者在正常时未见阳性反应；ｃＮＯＳ在肺血管内皮和支气管粘膜上皮中的表达明显减弱，甚至消失，而正常时不表达ｉＮＯＳ的肺血管内皮和血管、支气管平滑肌在ＨＰＨ组出现了阳性表达；缺氧时补充Ｌａｒｇ和ＬＮＡＭＥ，与单纯缺氧相比，对右心室肥大和肺血管重建无影响。结论缺氧时ｃＮＯＳ被抑制，可能对ＨＰＨ的形成具有一定的作用；而ｉＮＯＳ的诱导表达，则可能对ＨＰＨ的形成具有阻止作用。     

Expression of inducible nitric oxide synthase induced by lipid-associated membrane proteins of Ureaplasma urealyticum is regulated by nuclear factor κB-mediated mechanism in murine macrophages

U.urealyticumisthesmallestprokaryoticorgan ismcapableofself replication.Thetinymicroor ganismcouldbeisolatedfromurogenital,placen tasandtherespiratorytractsofpreterminfants.Moreover,U.urealyticuminfectionmaybein volvedinnon specificurethritis(NSU),prostati tis,postpartumfever,infertility,pelvicinflamma torydisease,neonatalpneumoniaandevenchronic lungdisease(CLD)[1].ItisknownthatU.urealyticumlackscellwallstructureandcontains abundantmembraneproteins,butitspathogenicity isstillunknownclearly.…     

大鼠大脑皮质微血管的一氧化氮合酶神经元及终末的正常分布研究

目的;研究正常大鼠大脑皮质额、顶、枕、颞叶的一氧化氮合酶（nitric oxide synthase,NOS）神经元及NOS阳性终末的分布,尤其注意它们与皮质微血管的关系。方法:采用NADPH-d组化方法。结果:NOS阳性神经元在皮质各层散在分布,数量较少,额、顶叶约半数的NOS神经元直接与皮质血管构成接触,枕、额叶中约三分之一的NOS神经元与皮质血管构成接触。而NOS阳性纤维多且密集成网状,额、顶叶的终末数量明显多于枕、颞叶,其中6％～7％的阳性终末分布至血管壁。结论:大脑皮质内的NOS神经元及NOS阳性终末参与皮质微循环的调节。     

Androgen receptor independent cardiovascular action of the antiandrogen flutamide

We have previously shown that flutamide (specific antagonist of the androgen receptor) has antihypertensive effects. In the present study we examined the mechanisms of flutamide action in the vasculature. The vascular effects of flutamide were assayed in aortae isolated from male or female Sprague-Dawley rats and from rats or mice lacking a functional androgen receptor ( tfm, testicular feminization mutation). The effect of flutamide on coronary flow was tested in isolated hearts. In addition, male hypertensive rats with tfm mutation were treated with flutamide, and blood pressure was monitored. Flutamide induced a relaxation of rat aortae from all the strains used (maximum relaxation at 10 microM: 51.3+/-5.2% of phenylephrine contraction) and increased the coronary flow. The aortic relaxation to flutamide was abolished by endothelium removal, or by inhibition of nitric oxide synthase, guanylyl cyclase, and tyrosine kinase but not by calmodulin inhibition. Flutamide treatment attenuated the development of hypertension in mouse renin transgenic rats with the tfm mutation. Flutamide produces direct vasodilation by inducing release of NO from the endothelium and causes subsequent activation of soluble guanylyl cyclase in an active androgen receptor independent manner. This response may contribute to the observed antihypertensive actions of flutamide.     

氧化型低密度脂蛋白对人脐静脉内皮细胞ICAM-1及NO表达的影响

目的研究氧化型低密度脂蛋白(oxidized low density liprotein,ox-LDL)对脐静脉内皮细胞细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)及一氧化氮(nitric oxide,NO)表达的影响.方法采用细胞ELISA法测定细胞表面ICAM-l的含量,硝酸还原酶法测定细胞培养上清液中NO,免疫组化法结合图象分析测定细胞一氧化氮合酶(nitric oxide synthase,NOS)含量.结果 ox-LDL可明显增加脐静脉内皮细胞ICAM-l的表达,并减少NO及NOS的表达,且有浓度依赖性,但无明显的时间依赖性.结论 ox-LDL可损害内皮细胞的功能,减少内皮细胞NO、NOS的表达并增加内皮细胞ICAM-1的表达.这可能为ox-LDL促进动脉粥样硬化的机制之一.     

细胞性一氧化氮供体的建立及其对肿瘤细胞凋亡的诱导效应

目的一氧化氮（ＮＯ）供体细胞系的建立及其对肿瘤细胞凋亡的诱导效应研究。方法采用３Ｈ－胍氨酸转换法进行ＮＯ合成酶（ＮＯＳ）活性测定；亚硝酸盐检测；ＤＮＡ片段的提取及凝胶电泳分析；凋亡细胞的ＤＡＰＩ染色观察；Ｗｅｓｔｅｒｎｂｌｏｔ分析。结果将ｉＮＯＳ真核表达质粒、ｐＣＭＶ／ｉＮＯＳ转染至Ｓｐ２／０骨髓瘤的变异株中，并获得能稳定表达ｉＮＯＳ和合成ＮＯ的重组细胞系（ＳＰｍｔ／ｉＮＯＳ），表达ｉＮＯＳ的效率为每升培养物含４００μｇ蛋白。利用该细胞作ＮＯ细胞性供体，成功地证实ＮＯ能诱导Ｓｐ２／０骨髓瘤细胞发生凋亡。结论所建细胞性ＮＯ供体应用于ＮＯ的生物学功能研究具有ＮＯ合成稳定；更能模拟体内细胞间ＮＯ的信息传递过程；不需任何外源刺激即可合成ＮＯ等独特优点。所建ＮＯ供体细胞可以用于肿瘤细胞凋亡的研究。