成人阴茎包皮及包皮系带内nNOS免疫阳性神经末梢的分布

目的：观察成人阴茎包皮与包皮系带内神经元型一氧化氮合酶（nNoS）免疫阳性神经末梢的分布。方法：免疫组织化学方法。结果：正常成人阴茎包皮及包皮系带内均有密集的nNOS免疫阳性神经末梢存在,这些神经末梢主要位于表皮基底层,呈树枝状或念珠状分布,大多成束走行。阴茎系带处nNOS免疫阳性神经末梢的分布密度明显大于阴茎包皮处。结论：一氧化氮（NO）参与了阴茎包皮及包皮系带感觉信息的传递。     

Expression of cyclooxygenase-2, alpha 1-acid-glycoprotein and inducible nitric oxide synthase in the developing lesions of murine leprosy

Murine leprosy is a chronic disease of the mouse, the most popular animal model used in biomedical investigation, which is caused by Mycobacterium lepraemurium (MLM) whose characteristic lesion is the macrophage-made granuloma. From onset to the end of the disease, the granuloma undergoes changes that gradually transform the environment into a more appropriate milieu for the growth of M. lepraemurium. The mechanisms that participate in the formation and maturation of the murine leprosy granulomas are not completely understood; however, microbial and host-factors are believed to participate in their formation. In this study, we analysed the role of various pro-inflammatory and anti-inflammatory proteins in granulomas of murine leprosy after 21 weeks of infection. We assessed the expression of cyclooxygenase-2 (COX-2), alpha acid-glycoprotein (AGP), and inducible nitric oxide synthase (iNOS) at sequential stages of infection. We also looked for the nitric-oxide nitrosylation product, nitrotyrosine (NT) in the granulomatous lesions of murine leprosy. We found that a pro-inflammatory environment predominates in the early granulomas while an anti-inflammatory environment predominates in late granulomas. No obvious signs of bacillary destruction were observed during the entire period of infection, but nitrosylation products and cell alterations were observed in granulomas in the advanced stages of disease. The change from a pro-inflammatory to an anti-inflammatory environment, which is probably driven by the bacillus itself, results in a more conducive environment for both bacillus replication and the disease progression.     

巨噬细胞诱导型一氧化氮合酶的表达调节机制

一氧化氮是一种重要的巨噬细胞免疫效应分子,它参与免疫调节和宿主防御反应.一氧化氮的生成主要由诱导型一氧化氮合酶调节,然而诱导型一氧化氮合酶表达的调节机制及信号通路尚不完全清楚.     

纳米银联合放疗对大鼠胶质瘤内iNOS表达和NO水平的影响

目的:研究局部使用纳米银联合放疗对大鼠胶质瘤内诱导型一氧化氮合酶(iNOS)表达和一氧化氮(NO)含量的影响。方法:将C6细胞接种于大鼠右侧尾状核,建模后第8 d,将荷瘤鼠随机分成放疗对照组和纳米银联合放疗组,分别立体定向注入等体积的去离子水及20μg的纳米银,并行单次电离辐射。观察放疗后瘤组织形态学变化,免疫组织化学法检测iNOS蛋白表达,硝酸还原酶法测定NO含量。结果:瘤组织放疗后细胞排列紊乱、密度减少。放疗后6 h胶质瘤内iNOS即有表达,随着时间的延长,iNOS表达进一步增强。放疗后6 h和24h,纳米银联合放疗组iNOS活性均较放疗对照组增高,差异有统计学意义(P<0.01或P<0.001);NO含量的变化与iNOS变化规律基本一致。结论:纳米银联合放疗能够增强iNOS表达,促进NO生成,瘤组织iNOS表达和NO含量改变可能在纳米银联合放疗杀伤胶质瘤细胞的过程中发挥一定的作用。     

慢性疲劳应激对大鼠伏隔核一氧化氮合酶表达的影响

目的:探讨长时间游泳引起的疲劳应激对大鼠伏隔核(NAc)中神经元型一氧化氮合酶(nNOS)表达的影响.方法:采用成年雄性大鼠连续4周每天强迫游泳至力竭,用免疫组织化学方法结合图像处理测定伏隔核中nNOS的表达.结果:长时间强迫游泳应激可使大鼠伏隔核巾nNOS免疫细胞化学反应阳性神经元的数量(106.7%)、面积(150.2%)和灰度值(11.3%)显著增加.结论:nNOS参与疲劳应激的形成,一氧化氮在伏隔核对应激调节中起重要作用.     

姜黄素对急性肺损伤大鼠肺组织诱导型一氧化氮合酶和内皮型一氧化氮合酶表达的影响

目的:探讨姜黄素对内毒素(LPS)致急性肺损伤(ALI)模型大鼠肺组织中诱导型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)表达的影响。方法:雄性SD大鼠采用随机数字表法分为对照组、急性肺损伤模型组及姜黄素治疗组。采用一次性气管内滴注内毒素4 mg/kg制备急性肺损伤模型大鼠,治疗组于造模前15 min腹腔注射姜黄素200 mg/kg,于造模后3、6、12 h及24 h取肺组织,观察并比较各组肺组织形态学改变,并检测肺湿/干重比、肺含水量和肺组织中NO的含量,real-time PCR及免疫印迹法检测iNOS和eNOS的mRNA和蛋白表达。结果:模型组大鼠肺湿/干重比、肺组织含水量及NO含量均较对照组明显升高,iNOS的mRNA和蛋白表达量较对照组显著上调,而eNOS的mRNA和蛋白表达量较对照组显著下调。与模型组相比,姜黄素治疗组肺湿/干重比、肺含水量、NO含量均明显降低,iNOS的mRNA和蛋白表达量明显下调,eNOS的mRNA和蛋白表达量明显上调。结论:姜黄素可下调内毒素致急性肺损伤模型大鼠肺组织iNOS表达,并匕调eNOS表达。     

一氧化氮在巨噬细胞的细胞毒效应中的作用

目的:探讨一氧化氮(NO)对巨噬细胞细胞毒效应的影响。方法:将20只昆明种小鼠分为两组,一组皮下接种S180横纹肌肉瘤细胞,另一组作为正常对照组。分别取两组小鼠腹腔灌洗液中的巨噬细胞与K562肿瘤细胞共同培养,检测在培养液中加入左旋精氨酸(L-Arg)和G-单甲基左旋精氨酸(L-NAME)后巨噬细胞杀伤率的改变。结果:正常小鼠腹腔灌洗液中的巨噬细胞在L-Arg和L-NAME存在的情况下细胞毒效应没有改变;荷瘤小鼠腹腔灌洗液中的巨噬细胞在L-Arg组和空白组中的细胞毒效应明显增强。结论:NO是巨噬细胞细胞毒作用的一个效应分子。     

姜黄素对IL-17诱导的人角质形成细胞NO合成以及iNOS表达的影响

目的:探讨姜黄素对IL-17诱导的人表皮角质形成细胞株(HaCaT细胞)NO合成以及诱导型一氧化氮合酶(iN-OS)的mRNA和蛋白表达的影响.方法:用IL-17刺激体外培养的HaCaT细胞,并分别加入3种浓度的姜黄素共培养24h.并收集细胞上清液、提取细胞总RNA、总蛋白,分别进行NO含量的测定、荧光定量PCR和Western blot实验,明确姜黄素对NO含量以及iNOS表达的影响.结果:IL-17能够有诱导HaCaT细胞NO以及iNOS的表达(P＜0.01).姜黄素有效下调NO合成量以及iNOS的mRNA(P ＜0.01)及蛋白表达(P＜0.01)水平.结论:姜黄素对IL-17诱导的HaCaT细胞NO分泌及iNOS的表达具有明显的抑制作用,从而为其对角质形成细胞相关的皮肤炎症性疾病的治疗提供了理论依据.     

去除窦弓神经引起下丘脑室旁核小细胞部氮能神经元持续激活(英文)

目的:旨在研究去除窦弓神经是否导致下丘脑室旁核(PVN)氮能神经元处于持续激活状态。方法:成年大鼠行窦弓神经去除术,1周后制备下丘脑脑片,进行还原型尼克酰胺腺嘌呤二核苷酸脱氢酶(NADPH-d)组织化学结合Fos免疫组织化学染色。结果:在PVN的内侧、背侧和外侧小细胞部有大量Fos阳性神经元分布,并与NADPH-d部分共存,但在PVN的室周和前小细胞部以及大细胞仅观察到弱阳性的Fos信号,偶尔观察到双标记神经元。结论:去窦弓神经大鼠下丘脑室旁核小细胞部氮能神经元处于持续激活状态,可能起代偿性抑制中枢交感活性的作用。     

Expression of inducible nitric oxide synthase and nitrotyrosineduring the evolution of experimental pulmonary tuberculosis

Nitric oxide (NO) is a relevant antimycobacterial factor in mouse macrophages. NO is a product of inducible nitric oxide synthase (iNOS). NO toxicity is greatly enhanced by reacting with superoxide to form peroxynitrite that reacts with many biological molecules. Tyrosine is one of the molecules with which NO reacts and the product is nitrotyrosine (NT). The production of peroxynitrite and the nitrosylation of proteins might play a role in bacterial killing and also in mediating host injury. In this study, we used a well-characterized mouse model of pulmonary tuberculosis to examine the local kinetics of expression and cellular distribution of iNOS and NT at the cellular and subcellular level. The histopathological study showed two phases of the disease: early and late. The early phase was characterized by mononuclear inflammation and granuloma formation. During this phase, high percentages of activated macrophages were observed that were immunostained for iNOS and NT. Immuno-electronmicroscopy showed NT immunoreactivity in lysosomes and mycobacterial wall and cytoplasm. The concentration of iNOS mRNA and NO metabolites were also elevated. The late phase was characterized by progressive pneumonia with focal necrosis and a decrease of iNOS mRNA and NO metabolites. The strongest NT immunostained areas were the necrotic tissue. Macrophages became foamy cells with scarce iNOS immunostaining but strong NT immunoreactivity. At the ultrastructural level, these cells showed NT immunolabeling in cytoskeleton, mitochondria, lysosomes and cell membrane. NT was also located in bronchial epithelial cell mitochondria, in cell membranes and cytoplasm of endothelial cells and in actin bundles within smooth muscle cells. These results suggest an important role of NO in mycobacterial killing, particularly during the early phase of the infection. They also suggest an important participation by NO in tissue damage during the late phase of the disease.     

缺氧诱导因子-1在缺氧诱导一氧化氮合成酶基因表达中的作用

目的 研究缺氧诱导因子-1(HIF-1)在一氧化氮合成酶基因缺氧诱导反应中的作用。方法 体外合成具有HIF-1特异结合位点的DNA片段(红细胞生成素3'-增强子片段),借助脂质体,转入培养的鼠主动脉内皮细胞和肺微血管细胞,用半定量RT-PCR方法测定诱导型一氧化氮合成酶(iNOS)mRNA。结果 (1)大鼠主动脉内皮细胞、肺微血管内皮细胞在常氧下培养,有iNOS基因表达;(2)缺氧能诱导这两种细胞iNOS基因表达增加;(3)野生型EPO3'-增强子片段能阻断缺氧对内皮细胞iNOS基因表达的诱导作用,而突变片段则无此作用。结论 在iNOS基因序列中,可能存在EPO3'-端增强子片段,其参与内皮细胞的缺氧反应。     

NADPH-diaphorase activity and nitric oxide synthase isoforms in the trophoblast of Calomys callosus

The pattern of expression of a variety of placental nitric oxide synthase isoforms has contributed to elucidating the regulatory mechanisms of nitric oxide (NO) synthesis during gestation. The maintenance of vascular tone, attenuation of vasoconstriction, prevention of platelet and leukocyte adhesion to the trophoblast surface, and possible participation in uterine blood flow seem to be the main functions of NO generated at the fetal-maternal interface in humans and mice. Extending this knowledge to other rodent species commonly used as laboratory animals, in this study we focus on NADPH-diaphorase activity and the distribution of nitric oxide synthase isoforms (NOS) in the trophoblast cells of Calomys callosus during different phases of pregnancy. NADPH-diaphorase activity was evaluated cytochemically and the presence of NOS isoforms detected by immunohistochemistry. These techniques were performed on pre- and postimplantation embryos in situ and in vitro, as well as in placentae on d 14 and 18 of pregnancy. Neither NADPH-diaphorase activity nor inducible or endothelial NOS isoforms were found in pre-implanting embryos except after culturing for at least 48 h, when some of the embryonic cells were positive for the diaphorase reaction. On d 6·5 of pregnancy, trophoblast cells showed intense diaphorase activity both in situ and under in vitro conditions. A positive reaction was also found in the different placental trophoblast cells on d 14 and 18 of pregnancy. The inducible NOS (iNOS) isoform, but not the endothelial isoform, was immunodetected in trophoblast cells from the placenta and from postimplantation embryos in situ and under in vitro conditions. These results strongly suggest the production of NO by the iNOS isoform in the trophoblast of Calomys callosus after embryo implantation. The data also emphasise a possible role for the trophoblast in producing and releasing cytotoxic molecules at the fetal-maternal interface.     

诱导型一氧化氮合酶在糖尿病性阴茎勃起功能障碍大鼠阴茎的表达

目的:探讨诱导型一氧化氮合酶(iNOS)在糖尿病性阴茎勃起功能障碍(ED)发病进程中的可能作用.方法:注射链脲佐菌素建立糖尿病大鼠模型,分别在注射8周和12周后观察阴茎勃起次数,取大鼠阴茎,用ABC免疫组织化学方法检测阴茎iNOS的分布与表达,免疫印迹检测阴茎iNOS蛋白量的变化.结果:糖尿病大鼠的阴茎勃起次数低于对照组,并随病程延长降低;与对照组比较,糖尿病组阴茎内iNOS阳性细胞数和平均光密度、iNOS蛋白量升高,并随病程延长而进行性升高.结论:糖尿病组大鼠阴茎iNOS表达的升高与ED相关.     

Distribution of Nitrergic Neurons in the Dorsal Root Ganglia of the Bottlenose Dolphin (Tursiops truncatus)

Dorsal root ganglia (DRGs) contain the cell bodies of primary afferent neurons that transmit sensory information from the periphery into the spinal cord. Distinct populations of DRG neurons have been characterized by a variety of different immunohistochemical markers. A subpopulation of ganglionic neurons containing neuronal nitric oxide synthase (nNOS), an enzyme known to generate nitric oxide, has been detected in a number of mammalian species. Despite previous studies, no information is known on the presence and exact distribution of nNOS‐immunoreactive neurons in the DRGs of the bottlenose dolphin. In this investigation, immunoperoxidase for nNOS was used to determine the distribution and the perikaryal size of nitrergic neurons in the DRGs of this species. Double immunofluorescence protocol was used to determine the percentage of nNOS‐immunoreactive (IR) neurons over the total primary afferent neurons. In addition, double immunostaining was used to verify whether there was colocalization of nNOS with substance P (SP). In all DRGs, a subpopulation of small‐ and medium‐sized neurons (about 9%) exhibited nNOS immunoreactivity. Data analysis revealed that the majority of nNOS‐IR neurons (81.3%) expressed SP. The density of nNOS‐immunoreactive and nNOS/SP‐double immunopositive cells was relatively constant throughout the ganglia. However, as observed in others mammals, the number of nitrergic neurons decreased in the caudalmost DRGs. Our results, in conjunction with previous observations, suggest that nNOS‐IR neurons may be involved in the afferent transmission of visceral and nociceptive information as well as in the regulation of the vascular tone. Anat Rec,, 2011. © 2011 Wiley‐Liss, Inc.     

糖尿病大鼠视网膜神经源性一氧化氮合酶表达及乙酰胆碱酯酶的变化

目的:观察糖尿病大鼠视网膜神经源性一氧化氮合酶(nNOS)蛋白和基因水平的表达以及乙酰胆碱酯酶(AchE)的变化。方法:注射链脲佐菌素(STZ)建立糖尿病大鼠模型,于注射后12周和16周时将模型组及对照组大鼠的眼球冰冻切片。用原位杂交法、免疫组织化学法和组织化学法分别显示nNOS mRNA、nNOS和AchE阳性神经元或神经纤维,RS IMAGE软件进行图像分析处理。结果:与对照组比较,糖尿病大鼠视网膜nNOS mRNA、nNOS、AchE的光密度值均降低,12、16周都有显著性差异。结论:糖尿病大鼠视网膜nNOS基因转录和表达均下降,AchE含量降低,导致了NO水平下降,成为糖尿病性视网膜病变的主要因素之一。