自发性高血压大鼠心瓣膜上含神经肽Y和含降钙素基因相关肽神经的变化

用免疫组织化学（ＡＢＣ）方法，观察了含神经肽Ｙ（ＮＰＹ）、降钙素基因相关肽（ＣＧＲＤ）神经在自发性高血压大鼠（ＳＨＲ）心瓣膜分布的变化。结果表明，在ＳＨＲ二尖瓣、三尖瓣上含ＮＰＹ、含ＣＧＲＰ神经的密度，和对照Ｗｉｓｔａｒ－Ｋｙｏｔｏ（ＷＫＹ）大鼠相比较，无明显差异。但ＳＨＲ二尖瓣及三尖瓣含ＮＰＹ神经的Ｒ值，即心壁和乳头肌两个来源的含肽神经相延续所在的腱索数目与本片瓣膜上的总腱索数的比值，大于ＷＫＹ大鼠；而ＳＨＲ心瓣膜含ＣＧＲＰ神经的Ｒ值和ＷＫＹ大鼠无差异。本文对ＳＨＲ心瓣膜上含ＮＰＹ和含ＣＧＲＰ神经的变化作了讨论。     

维持性血透患者血小板提取液中NPY与NT的表达和意义

为研究慢性肾功能衰竭（ＣＲＦ）患者血液透析（ＨＤ）过程中血小板神经肽Ｙ（ＮＰＹ）与神经降压素（ＮＴ）的含量变化及其作用机制。从患者血闪中分离出血小板,采用放射免疫分析法,对血小板提取液和血浆中ＮＰＹ与ＮＴ含量进行ＨＤ前后的动态。结果显示：撮的血小板组比较,ＨＤ患者血小板提取液中ＮＰＹ含量明显降低,血浆中ＮＰＹ含量则明显增高,而血小板中提取液与血小板提取液中ＮＰＹ含量明显降低,血浆中ＮＰＹ含量则明显     

去甲肾上腺素促使血管平滑肌细胞内三磷酸肌醇含量的增加

研究去甲肾上腺素（ＮＥ）对自发性高血压大鼠（ＳＨＲ）的主动脉血管平滑肌细胞（ＶＳＭＣｓ）内三磷酸肌醇（ＩｎｓＰ３）含量的影响，并与对照组京都威斯特大鼠（ＷＫＹ）相比较，结果显示，在基础状态下ＳＨＲ和ＷＫＹ的ＶＳＭＣｓ内ＩｎｓＰ含量针明显差异，ＮＥ作用下ＷＫＹ的ＶＳＭＣｓ内ＩｎｓＰ３值迅速增加，１０秒钟同即达高峰，而ＳＨＲ的ＶＳＭＣｓ内ＩｎｓＰ３的增加呈双相性，除快相峰值明显大于ＷＫＹ外，慢相峰值也     

高血压冠状动脉平滑肌细胞的形态学变化

既往研究表明,高血压伴随有血管壁中层血管平滑肌细胞（ＶＳＭＣ）的肥大〔１,２〕。但是高血压未形成时冠状动脉中层ＶＳＭＣ是否已存在肥大及抗高血压药物对ＶＳＭＣ肥大的影响尚不清楚。本研究以自发性高血压大鼠（ＳＨＲ）和同种系正常血压的京都种大鼠（ＷＫＹ）为动物模型,通过观察ＳＨＲ高血压形成前后及抗高血压药物罗沙坦治疗后,左冠状动脉前降支中层ＶＳＭＣ的形态变化,了解高血压冠状动脉ＶＳＭＣ的形态学变化及抗高血压药物治疗对其的影响。１　材料和方法１１　实验动物及分组　雄性ＳＨＲ和ＷＫＹ由上海市高血压研究所…     

神经肽Y基因在哺乳动物细胞CHO中的稳定表达

以ＬｉｐｏｆｅｃｔｉｎＴＭ介导将ｐＲｃ／ＣＭＶＮＰＹ质粒转染入ＣＨＯ细胞，经Ｇ４１８筛选及Ｎｏｒｔｈｅｒｎｂｌｏｔ、ＥＬＩＳＡ和ＨＰＬＣ检测，得到稳定表达神经肽Ｙ（ＮｅｕｒｏｐｅｐｔｉｄｅＹ，ＮＰＹ）的细胞株，表达量为２～１０ｎｇ／（ｍｌ·１０７细胞）；表达时相分析结果显示转染细胞在转染后第４天表达量最高。     

缺血性脑血管病患者血浆和血小板提取液神经肽Y与神经降压素的改变

目的和方法：采用放射免疫分析法,观察５８ 例不同病因的缺血性脑血管病（ＩＣＶＤ） 患者血小板提取液和血浆中神经肽Ｙ（ＮＰＹ） 与神经降压素（ＮＴ） 的含量变化。结果：在ＩＣＶＤ 患者血小板提取液中ＮＰＹ 含量明显低于对照组,血浆中ＮＰＹ 则明显高于对照组。血小板中ＮＰＹ 减少与血浆浓度的增高呈明显负相关（ ｒ ＝ － ０ ．６０ , Ｐ＜ ０ ．０１） 。ＮＴ 含量在患者血小板液中显著增高。结论：ＮＰＹ 与ＮＴ 在ＩＣＶＤ 病理过程中,由血小板大量释放并积极参与了以缩血管效应为主的病理反应,是造成血栓形成、局部血管痉挛的重要病理因素     

神经肽Y基因在COS－7细胞中的表达

构建大鼠ＮＰＹ基因的瞬时表达载体ｐＳＶＬ－ＮＰＹ，以Ｌｉｐｏｆｅｃｔｉｎ~（ＴＭ）介导将其导入ＣＯＳ－７细胞中，Ｎｏｒｔｈｅｒｎｂｌｏｔ和ＨＰＬＣ检测结果显示：ｐＳＶＬ－ＮＰＹ质粒转染的ＣＯＳ－７细胞可表达ＮＰＹｍＲＮＡ及成熟肽ＮＰＹ。     

培养的大鼠血管平滑肌细胞血管紧张素Ⅱ受体的基因表达及调节

在培养的自发性高血压大鼠（ＳＨＲ）和正常血压ＷＫＹ大鼠的主动动脉平滑肌细胞（ＡＳＭＣ）模型，应用Ｎｏｒｔｈｅｒｎ杂交和逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）技术，分别检测ＡＳＭＣ中碱性成纤维细胞生长因子（ｈＦＧＦ）和血管紧张素Ⅱ（ＡＮＧⅡ）Ｉ型受体（ＡＴ１Ｒ）的基因表达。结果表明：ＳＨＲＡＳＭＣ中ｈＦＧＦ基因的基础表达和ＡＮＧⅡ刺激后的表达水平元旦明显高于ＷＫＹ大鼠；ｂＦＧＦ（１０ｎｍ／ｍｌ）对两种     

血液透析前后血小板提取液中NPY与NT含量的变化及意义

探讨血液透析（ＨＤ）过程对慢性肾功能衰竭患者（ＣＲＦ）血小板中神经肽Ｙ（ＮＰＹ）与神经降压素（ＮＴ）含量的影响及ＮＰＹ 与ＮＴ 的作用机制。我们采用特异性放射免疫分析方法,对血小板提取液和不含血小板血浆中ＮＰＹ 及ＮＴ 含量进行ＨＤ 前后的动态观察,并以３０ 例健康人作为对照。结果：正常对照组提取的血小板液中ＮＰＹ 与ＮＴ 含量分别为（５８１８±２１２９） ｎｇ／１０９ 和（２５３８±１３４３） ｎｇ／１０９。与对照组比较,ＨＤ 前ＣＲＦ患者血小板提取液中ＮＰＹ 含量明显降低,ＮＴ 显著高于对照组。而ＨＤ后ＣＲＦ患者血小板提取液中ＮＰＹ 与ＨＤ 前比较明显增高,ＮＴ 则明显降低。ＣＲＦ患者血浆ＮＰＹ 含量在ＨＤ 前明显高于对照组,ＨＤ后明显下降。且ＮＰＹ 与ＮＴ 之间具有明显的相关性。结论：在ＣＲＦ病理过程中,由血小板释放的ＮＰＹ 和ＮＴ 与５羟色胺等生物活性物质共同参与了以缩血管效应为主的免疫反应,是造成肾性高血压和肾血管痉挛的重要病理因素。     

血小板提取液NPY与NT在血液透析中的表达及意义

目的：探讨血液透析过程血小板中神经肽Ｙ与神经降压素的含量变化及意义。方法：ＨＤ患者３５例,对照组３０例,采用放射免疫分析法测定血小板提取液和血浆中ＮＰＹ及ＮＴ的含量。结果;与对照组比较,ＨＤ患者血小板提取液ＮＰＹ含量明显降低,血浆中ＮＰＹ含量明显增高,血小板提取液与血浆ＮＴ均显著升高。     

观察大鼠下丘脑NPY神经元的培养及瘦素对其mRNA表达的影响

目的观察大鼠下丘脑NPY神经元的培养及瘦素对其mRNA表达的影响。方法用无血清限定性培养基体外培养新生大鼠的下丘脑神经元,观察其生长状况;用NSE、NF、NPY抗血清检测神经元的鉴定和表达,并对三者阳性细胞的胞体和突起做统计学分析。给予Leptin10-10mol/L、10-8mol/L、10-6mol/L3个浓度,观察leptin对NPYm RNA表达的影响。结果培养的NSE和NF阳性细胞生长至第7天时均达高峰;NPY阳性神经元在培养7～10d时,亦处于一稳定时期。NPYmRNA的表达在给予Leptin10-10mol/L时,与对照组相比无显著性差异;10-8mol/L、10-6mol/L浓度时,表达增强,与对照组相比有显著性差异(P<0.05)。结论取材于新生大鼠的下丘脑亦可进行体外培养;培养一周时是研究单因素影响神经元的最佳实验时间;在离体环境下,高浓度的leptin(10-8mol/L、10-6mol/L)能促进NPYmRNA的表达,NPY可能表现为抑制GnRH分泌的作用。     

NPY and related substances

NPY exhibits a broad distribution throughout the body. NPY has been localized in neurons that synthesize norepinephrine or epinephrine and also in many cell bodies which are not catecholaminergic. Coexistence of NPY and several other peptides has also been observed. Accordingly, NPY displays a wide variety of functional activities depending on its location and coexistence with other substances, especially catecholamines. NPY exerts direct effects at several targets and also modulates the cellular response to catecholamines and to peptides such as LHRH. It is reasonable to expect that NPY will modulate the pre- and postjunctional effects of catecholamines at many of their targets in view of the distribution of NPY in central catecholaminergic neurons and throughout the preaortic and sympathetic chain ganglia. Virtually nothing is known at the time of this writing about NPY receptors and postreceptor transduction mechanisms at different sites of NPY activity. Equally mysterious are the pre- and postjunctional receptor-coupled transduction mechanisms which are involved in the modulation of catecholaminergic or other peptidergic effects by NPY. The distribution of NPY and its involvement in cardiovascular, GI, endocrine, and neuroendocrine systems suggest that NPY may be an extremely important regulator of a spectrum of physiological functions.     

NPY and NPY receptors in airway structural and inflammatory cells in allergic asthma

Purpose

Neuropeptide Y (NPY) level is elevated in allergic asthmatic airways and activation of NPY receptor-1 (NPY-Y1) on antigen‐presenting cells (APCs) is essential for T cell priming. Paradoxically, NPY-Y1 modulates hyper-responsiveness in T cells, suggesting a bimodal role for NPY in APCs and T cells. Therefore, determination of the temporal and spatial expression pattern of NPY and its receptors in asthmatic airways is essential to further understand the role of NPY in allergic asthma.

Methods

Lungs were isolated from control and acute and chronic stages of OVA-sensitized and challenged mice (OVA). Stains, including H&E, PAS, and trichrome, were used to determine the severity of lung pathology. The expression patterns of NPY and NPY-Y receptors in the airways were determined using ELISA and immunofluorescence. Cytokine levels in the BALF were also measured.

Results

NPY levels were undetectable in the BALF of control mice, but significantly increased in the OVA group at day 80. Levels of IL-4, TGF-β1 and TGF-β2, significantly increased and peaked on day 45 and decreased on day 80 in the OVA group, exhibiting an inverse correlation with NPY levels. NPY expression was localized to macrophage-like cells in the peri-bronchial and peri-vascular areas in the lung tissue. NPY-Y1 and -Y5 receptors were constitutively expressed by both structural and inflammatory cells in the lung tissue.

Conclusions

NPY produced by activated macrophage-like cells may be involved in regulating cytokine production and cellular activities of immune cells in asthma. However, it remains unclear whether such an increase in NPY is a defensive/compensatory mechanism to modulate the effects of inflammatory cytokines.     

双抗体夹心免疫PCR－检测神经肽Y

建立了一种双抗体夹心免疫ＰＣＲ方法。用神经肽Ｙ单克隆抗体包被；多克隆抗体作为夹心抗体；生物素标记的羊抗兔ＩｇＧ和游离的亲合素作为连接分子；生物素化的腺病毒六邻体基因重组质粒ＤＮＡ作为指示分子；用腺病毒六邻体基因的特异引物扩增指标分子。结果表明本实验方法检测神经肽Ｙ的敏感性可达５．０×１０~（－９）μｇ／ｍｌ，或５．０×１０~（－１０）ｇ／ｍｌ，比ＥＬＩＳＡ（０．７８μｇ／ｍｌ）敏感１．５６×１０~８或１．５６×１０~９倍。     

The intracellular mobility of NPY and a putative mitochondrial form of NPY in neuronal cells

Preproneuropeptide Y is a precursor peptide to mature neuropeptide Y (NPY), which is a universally expressed peptide in the central and peripheral nervous system. NPY is normally routed to endoplasmic reticulum and secretory vesicles in cells, which secrete NPY. In our previous studies, we found a functional Leucine7 to Proline7 (L7P) polymorphism in the signal peptide sequence of preproNPY. This polymorphism affects the secretion of NPY and causes multiple physiological effects in humans. The sequence of NPY mRNA contains two in frame kozak sequences that allow translation initiation to shift, and translation of two proteins. In addition to mature NPY_1–36 also a putative truncated NPY_17–36 with mitochondrial targeting signal is produced. The purpose of this study was to investigate the protein mobility of the putative mitochondrial fragment and the effect of the L7P polymorphism on the cellular level using GFP tagged constructs. The mobility was studied with fluorescence recovery after photobleaching technique in a neuronal cell line. We found that the mobility of the secretory vesicles with NPY_1–36 in cells with L7P genotype was increased in comparison to vesicle mobility in cells with the more abundant L7L genotype. The mobility in the cells with the putative mitochondrial construct was found to be very low. According to the results of the present study, the mitochondrial truncated peptide stays in the mitochondrion. It can be hypothesized that this could be one of the factors affecting energy balance of the membranes of the mitochondrion.     

Allelic variation in the NPY gene in 14 Indian populations

NPY is a 36-aminoacid peptide expressed in several areas of the nervous system. Neuropeptide Y (NPY) receptors represent a widely diffused system that is involved in the regulation of multiple biological functions. The human NPY gene is located in chromosome 7. The functional significance of coding Leu7Pro polymorphism in the signal peptide of preproNPY is known. Six hundred and fifty four individuals of 14 ethnic Indian populations were screened for three mutations in the NPY gene, including Leu7Pro. We found that the Pro7 frequencies among the studied populations were much higher than in previous studies from other parts of the world. The highest allele frequency of Pro7 was detected in the Kota population in the Nilgiri Hill region of south India, and this may reflect a founder event in the past or genetic drift. All populations followed the Hardy–Weinberg equilibrium for the assayed markers. A total of five haplotypes were observed, only two of which were found to occur with a high frequency in all populations. No linkage disequilibrium (LD) was observed across the tested alleles in any population with the exception of Leu7Pro and Ser50Ser in the Badaga population (χ ² = 13.969; p = 0.0001).