植物源食品多胺的作用及其含量调节

多胺(polyamines, PAs)在植物源食品中广泛分布,在人体健康中发挥着重要作用。简要概述了多胺在细胞增殖、肿瘤发展中的作用及其作用的初步机理。通过化学调控和基因工程手段以及一些环境条件的控制,可以人为地调节植物源食品多胺的含量。     

多胺及其在植物体内的作用

1679年,勒力荷克观察到动物的精液中存在着一种星形和针伏结晶物质,1924年该结晶被证明是精胺磷酸盐。以后,逐渐确定了这类物质的化学结构和性质。然而长期以来。多胺一直被认为是无用的代谢末端产物和废物。从本世纪60年代以来,美国耶鲁大学加尔斯通(Galston)实验室对多胺作了系统地研究,认识到多胺具有刺激生长和防止衰老等作用,并开始联系到农业生产实践。最先提出多胺可能是一类新的植物激素,但后来更多的人认为可能是似cAMP那样的“第二信使”,以调节植物的生长和发育。本文就植物体中多胺的结构、分布、生物合成与氧化及其生理作用作一介绍。     

多胺调控细胞生长机制的研究进展

多胺(腐胺、精脒、精胺)是真核细胞体内重要的多聚阳离子,对细胞的生长增殖发挥重要作用。快速生长的细胞内含有高浓度的多胺,降低多胺含量将抑制细胞的生长,阻滞细胞周期,而升高多胺含量则会促进细胞快速生长增殖。对多胺的调控已经成为研究细胞生长增殖调控的切入点之一。     

多胺在植物生长发育过程中的生理作用

多胺在植物生长发育过程中具有广泛的生理作用,如参与植物衰老进程的调控、体细胞胚发生、花芽分化、花和果 实的发育及参与各种生理胁迫反应等。本文重点综述了多胺在植物生长发育过程中生理学功能方面的研究进展,并对有关 问题进行了讨论和展望。     

部分植物中的多胺氧化酶活性

多胺氧化酶（PAO）已证实在动物、微生物和植物中都有存在。目前,植物中的研究仅限于豆科和禾本科及百合、菊芋等为数不多的植物种类。本文研究了PAO在一些植物中的分布情况,取材涉及16个科28种植物。豌豆、直豆、蚕豆、花生、小麦等播种出苗后沙培（后期淋施Hoagland溶液）,满江红为人工养殖,其它材料均为新鲜时采集。大部分植物取用幼嫩组织（尤其是幼叶）,两个萍类植物用全植株测定。PAO的提取参照＊Mu＊＊。ri等［Ph～k加”,正984,23（2）：241）的方法,活性测定参照王富民等的方法「植物生理学报,1988,豆毛（4）。35叼…     

植物的多胺氧化酶

介绍了多胺氧化酶在植物中的分布及其反应特性、结构和成分、细胞和亚细胞定位、生理功能等方面的研究概况。     

植物体内微量多胺的分离和测定

目前对植物组织内多胺的含量主要是将植物材料的直接提取液和苯甲酰氯或和丹磺酰氯反应后,利用高效液相色谱仪测定。此方法在测定多胺含量较高的胚、芽和幼果时尚可,但对含量较少的材料如开花后一个月的果实,则无法测定。本文采用阳离子交换树脂来分离和浓缩多胺。对Flores和Galston的方法作了改进,用以测定梨果实中微量的多胺,减少了干扰,提高了灵敏度。     

多胺与植物衰老关系研究进展

多胺作为生理活性物质与植物衰老关系密切。本文综述了近十年来多胺对衰老的调控作用,从调节细胞膜的理化性质,生物大分子合成作用以及多胺与乙烯的关系等方面阐述了多胺延缓衰老的机制,比较了多胺和影响衰老的植物激素在信号转导过程中的作用。     

检测植物组织中多胺含量的高效液相色谱法

对反相高效液相色谱技术测定苯甲酰化多胺的方法进行了探讨 ,确定和研究了苯甲酰化反应的最佳温度、时间和影响苯甲酰化多胺稳定性的因素 ,优化了多胺的色谱分析条件     

Cyclin D1与细胞周期调控

细胞周期是细胞生命活动中一个最重要的过程,其关键是G1 期的启动.细胞周期蛋白(Cyclin)、细胞周期蛋白依赖性激酶(CDKs)和CDK抑制因子(CKIs)是参与钿胞周期调控的主要因子.Cyclin D1是调控细胞周期G1期的关键蛋白,是一个比其他Cyclins更加敏感的指标,对细胞周期调控至关重要.综述Cyclin D1的结构和功能及其在肿瘤组织中的表达特征,初步分析Cyclin D在昆虫细胞周期调控的研究.     

Polyamine Regulation of the Microtubule-Associated Protein Kinase

Microtubule protein prepared by cycles of assembly-disassembly contains a cyclic AMP-dependent protein kinase that phosphorylates the high-molecular-weight microtubule-associated protein MAP-2. The polyamine spermine at 2mM affected the phosphorylation of MAP-2 in a manner that depended on the cyclic AMP concentration. At cyclic AMP concentrations below 10(-6) M, spermine increased the rate of phosphorylation, while at cyclic AMP concentrations above 10(-6) M, spermine decreased the rate of phosphorylation. Spermine also decreased the final extent of cyclic AMP-dependent phosphorylation but did not affect the protein substrate specificity of the microtubule-associated protein kinase. MAP-2 was the principal substrate both in the presence and in the absence of spermine. Because of these results, we propose that microtubule protein phosphorylation may be regulated in vivo by spermine as well as by cyclic AMP levels.     

Ribosome Modulation Factor,an Important Protein for Cell Viability Encoded by the Polyamine Modulon

We searched for proteins whose synthesis is enhanced by polyamines at the stationary phase of cell growth using an Escherichia coli polyamine-requiring mutant in which cell viability is greatly decreased by polyamine deficiency. The synthesis of ribosome modulation factor (RMF) was strongly enhanced by polyamines at the level of translation at the stationary phase of cell growth. In rmf mRNA, a Shine-Dalgarno (SD) sequence is located 11 nucleotides upstream of the initiation codon AUG. When the SD sequence was moved to the more common position 8 nucleotides upstream of the initiation codon, the degree of polyamine stimulation was reduced, although the level of RMF synthesis was markedly increased. Polyamine stimulation of RMF synthesis was found to be caused by a selective structural change of the bulged-out region of the initiation site of rmf mRNA. The decrease in cell viability caused by polyamine deficiency was prevented by the addition of a modified rmf gene whose synthesis is not influenced by polyamines. The results indicate that polyamines enhance cell viability of E. coli at least in part by enhancing RMF synthesis.     

外源亚精胺对盐胁迫下黄瓜幼苗游离态多胺含量和多胺合成酶基因表达的影响

以黄瓜品种‘津春2号’（Cucumis sativusL.cv.Jinchun No.2）为材料,采用营养液栽培,研究了外源亚精胺（Spd）对NaCl胁迫下黄瓜幼苗叶片游离态多胺含量和多胺合成酶基因表达的影响。结果表明,75 mmol/LNaCl胁迫下,幼苗株高、茎粗和干鲜重显著降低,外源喷施1 mmol/L Spd处理可明显缓解盐胁迫对幼苗生长的抑制。盐胁迫下叶片游离态多胺含量显著增加,外源Spd进一步促进了游离态Spd和精胺（Spm）的积累,降低了游离态腐胺（Put）的积累。多胺合成酶基因表达分析表明,盐胁迫上调了adc、odc、samdc和spds基因的表达,施用外源Spd后进一步上调了samdc基因,下调了adc、odc、spds基因的表达。表明外源Spd参与了黄瓜幼苗体内多胺代谢的调节,通过下调盐胁迫下adc、odc基因的表达,抑制游离态Put的积累,上调samdc基因的表达促进游离态Spd和Spm的积累,进而缓解盐胁迫对植物生长的抑制。     

多胺生物合成的抑制对转化细胞c－Ha－ras癌基因表达的调控

抗多胺代谢剂──二氟甲基鸟氨酸（ＤＦＭＯ）作用于经含点突变的Ｈａ－ｒａｓ基因片段转染的转化细胞（ＨＲ－１细胞）引起细胞生长的抑制,其抑制率随ＤＦＭＯ浓度的增加而增大,此时细胞多停滞于Ｇ_１期;多胺合成的关键酶鸟氨酸脱羧酶（ＯＤＣ）活性显著下降;Ｈａ－ｒａｓ癌基因ｍＲＮＡ及ｒａｓＰ~（２１）蛋白的表达受到抑制;而外源性腐胺与ＤＦＭＯ的同时加入可防止上述一系列改变的发生,说明ＤＦＭＯ使ＨＲ－１细胞某些表型向亲本细胞逆转的作用是与细胞多胺生物合成的抑制直接相关。     

Effects of MDL 72527, a Specific Inhibitor of Polyamine Oxidase, on Brain Edema, Ischemic Injury Volume, and Tissue Polyamine Levels in Rats After Temporary Middle Cerebral After Occlusion

Abstract The possible effects of the polyamine interconversion pathway on tissue polyamine levels, brain edema formation, and ischemic injury volume were studied by using a selective irreversible inhibitor, MDL 72527, of the interconversion pathway enzyme, polyamine oxidase. In an intraluminal suture occlusion model of middle coerebral artery in spontaneously hypertensive rats, 100 mg/kg MDL 72527 changed the brain edema formation from 85.7 ± 0.3 to 84.5 ± 0.9% in cortex ( P < 0.05) and from 79.9 ± 1.7 to 78.4 ± 2.0% in subcortex (difference not significant). Ischemic injury volume was reduced by 22% in the cortex ( P < 0.05) and 17% in the subcortex ( P < 0.05) after inhibition of polyamine oxidase by MDL 72527. There was an increase in tissue putrescine levels together with a decrease in spermine and spermidine levels at the ischemic site compared with the nonischemic site compared with the nonischemic site after ischemia-reperfusion injury. The increase in putrescine levels at the ischemic cortical and subcortical region was reduced by a mean of 45% with MDL 72527 treatment. These results suggest that the polyamine interconversion pathway has an important role in the postischemic increase ini putrescine levels and that blocking of this pathway can be neuroprotective against neuronal cell damage after temporary focal cerebral ischemia.     

Roles and modes of action of nectins in cell–cell adhesion

Nectins are Ca²⁺-independent immunoglobulin (Ig)-like cell–cell adhesion molecules (CAMs), which comprise a family consisting of four members. Each nectin homophilically and heterophilically trans-interacts and causes cell–cell adhesion. Biochemical, cell biological, and knockout mice studies have revealed that nectins play important roles in formation of many types of cell–cell junctions and cell–cell contacts, including cadherin-based adherens junctions (AJs) and synapses. Mode of action of nectins in the formation of AJs has extensively been investigated. Nectins form initial cell–cell adhesion and recruit E-cadherin to the nectin-based cell–cell adhesion sites. In addition, nectins induce activation of Cdc42 and Rac small G proteins, which eventually enhances the formation of cadherin-based AJs through the reorganization of the actin cytoskeleton. Nectins furthermore heterophilically trans-interact with nectin-like molecules (Necls), other Ig-like CAMs, and assist or modify their various functions, such as cell adhesion, migration, and proliferation. We describe here the roles and modes of action of nectins as CAMs.     

Polyamine contents and amino acid decarboxylation activities of extremely halophilic archaebacteria and some eubacteria

An extremely halophilic archaebacterium Halobacterium cutirubrum was demonstrated to be devoid of any polyamine except agmatine when grown in a synthetic medium with no exogenous polyamines. Decarboxylation activities of homoarginine and canavanine as well as of arginine were shown to be present in cell lysates of 5 strains of extreme halophiles examined. H. halobium R1 was shown to have an additional pathway to synthesize agmatine from glutamic acid.     

植物器官大小相关基因研究进展

器官大小是植物形态的一个重要特征,而且具有严格的种属特异性。植物器官大小虽然受到外在的环境因素（如光照、营养等）的影响,但它由内在特有的细胞数目和细胞大小决定。许多通过转录调节、蛋白合成、激素调节或松弛细胞壁等途径作用于植物细胞繁殖和/或细胞扩张的基因已经被鉴定,它们的过表达或缺失表达能促进植物器官大小和加快植物生长。尽管如此,这些基因通过相对独立的途径起作用,在植物中难以阐明一个相对整合的器官大小基因调控网络,这也是该研究领域的亟待需要解决的问题。目前,一些器官大小相关基因已经应用农作物育种,并培育出显著增大的农作物品种,这也证实了利用器官大小基因进行植物品种选育的可行性。因此,通过研究药用植物器官大小的基因,人为地在分子水平上有目的的调控器官的大小和形态,是缓解当前许多药用植物面临的资源紧缺、枯竭濒危困境的可考虑途径之一。     

Regulation of Ganglioside Metabolism by Phosphorylation and Dephosphorylation

Abstract: Cell differentiation is frequently accompanied by alterations in the composition of gangliosides in the plasma membrane resulting from a regulation of the enzyme activities involved. The regulation of CMP-NeuAc:GM1 α2-3-sialyltransferase (ST-IV) and UDP-GalNAc:GM3 N-acetylgalactosaminyltransferase (Gal-NAc-T) by the degree of enzyme phosphorylation was analyzed by determination of the enzyme activity on incubation of NG108-15 cells with various protein phosphatase inhibitors (okadaic acid and orthovanadate) or protein kinase activators (phorbol ester and forskolin). Incubation with okadaic acid, but not with orthovanadate, inhibited the ST-IV activity to 45% of that of control cells with t_1/2 = 60 min for the inactivation reaction. This indicates a rapid hyperphosphorylation of ST-IV due to the inhibition of a serine/threonine-specific phosphatase. A similar rate of inactivation was found on stimulation of protein kinase C with phorbol ester. In contrast to ST-IV, the activity of GalNAc-T was increased on stimulation of intracellular phosphorylation systems. The fastest activation of GalNAc-T was achieved with forskolin, yielding up to 160% of the initial activity within 30 min of effector incubation. Up-regulation of GalNAc-T in conjunction with down-regulation of ST-IV by stimulation of phosphorylation is suggested to serve as a physiological mechanism to increase the concentration of GM1, which was found to be elevated in correlation with the cell density. This assumption was corroborated by metabolic labeling studies with radioactive ganglioside precursors indicating an enhancement of the relative amount of a-series gangliosides subsequent to GM3 on phosphorylation stimulation. In particular, the biosynthesis of GM1 was specifically elevated within 2 h of incubation with forskolin. We conclude from the overall data that the ganglioside composition during the cell differentiation of NG108-15 cells can be specifically regulated by both protein kinase A- and protein kinase C-related phosphorylation systems.