N,N'-diferuloyl-putrescine和JBIR-94的全合成

阿魏酸和1,4-丁二胺经缩合反应制得天然产物N,N-diferuloyl-putrescine(1);用Pd/C催化还原1首次完成了天然产物JBIR-94的全合成,总收率77.1%,其结构经1H NMR,13C NMR和IR确证。

Putrescine Plays a Positive Role in Salt-Tolerance Mechanisms by Reducing Oxidative Damage in Roots of Vegetable Soybean

Polyamines play important roles in plant tolerance to environmental stress. With the aim of investigating the possible involvement of putrescine （Put） in salt-tolerance mechanisms in vegetable soybean roots, exogenous Put （10 mmol L＂） and its biosynthetic inhibitor D-arginine （D-Arg） （0.5 mmol L-1） were added to nutrient solution when vegetable soybean （Glycine max L. cv. Huning 95-1） seedlings were exposed to 100 mmol L^-11 sodium chloride （NaCl）. The results showed that Put ameliorated but D-Arg aggravated the detrimental effects of NaCl on plant growth and biomass production. Under NaCl stress, levels of free, soluble conjugated and insoluble bound types of Put in roots of vegetable soybean were reduced, whereas those of free, soluble conjugated, and insoluble bound types of spermidine （Spd） and spermine （Spm） were increased. Exogenous Put eliminated the decrease in Put but promoted the increase of Spd and Spm. However, these changes could be reversed by D-Arg. Under NaCl stress, activities of arginine decarboxylase （ADC）, S-adenosylmethionine decarboxylase （SAMDC）, diamine oxidase （DAO）, and polyamine oxidase （PAO） were induced, with exogenous Put promoting and D-Arg reversing these changes. Furthermore, NaCl stress decreased activities of antioxidant enzymes. Exogenous Put alleviated but D-Arg exaggerated these effects of NaCl stress, resulting in the same changes in membrane damage and reactive oxygen species （ROS） production. These results indicated that Put plays a positive role in vegetable soybean roots by activating antioxidant enzymes and thereby attenuating oxidative damage.

Transepithelial transport of putrescine across monolayers of the human intestinal epithelial cell line, Caco-2

AIM To study the transepithelial transport characteristics of the polyamine putrescine in human intestinal Caco-2 cell monolayers to elucidate the mechanisms of the putrescine intestinal absorption.METHODS The transepithelial transport and the cellular accumulation of putrescine was measured using Caco 2 cell monolayers grown on permeable filters.RESULTS Transepithelial transport of putrescine in physiological concentrations (>0.5 mM)from the apical to basolateral side was linear. Intracellular accumulation of putrescine was higher in confluent than in fully differentiated Caco-2 cells, but still negligible (less than 0.5%) of the overall transport across the monolayers in apical-to-basolateral direction. EGF enhanced putrescine accumulation in Caco-2 cells by four-fold, as well as putrescine conversion to spermidine and spermine by enhancing the activity of Sadenosylmethionine decarboxylase. However,EGF did not have any significant influence on putrescine flux across the Caco-2 cell monolayers. Excretion of putrescine from Caco-2cells into the basolateral medium did not exceed 50 picomoles, while putrescine passive flux from the apical to the basolateral chamber,contributed hundreds of micromoles polyamines to the basolateral chamber.CONCLUSION Transepithelial transport of putrescine across Caco-2 cell monolayers occurs in passive diffusion, and is not influenced when epithelial cells are stimulated to proliferate by a potent mitogen such as EGF.

Putrescine mitigates intestinal atrophy through suppressing inflammatory response in weanling piglets

Background: Polyamines are essential for cell growth and beneficial for intestinal maturation. To evaluate the effects of putrescine on alleviating intestinal atrophy and underlying molecular mechanisms, both in vivo feeding trial and in vitro cell culture were conducted. Weanling pigs were fed a diet supplemented with 0, 0.1%, 0.2% or0.3% putrescine dihydrochloride, whereas porcine intestinal epithelial cells(IPEC-J2) were challenged with lipopolysaccharide(LPS) in the presence of 200 μmol/L putrescine.Results: Dietary supplementation with 0.2% putrescine dihydrochloride decreased the incidence of diarrhea with an improvement in intestinal integrity. Inhibition of ornithine decarboxylase activity decreased the proliferation and migration of IPEC-J2 cells, and this effect was alleviated by the supplementation with putrescine. The phosphorylation of extracellular signal regulated kinase and focal adhesion kinase was enhanced by putrescine. LPS increased the expression of inflammatory cytokines [tumor necrosis factor α(TNF-α), interleukin 6(IL-6) and IL-8],and inhibited cell proliferation and migration in IPEC-J2 cells. Adding exogenous putrescine suppressed the expression of TNF-α, IL-6 and IL-8, and recovered cell migration and proliferation in LPS-treated IPEC-J2 cells. Dietary putrescine supplementation also reduced the m RNA levels of TNF-α, IL-6 and IL-8 and their upstream regulator nuclear receptor kappa B p65 subunit in the jejunal mucosa of piglets.Conclusions: Dietary supplementation with putrescine mitigated mucosal atrophy in weanling piglets through improving anti-inflammatory function and suppressing inflammatory response. Our results have important implications for nutritional management of intestinal integrity and health in weanling piglets and other neonates.

Phosphatidylethanolamine N-methyltransferase and choline dehydrogenase gene polymorphisms are associated with human sperm concentration

Choline is a crucial factor in the regulation of sperm membrane structure and fluidity, and this nutrient plays an important role in the maturation and fertilizing capacity of spermatozoa. Transcripts of phosphatidylethanolamine N-methyltransferase （PEMT） and choline dehydrogenase （CHDH）, two basic enzymes of choline metabolism, have been observed in the human testis, demonstrating their gene expression in this tissue. In the present study, we explored the contribution of the PEMTand CHDHgene variants to sperm parameters. Two hundred oligospermic and 250 normozoospermic men were recruited. DNA was extracted from the spermatozoa, and the PEMT -774G〉C and CHDH ＋432G〉T polymorphisms were genotyped. The genotype distribution of the PEMT-774G〉C polymorphism did not differ between oligospermic and normozoospermic men. In contrast, in the case of the CHDH ＋432G〉T polymorphism, oligospermic men presented the CHDH432GIG genotype more frequently than normozoospermic men （62% vs. 42%, P〈0.001）. The PEMT774GIG genotype was associated with a higher sperm concentration compared to the PEMT774GIC and 774C/C genotypes in oligospermic men （12.5±5.6× 10^6 spermatozoa m1-1 vs. 8.3±5.2×10^6 spermatozoa m1-1, P〈0.002） and normozoospermic men （81.5±55.6×10^6 vs. 68.1±44.5×10^6 spermatozoa m1-1, P〈0.006）. In addition, the CHDH432G/G genotype was associated with higher sperm concentration compared to CHDH432G.T and 432T/T genotypes in oligospermic （11.8±5.1×10^6 vs. 7.8±5.3×10^6 spermatozoa m1-1, P〈0.003） and normozoospermic men （98.6±62.2×10^6 vs. 58.8±＋33.6×10^6 spermatozoa m1-1, P〉0.001）. In our series, the PEMT-774G〈C and CHDH ＋432G〈T polymorphisms were associated with sperm concentration. This finding suggests a possible influence of these genes on sperm quality.

Influence of Putrescine on Enzymes of Ammonium Assimilation in Maize Seedling

The effect of different concentrations of putrescine on biochemical changes in root and shoot of six days old maize seedlings in terms of enzymes of ammonium assimilation were examined. The results revealed that glutamate dehydrogenase (GDH) activity was enhanced at lower concentration of putrescine but at higher concentration, the activity of this enzyme was declined. Glutamine synthetase (GS) activity decreased with increase in concentration of putrescine and it was highest at 1000 μm concentration. Howe ver, glutamate synthase (GOGAT) activity increased with increase in concentration of putrescine upto 100 μm in root and upto 50 μm in shoot and further increase in concentration resulted in decline of enzymatic activity. Protein and total nitrogen content increased upto 10 μm concentration of putrescine and it decreased further with increase in concentration both in root and shoot of maize seedling.

Physiological and Proteomic Analyses Reveal Effects of Putrescine-Alleviated Aluminum Toxicity in Rice Roots

The effects of putrescine on improving rice growth under aluminum(Al)toxicity conditions have been previously demonstrated,however,the underlying mechanism remains unclear.In this study,treatment with 50 pmol/L Al significantly decreased rice root growth and whole rice dry weight,inhibited Ca2+uptake,decreased ATP syn thesis,and in creased Al,H2O2 and malon dialdehyde(MDA)con tents,whereas the application of putrescine mitigated these negative effects.Putrescine increased root growth and total dry weight of rice,reduced total Al content,decreased H2O2 and MDA contents by increasing antioxidant enzyme(superoxide dismutase,peroxidase,catalase and glutathione S・transferase)activities,increased Ca2+uptake and energy product!oru Proteomic analyses using data-independent acquisition successfully identified 7934 proteins,and 59 representative proteins exhibiting fold-change values higher than 1.5 were randomly selected.From the results of the proteomic and biochemical analyses,we found that putrescine significantly inhibited ethylene biosynthesis and phosphorus uptake in rice roots,increased pectin methylation,decreased pectin content and apoplastic Al deposit!on in rice roots.Putrescine also alleviated Al toxicity by repairing damaged DNA and increasing the proteins involved in maintaining plasma membra ne integrity and normal cell proliferation.These fin dings improve our understanding of how putrescine affects the rice response to Al toxicity,which will facilitate further studies on environmental protection,crop safety,in novations in rice performance and real-world producti on.

Putrescine Enhances Seed Germination Tolerance to Heat Stress in Arabidopsis thaliana

Putrescine(Put)as the compound of plant polyamines is catalyzed by arginine decarboxylase(ADC),which is encoded by two members,ADC1 and ADC2 in Arabidopsis,and ADC2 is mainly responsible for Put biosynthesis.Accumulated evidence demonstrates the important function of Put in plant growth and development,but its role in regulating seed germination under high temperature(HT)has not been reported yet.SOMNUS(SOM)is the negative regulator for seed germination thermoinhibition by altering downstream gibberellin(GA)and abscisic acid(ABA)metabolism.In this study,we found exogenous application of Put obviously alleviated the inhibition effect of HT on seed germination.Whereas pharmacological inhibition of endogenous Put level reduced seed germination under HT.Consistently,HT induced the rapid accumulation of Put level,and the adc2 mutant defi-ciency in Put biosynthesis also showed more sensitivity to HT stress.Furthermore,we found that the Put signal suppressed the expression of SOM and changed the transcriptional patterns of genes associated with GA/ABA metabolism.Genetic analysis also revealed SOM was epistatic to ADC2 to alter GA/ABA metabolism.Collectively,our finding reveals the novel function of Put in controlling seed germination under HT through SOM,and provides the possibility to develop Put as the innovational regulator for uniform seed germination under HT stress.

Effect of Putrescine on Low-Temperature Acclimation in Chlamydomonas reinhardtii

Putrescine is reported to be necessary for cold acclimation under low-temperature stress.In this study,the effect of low-temperature on some physiological and biochemical parameters has been investigated using the green algae Chlamydomonas reinhardtii.The lipid peroxidation rate,amount of Rubisco protein,activities of antioxidant enzymes and gene expression of polyamine biosynthesis(odc2,and spd1),heat shock proteins(hsp70c,hsp90a,and hsp90c),and PSII repair mechanisms(psba,rep27,and tba1)were determined to understand the low-temperature response.Exogenous putrescine application significantly increased Rubisco protein concentration and catalase enzyme activities under low-temperature stress.Moreover,real-time RT-PCR results and gene expression analysis showed that polyamine metabolism induced gene expression at low-temperatures in the first 24 h.In the same way,the gene expression of heat shock proteins(hsp70c,hsp90a,and hsp90c)decreased under low-temperature treatment for 72 h;however,application of putrescine enhanced the gene expression in the first 24 h.The results obtained indicated that molecular response in the first 24 h could be important for cold acclimation.The psba and tba1 expressions were reduced under low-temperatures depending on the exposure time.In contrast,the exogenous putrescine enhanced the expression level of the psba response to low-temperature at 24 and 72 h.The results obtained in this study indicate that putrescine could play a role in the PS II repair mechanisms under low-temperature stress.

多胺在肿瘤发生发展中的作用及其干预研究进展

多胺在肿瘤发生发展、靶向治疗和免疫治疗方面发挥着重要的作用。多胺包括腐胺、精胺和亚精胺等,临床研究和实验结果显示肿瘤存在多种形式的多胺代谢异常,且多胺代谢异常的研究对于肿瘤靶向治疗、免疫治疗生化标志物研究具有重要价值。随着对多胺在肿瘤发生发展和肿瘤治疗中作用的认识不断加深,人们对以多胺代谢为靶点的抗肿瘤干预策略的兴趣日益加深。本文综述多胺及其代谢与不同类型肿瘤进展的关系,旨在总结多胺代谢调控的药物靶点和临床防治药物开发策略。

探讨胃“炎癌转化”中腐胺与巨噬细胞极化的关系

浅表性胃炎逐步发展到胃癌的这个过程称之为Correa级联反应。在Correa级联反应中,炎症阶段的炎性微环境和肿瘤细胞所处肿瘤微环境都发现有巨噬细胞的参与。而多胺,尤其是腐胺,与胃癌的发生、发展密切相关,并且多胺的代谢可以影响巨噬细胞的极化。近年来,腐胺与胃“炎癌转化”的关系及通过调控巨噬细胞的极化来干预癌症的发生和发展已成为研究热点。本文综述了近年来涉及胃癌、腐胺和巨噬细胞极化的相关研究,探讨胃“炎癌转化”过程中腐胺与巨噬细胞极化的关系,期望后续研究能通过调节腐胺代谢和巨噬细胞极化来达到预防或延缓胃癌发生、发展的目的。

Cadaverine and putrescine exposure influence carbon and nitrogen cycling genes in water and sediment of the Yellow River

The response patterns of microbial functional genes involved in biogeochemical cycles to cadaver decay is a central topic of recent environmental sciences.However,the response mechanisms and pathways of the functional genes associated with the carbon(C)and nitrogen(N)cycling to cadaveric substances such as cadaverine and putrescine remain unclear.This study explored the variation of functional genes associated with C fixation,C degradation and N cycling and their influencing factors under cadaverine,putrescine and mixed treatments.Our results showed only putrescine significantly increased the alpha diversity of C fixation genes,while reducing the alpha diversity of N cycling genes in sediment.For the C cycling,the mixed treatment significantly decreased the total abundance of reductive acetyl-CoA pathway genes(i.e.,acsB and acsE)and lig gene linked to lignin degradation in water,while only significantly increasing the hydroxypropionate-hydroxybutylate cycle(i.e.,accA)gene abundance in sediment.For the N cycling,mixed treatment significantly decreased the abundance of the nitrification(i.e.,amoB),denitrification(i.e.,nirS3)genes in water and the assimilation pathway gene(i.e.,gdhA)in sediment.Environmental factors(i.e.,total carbon and total nitrogen)were all negatively associated with the genes of C and N cycling.Therefore,cadaverine and putrescine exposure may inhibit the pathway in C fixation and N cycling,while promoting C degradation.These findings can offer some new insight for the management of amine pollution caused by animal cadavers.

叶面喷施不同浓度腐胺复配剂对黄瓜幼苗高温抗性的影响

为缓解高温胁迫对设施黄瓜幼苗的伤害,以津春4号为试材,将腐胺(Put)、褪黑素(MT)、脯氨酸(Pro)和黄腐酸钾(MFA)以不同的浓度进行复配,研究不同浓度组合对黄瓜幼苗高温抗性、光合作用和抗氧化酶活性等的影响,筛选出缓解高温胁迫伤害最优的配方。结果表明:8 mmol·L^(-1)Put配合施用0.3 g·L^(-1)MFA、1.5 mmol·L^(-1)Pro及50μmol·L^(-1)MT能够有效缓解高温胁迫对黄瓜幼苗的伤害,采用隶属函数法综合评价效果最佳,与对照相比净光合速率增加了160.71%,H_(2)O_(2)含量降低了40.64%,可溶性蛋白含量增加了126.92%,SOD、POD、CAT活性分别提高了29.32%、31.66%、52.16%。

腐胺对枫桦和欧洲白桦嫩枝扦插生根及根茎养分含量的影响

【目的】枫桦和欧洲白桦作为用材林、土壤修复和城市绿化的优良树种,具有重要的经济和生态价值,优树嫩枝扦插是加快枫桦和欧洲白桦良种快速扩繁的重要途径,腐胺作为一类具有高生物活性的含氮碱与插穗生根关系密切,近年来受到诸多学者关注。研究腐胺对2个树种嫩枝扦插生根过程中的根系性状和根茎非结构化碳水化合物含量变化的影响,探讨其对2个树种嫩枝扦插生根作用效果的最佳浓度。【方法】以同家系枫桦和欧洲白桦2年生嫩枝为试验材料,设置不同浓度梯度T1、T2、T3(500、1000、1500 mg·L^(-1))的腐胺溶液浸泡组和对照组(CK1和CK2)处理,测定插穗不定根生根率、生根数、根长、根质量和根茎非结构化碳水化合物含量,通过方差分析、相关性分析和隶属函数分析揭示腐胺对2个树种插穗生根和养分含量的影响。【结果】腐胺对枫桦扦插生根促进效果最适浓度为500 mg·L^(-1),生根率达26.7%,生根数达5.3个,平均根长达9.4 cm;对欧洲白桦的最佳浓度为1000 mg·L^(-1),生根率达42.7%,生根数达6.7个,平均根长达6.5 cm。而对于根茎非结构化碳水化合物含量,1000 mg·L^(-1)浓度腐胺对枫桦根茎非结构化碳水化合物含量提升整体效果最佳,500 mg·L^(-1)浓度腐胺对欧洲白桦根茎非结构化碳水化合物含量的促进效果更好。【结论】腐胺浓度是影响2个树种扦插生根特性的关键因素,与传统处理方法ABT(生根粉)或IBA(吲哚丁酸)相比,腐胺不仅显著促进了2个树种插穗生根率、根长、根质量等性质,还对根茎非结构化碳水化合物含量具有提升作用,但通过隶属函数分析显示,腐胺+ABT(或IBA)整体指标促进效果虽然最优,但较仅使用ABT(或IBA)处理组该优势并不明显。

ART1 and putrescine contribute to rice aluminum resistance via OsMYB30 in cell wall modification

Cell wall is the first physical barrier to aluminum(Al)toxicity.Modification of cell wall properties to change its binding capacity to Al is one of the major strategies for plant Al resistance;nevertheless,how it is regulated in rice remains largely unknown.In this study,we show that exogenous application of putrescines(Put)could significantly restore the Al resistance of art1,a rice mutant lacking the central regulator Al RESISTANCE TRANSCRIPTION FACTOR 1(ART1),and reduce its Al accumulation particularly in the cell wall of root tips.Based on RNA-sequencing,yeast-onehybrid and electrophoresis mobility shift assays,we identified an R2R3 MYB transcription factor OsMYB30 as the novel target in both ART1-dependent and Put-promoted Al resistance.Furthermore,transient dual-luciferase assay showed that ART1 directly inhibited the expression of OsMYB30,and in turn repressed Os4CL5-dependent 4-coumaric acid accumulation,hence reducing the Al-binding capacity of cell wall and enhancing Al resistance.Additionally,Put repressed OsMYB30 expression by eliminating Alinduced H2O2accumulation,while exogenous H2O2promoted OsMYB30 expression.We concluded that ART1 confers Put-promoted Al resistance via repression of OsMYB30-regulated modification of cell wall properties in rice.

基于氧化香菇多糖金纳米簇复合物的电化学发光在腐胺检测中的应用

对香菇多糖进行氧化修饰,以氧化香菇多糖(OLNT)为模板,利用微波辅助加热法制备了氧化香菇多糖金纳米簇复合物(OLNT-AuNCs);利用腐胺对OLNT-AuNCs电化学发光(ECL)的猝灭效果,建立了腐胺传感器并用于3种市售葡萄酒中腐胺质量浓度的检测。结果表明:OLNT对AuNCs分散效果良好,AuNCs平均粒径为2.8 nm,晶格条纹间距为0.23 nm,在紫外光照射下,AuNCs可发出红色荧光,荧光峰值为612 nm;OLNT-AuNCs作为发光剂,在共反应剂K_(2)S_(2)O_(8)体系中具有良好的ECL特性,腐胺对该发光体系有稳定的特异性猝灭效果,ECL猝灭程度(ΔI_(ECL))与腐胺质量浓度对数(lg C)呈良好的线性相关关系(R^(2)=0.9975),线性范围为0.03~18.6 mg/L,检出限为9μg/L(S/N=3);3种市售葡萄酒样品中均检出腐胺,RSD均小于2%,平均加标回收率为91.2%~109.3%,该传感器重复性较好,回收率较高,可用于食品或生物样品中腐胺的检测。

多胺与幽门螺杆菌相关胃癌的研究进展

多胺的特点是分子质量小,具有一定的生物活性,是一类脂肪族化合物。多胺包括腐胺、精胺和亚精胺,是细胞生长和存活所必需的化合物。随着对多胺分子和细胞功能认识的进步,研究学者对多胺代谢在抗癌治疗中的作用越发感兴趣。本文论述了多胺的合成过程及胃癌发生过程中的多胺代谢。多胺与多种癌症有关。在这篇综述中,主要关注多胺失调在胃炎和随后的胃癌进展中的作用。由于多胺在癌症发展中的新意义,越来越多有前途的临床试验使用药物靶向多胺代谢途径来达到潜在的化学预防和抗癌治疗效果。

Nicotinamide N-methyltransferase and liver diseases

Cellular metabolism-induced epigenetic regulation is essential for the maintenance of cellular homeostasis.Nicotinamide N-methyltransferase(NNMT)is emerging as a key point of intersection between cellular metabolism and epigenetic regulation and has a central role in various physiological and pathological processes.NNMT catalyzes the methylation of nicotinamide(NAM)using the universal methyl donor S-adenosyl methionine(SAM)to yield S-adeno-syl-L-homocysteine(SAH)and N1-methylnicotinamide(MNAM),directly linking methylation balance with nicotinamide adenosine dinucleotide(NAD+)contents.NNMT acts on either the SAM-methylation balance or both NAD+metabolism,depending on the tissue involved or pathological settings where metabolic demand is increased.Under physiological conditions,the liver act as an essential metabolic organ with abundant NNMT expression,while NNMT hepatic function is not mediated by its methyltransferase activity due to other major methyltransferases such as glycine N-methyltransferase(GNMT)in the liver.However,hepatic NNMT,as well as its metabolite is improperly regulated and linked to the worse pathological states in liver diseases,including alcoholic liver disease,non-alcoholic fatty liver disease(NAFLD),liver cirrhosis,and hepatocellular carcinoma(HCC),suggesting a potential role in the process of liver diseases.In this review,we summarize how NNMT regulates cell methylation balance and NAD metabolism,and extensively outline the current knowledge concerning the functions of NNMT in hepatic metabolism including glucose,lipid and energy,with a specific focus on the contribution of NNMT to the pathophysiology of liver-related diseases.NNMT is involved in the development and progression of liver diseases.Understanding the complex NNMT regulatory network and its effects on pathogenesis could provide new therapeutic strategies in the context of liver diseases.

UPLC-MS/MS法同时测定中药注射液中5种生物胺的含量

目的 建立采用UPLC-MS/MS法同时测定中药注射液中组胺、腐胺、酪胺、色胺、亚精胺等5种生物胺含量的方法。方法 采用Waters ACQUITY UPLC BEH HILIC(100 mm×2.1 mm,1.7 μm)色谱柱,以乙腈-20 mmol/L甲酸铵(甲酸调pH为4.0)为流动相,梯度洗脱,流速0.3 ml/min,柱温40℃。在正离子多反应监测(MRM)模式下测定。结果 各组分在1~200 ng/ml浓度范围内线性关系良好(r>0.999),平均回收率(n=6)在93.5%~98.2%,RSD均在1.0%以内。结论 该方法操作简单,灵敏度高,重复性强,可以用于中药注射液中生物胺的检测。