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The aim of this study was to contrast the effects of drought stress on polyamine oxidases gene expression andactivity as well as photosynthetic efficiency in relatively tolerant (Karoon) and sensitive (260) maize genotype.d Reduction in leaf relative water content as a result of drought led to increase in root growth, butdiminished shoot growth indices. Under drought stress, activity of antioxidant enzyme, catalase, significantlyincreased in both genotypes, whereas significant higher activity of superoxide dismutase and peroxidase wasonly observed in Karoon genotype. Expression of polyamine oxidase (PAO) genes (zmPAO1, zmPAO2,zmPAO3, zmPAO4, zmPAO5, zmPAO6) and activity of enzymatic polyamine oxidation was increased in bothgenotypes under drought stress. The enhancement in PAO gene expression and enzyme activity was moreprominent in Karoon cultivar compared to 260. Chlorophyll a fluorescence and fast induction kinetics werenegatively influenced by drought stress. These parameters were more affected in 260 cultivar compared withKaroon. Our results suggest that under drought stress, higher activity of polyamine oxidase pathway in backconversion of Spermine and spermidine to putrescine (protectant of photosynthetic apparatus) as well as higherantioxidant enzymes activity in Karoon cultivar, may play a role in higher efficiency of photosynthetic processin this cultivar
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The purpose of this study is to further explore the effects of SI-4650, a newly discovered small molecule inhibitor of spermine oxidase (SMO) in our laboratory, on proliferation and migration of human osteosarcoma 143B cells and its underlying molecular mechanism. Chemiluminescence and high performance liquid chromatograph were used to analyze the effect of SI-4650 on SMO activity in 143B cells. DCFH-DA-staining/FCM was used to analyze the accumulation of cellular reactive oxygen species (ROS), whereas MTT and FCM were used to detect proliferation and cell cycle. Transwell culture and Western blot were used to analyze the expression levels of migration-related proteins. PI/FITC-Annexin V/FCM, fluorescence microscopy and Western blot were used to analyze apoptosis and autophagy. Our results showed that SI-4650 could significantly decrease SMO activity, inhibit cell proliferation or migration, and induce a S-phase cell cycle arrest in 143B human osteosarcoma cells. The mechanism may be related to interfering with polyamine metabolism, activating mitochondrial-mediated apoptosis and causing autophagic death. These results suggest that SI-4650 has the potential for clinical use in treatment of osteosarcoma.
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Aim To investigate the effects of a novel spermine oxidase (SMO) inhibitor SI-4650 on proliferation, apoptosis and autophagy of human glioma U87MG cells as well as its possible mechanism. Methods MIT assay was used to detect cell proliferation. The chemiluminescence assay was used to determine enzyme activities of SMO and N'-acetylpolyamine oxidase (APAO). High performance liquid chromatography (HPI.C) was performed to detect cellular polyamine content. Transwell assay was used to evaluate the migration ability of U87MG cells. PI single-staining/flow cytometry( FCM ) were used to analyze cell cycle. PI/ FITC-Annexin V double-staining/ FCM and Western blot were used to analyze apoptosis. Confocal laser scanning microscopy ( LSCM) and Western blot were used to analyze autophagy. Results SI-4650 could significantly inhibit the activities of SMO and APAO, interfere with polyamine metabolism and reduce the content of total polyamine in U87MG cells. Treatment of U87MG cells with SI-4650 inhibited the proliferation and migration, caused G0/G, cell cycle arrest and induced apoptotic and autophagic cell death. Conclusions SI-4650 has the pharmacological activity of killing glioma U87MG cells, and its mechanism may be related to the interference of polyamine metabolism and induction of cell apoptosis and autophagy.
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Aim To discuss the changes of polyamine metabolism and autophagy in aging rat heart and the effect of exogenous spermidine on autophay in aged rat heart. Methods Western blot assay was used to detect the expressions of ODC and SSAT-rate-limiting enzyme of polyamine anabolism and catabolism as well as the expression of autophagy-relevant protein LC3?/? and p62 in cardiac tissues from male Wistar rats aged 3, 6, 12 and 24 months, and the expressions of p16, LC3?/?, ATG5 and ATG7 protein were detected in cardiac tissues in rats aged 3 months, 24 months and 24 months with spermidine administrtion, respectively. The myocardial autophagosome formation was observed by transmission electron microscope. In addition, cell cross-sectional area, cell apoptosis rate and generation of ROS were evaluated. Results With heart aging in rats, ODC expression and LC3?/? ratio were degraded, and SSAT and p62 protein expression upgraded. In rats aged 24 months, myocardium showed increased p16 expression, cell cross-sectional area, cell apoptosis and ROS generation, while cell autophagosome formation decreased, p62 expression increased, the expression of LC3?/?, ATG5 and ATG7 all declined. Spermidine intervention obviously inhibited myocardial changes induced by aging, showing decrease in cell cross-sectional area, apoptosis, ROS generation and p62 expression, and increase in LC3?/?, ATG5 and ATG7 expression. Conclusions With heart aging in rats, polyamine anabolism is degraded, catabolism is upgraded, and cell autophagy declined. Exogenous spermidine might delay aging through inducing autophagy of cardiomyocytes.
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Polyamines are a kind of aliphatic amines that exist widely in nearly all organisms. Polyamines interact with biological macromolecules through ionic interactions and hydrogen bonds, thereby they could affect the cell growth via regulating the function of macro-molecules. The impact of polyamines on nucleic acids has been thoroughly studied. However, their effects on protein structure and functions are not well established. This review summarizes the recent progress on how polyamines affect proteins, including metabolic enzymes, ion channel proteins and other important proteins. The interaction between polyamines and proteins is discussed, and the review also summarizes the challenges in studying polyamine-protein interaction as well as the potential application of these studies on the therapy of correlated diseases.
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@# Objective: To investigate the influence of inhibiting expression of polyamine-modulated factor (PMF-1) on the antitumor effect of glucocorticoid dexamethasone (DEX) in human cervical cancer Caski cells. Methods: siRNAs which target human PMF-1 gene were designed and synthesized, and their effect on the expression of PMF-1 in Caski cells was evaluated by Western blotting. The PMF-1 down-regulated and control Caski cells were treated with DEX, and then the affect of PMF-1 down regulation on the sensitivity of the tumor cells to DEX was analyzed. MTT method was used to detect cell proliferation, flow cytometry was used to analyze cell cycle, Western blotting method was used to evaluate expression level of glucocorticoids receptor (GR), and HPLC was used to analyze intracellular polyamine content. Results: The transient transfection of Caski cells with siRNAwhich targets PMF-1 gene can significantly reduce the expression level of PMF-1 protein. Compared with the control cells, treating PMF-1 down-regulated Caski cells with DEX can more effectively inhibit cell proliferation（P<0.01), up regulate GR expression, arrest cell cycle at G2 stage（P<0.01), and also significantly reduce intracellular polyamine level（P<0.01). Conclusion：Inhibiting PMF-1 expression can enhance antitumor pharmacological activity of DEX against human cervical cancer cells, and the underlying mechanism may be related with enhanced cell cycle inhibition and decreased intracellular polyamine level.
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OBJECTIVE: To study the effect of a novel naphthalimide-polyamine conjugate on apoptosis of human hepatocellular carcinoma HepG2 cell line. METHODS: MTT assay was used to evaluate the cell inhibition rate. The cellular morphous was detected with AO/EB/Hoechst staining, the ROS was detected with DCFH-DA staining, the mitochondrial membrane potential (ΔΨm) was detected with Rh123 staining, the expression of caspase-9 and PARP-1 was detected by immunofluorescence method. The cell cycle was detected by HCS. Apoptosis of HepG2 cells was quantified by flow cytometry using Annexin V/PI stain. RESULTS: Proliferation of HepG2 cells was inhibited significantly by the naphthalimide-polyamine conjugate in a dosage dependant manner. Under HCS, some HepG2 cells underwent a typical apoptotic morphologic change and the expression of ROS and caspase-9 was increased. The expression of PARP-1 and the mitochondrial membrane potential (ΔΨm) was detected decreased. Flow cytometry indicates 64.12% HepG2 cells were induced apoptosis after 48 h incubation with 25 μmol·L-1 naphthalimide-polyamine conjugate. CONCLUSION: The naphthalimide-polyamine conjugate could significantly induce the apoptosis of HepG2 cells by the mitochondrial pathway. The mechanism is concerned with increasing ROS, decreasing the mitochondrial membrane potential, upgrading caspase-9 and decreasing the expression of PARP-1.
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Objective: To study the effect of Astragalus membranaceus polysaccharides (AMP) on migration, intracellular polyamines content, cytosolic free Ca2+ concentration ([Ca2+]cyto), and RhoA protein expression of intestinal epithelial (IEC-6) cells, so as to explore the repairing mechanism of Astragalus membranaceus (AM) on gastrointestinal injury. Methods: AM was extracted by water and precipitated by ethanol, AM crude polysaccharides were obtained after removing protein. AMP I, II, III, and IV were obtained by DEAE cellulose column chromatography. AMP V was further obtained by Sephadex LH-20 gel column chromatography from AMP I. Cell migration model was established by Tips scratch method; High performance liquid chromatography was used to determine the polyamines content; Flow cytometry was used to detect the [Ca2+]cyto; Western blotting analysis was used to detect RhoA protein expression. The improving effect of AMP on migration, [Ca2+]cyto, and RhoA protein expression of normal and polyamine-depleted IEC-6 cells was observed. Results: AM crude polysaccharides, AMP I, and AMP V (100 mg /L or 200 mg /L) promoted cell migration and reversed the inhibition of cell migration induced by DFMO (P < 0.01); AMP V increased the intracellular polyamines content in normal and polyamine-deficient IEC-6 cells; AMP V also enhanced [Ca2+]cyto in the process of IEC-6 cell migration and reversed the reduction of [Ca2+]cyto induced by DFMO (P < 0.01); Further study suggested that AMP V increased RhoA protein expression in normal and polyamine-deficient IEC-6 cells (P < 0.01). Conclusion: These results indicate that the repairing effect of AM on the gastrointestinal mucosa damage may be related to its role of increasing polyamines content, then improving [Ca2+]cyto and RhoA protein expression and thereby promoting cell migration.
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Objective To investigate the effect of atorvastatin calcium on serum endothelin-1(ET-1), polyamine oxidase(PAO), heart-type fatty acid binding protein(H-FABP), vascular endothelial growth factor(VEGF), 100β, inflammatory cytokines and nerve function in patients with acute cerebral infarction.Methods According to the random number table, 113 patients were randomly divided into two groups (n=61) and control group (n=52).The control group received conventional treatment methods, and observation group received atorvastatin calcium on the basis of control group. The treatment course was two weeks.Serum ET-1, PAO, H-FABP, VEGF, S100β, inflammatory cytokines and NIHSS score were compared between two groups before treatment, 7d and 14d after treatment.Results The serum levels of ET-1, PAO, H-FABP after 7d, 14d treatment of observation group was significantly lower than that of control group, respectively (P<0.05).The VEGF level of observation group after 7 d, 14 d treatment of observation group was significantly higher than that of control group, respectively (P<0.05).The S100βlevel after 7 d, 14 d treatment of observation group was significantly lower than that of control group, respectively (P<0.05).The hs-CRP, IL-8, TNF-αlevels after 7 d, 14 d treatment of observation group was significantly lower than that of control group, respectively (P<0.05).NIHSS score after treatment of observation group was significantly lower than that of control group (P<0.05).Conclusion The atorvastatin calcium can improve neurological function in patients with brain injury through reducing serum ET-1, PAO, H-FABP and S100βlevels, promote angiogenesis through increasing VEGF expression, and alleviate inflammation and ischemia-reperfusion injury through reducing inflammatory cytokines, thereby promote neurological functional recovery.
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A series of genistein-polyamine conjugates (4a-4h) were designed, synthesized and evaluated as multi-functional anti-Alzheimer agents. The results showed that these compounds had significant cholinesterases (ChEs) inhibitory activity. Compound 4b exhibited the strongest inhibition to acetylcholinesterase (AChE) with an IC50 value of 2.75 μmol/L, which was better than that of rivastigmine (5.60 μmol/L). Lineweaver-Burk plot and molecular modeling study showed that compound 4b targeted both the catalytic active site (CAS) and the peripheral anionic site (PAS) of AChE. Besides, compound 4b showed potent metal-chelating ability. In addition, it was found that 4a-4h did not affect HepG-2 cell viability at the concentration of 10 μmol/L.
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Aim To study the effects of difluoromethy-lornithine (DFMO)on cardiomyocytes hypertrophy and apoptosis induced by isoproterenol (ISO).Methods The cardiomyocytes were divided into four groups ran-domly:Control group,ISO group,ISO+DFMO group and (ISO +DFMO +Putrescine)group.The hyper-trophic model of cardiomyocytes was induced by ISO, the cardiomyocytes were pretreated with 0.5 mmol · L-1 DFMO and 0.5 mmol·L-1 putrine,the gene ex-pression of ANP,the surface area of cardiomyocytes and the contents of LDH and MDA were measured. Apoptotic value of cardiomyocytes was observed by An-nexin V/PI,the gene and protein expressions of Bcl-2 and Bax were detected by real-time PCR and Western blot.Results Compared with ISO group,the cell sur-face area,the gene level of ANP,the apoptosis value of cardiomyocytes and the contents of LDH and MDA were decreased in ISO +DFMO group (P <0.05 ). Meanwhile,DFMO pretreatment upregulated the gene and protein expressions of Bcl-2 (P <0.05 ), in-creased the ratio of Bcl-2/Bax (P<0.05 ),downregu-lated the gene and protein levels of Bax (P<0.05 ). Conclusion The cardiomyocyte hypertrophy and ap-optosis induced by ISO can be protected by DFMO pre-treatment,which may be associated with the expression of apoptotic gene Bcl-2 and Bax.
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A giardíase é uma doença causada pelo protozoário flagelado Giardia lamblia, e sua sintomatologia é caracterizada pela eliminação de fezes esteatorréicas, dores abdominais e náuseas. Segundo o CDC estima-se que há cerca 1,2 milhões de casos por ano de giardíase, acometendo principalmente crianças em idade escolar. Atualmente, o tratamento da giardíase é realizado principalmente pelo uso do fármaco da família dos 5-nitromidazóis, metronidazol (Flagyl®), secnidazol e tinidazol em particular. Estes são confrontados em casos de resistência clínica causada pelo frequente uso inadequado do medicamento e/ou abandono do tratamento. Além disso, o metronidazol pode apresentar efeito carcinogênico em longo prazo em humanos. Desta forma, novos estudos com análogos e/ou inibidores de poliaminas podem levar à elucidação dos mecanismos de ação envolvidos, favorecendo o estabelecimento de novos regimes terapêuticos mais seguros e eficazes. Em nosso trabalho, foram testadas as substâncias ciclohexilamina (CHA) e o metronidazol que são produtos sintéticos, com o objetivo de avaliar os seus efeitos na proliferação celular, caracterização dos moduladores do metabolismo de poliaminas, avaliação nas mudanças no potencial redox e elucidação de seus possíveis mecanismos de ação nos trofozoítos de Giardia lamblia. Foi realizada uma avaliação da proliferação celular na presença de CHA para trofozoítos de Giardia lamblia, onde observamos que a substância demonstrou ter ação siginficativa apresentando um efeito dosedependente. Observamos que os trofozoítos de G. lamblia apresentam uma inibição significativa do crescimento em presença de concentrações milimolares do CHA, cujo IC50 em 72 horas foi de 1,646 mM...
Giardiasis is a disease caused by the flagellate protozoan Giardia lamblia, and its symptomatology is characterized by steatorrhea, abdominal pain and nausea. According to the CDC, an estimate number of 1.2 million cases of giardiasis happen every year, affecting especially schoolchildren.Nowadays, giardiasis treatment is based on drugs from the 5-nitroimidazole family, particularly metronidazole (Flagyl), secnidazole and tinidazole. Those drugs are indiscriminately used by the population, and it's not uncommon to find them causing clinical resistance due to inappropriate utilization and/or tratment abandon. Besides that, metronidazole can present longterm carcinogenic effect in humans. Thus, new studies with analogs and/or polyamines inhibitors can lead to the clarification of the drugs action mechanis, favouring the establishment of new, safer and more efficient therapeutic regimens.Our work tested cyclohexylamine (CHA) and metronidazole, wich are synthetic products, in order to evaluate their effects on cell proliferation and on changes in redox potential, characterize polyamines metabolism modulator and describe their possible action mechanisms on Giardia lamblia trophozoites. We evaluated Giardia lamblia trophozoites cell proliferation in the presence of CHA; it was observe that the substance shows significant action, presenting dose-dependent effect. We also observed that G. lamblia trophozoites presented significant growth inhibition when exposed to millimolar concentrations of CHA - its IC50 in 72 hours was 1,646mM...
Subject(s)
Humans , Giardia/immunology , Giardia/parasitology , Trophozoites/growth & development , Trophozoites/physiology , Trophozoites/immunology , Trophozoites/parasitology , Trophozoites/pathologyABSTRACT
Objective To investigate the effect of the new polyamine analogue tetrabutyl propanediamine (TBP) on cell proliferation and the underlying mechanism. Methods MTT assay was performed to determine cell proliferation. Flow cytometry was performed to detect cell cycle transition. DNA fragmentation and mitochondrial membrane potential determinations were performed to detect cell apoptosis. The activity of key enzymes in polyamine catabolism was detected by chemiluminescence assay. Results TBP could significantly inhibit the proliferation of HL-60 cells by blockingcell cycle transition and by inducing apoptosis. The TBP-induced apoptosis of HL-60 cells was in a dose-dependent manner. The enzyme activities of SMO and APAO were also significantly increased in HL-60 cells after treatment with 100 μM TBP for 24 hours. Conclusions TBP, as a new putrescine analogue, could inhibit proliferation of HL-60 cells by increasing the enzyme activity of SMO and APAO and inducing apoptosis.
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Rapid tumor cell growth depends on intracellular polyamine levels higher than those of normal cells. Intracellular polyamine depletion inhibits tumor cell proliferation and induces tumor cell apoptosis. Therefore, polyamine metabolism has recently been identified as an important target for anti-tumor therapy. This article briefly summarizes recent polyamine metabolism targeting, polyamine depletion within the tumor cells through a variety of methods, and the antitumor effects of the treatment.
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Objective: To clarify whether polyamine-mediated triterpenoid synthesis in birch (Betula platyphylla) suspension cells induced by fungal elicitor. Methods: Fungal elicitor (40 μg/mL), putrescine (Put, 1 mmol/L), and D-arginine (D-Arg, 2 mmol/L) were added to the eight-day-old suspension cell culture, the content changes of triterpenoid and free polyamines were analyzed by chemical colorimetry and HPLC. The effect of polyamine on triterpenoid synthesis in B. platyphylla suspension cells induced by fungal elicitor was analyzed by pharmacology and restoration experiment. Results: After the treatment of fungal elicitor or Put, the contents and yields of free polyamines and triterpenoid were increased. Among of them, the triterpenoid content was the highest after 24 h treatment, the increasing rates were 68.54% and 30.34%, respectively. The triterpenoid content was increased by the cotreatment of fungal elicitor and Put, but the increasing degree of yield was lower than that by the treatment of fungal elicitor alone. Compared with the fungal elicitor alone, the cotreatment of fungal elicitor and D-Arg decreased the triterpenoid content by 40.57% in the highest degree of decreasing after 24 h treatment. In restoration experiment, with the treatment time prolonging, the effect of fungal elicitor, Put, or cotreatment of fungal elicitor and D-Arg on triterpenoid synthesis was decreasing to the control level finally. Conclusion: Polyamine could mediate the triterpenoid synthesis in cells of B. platyphylla induced by fungal elicitor.
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Se destaca la actividad de las flavoenzimas como amino-oxidasas, que intervienen en el metabolismo de las aminas biogénicas como biorreguladores, especialmente en el crecimiento y la diferenciación celular. La clasificación de las amino-oxidasas incluye flavoenzimas y quinoenzimas. Se analizan las amino-oxidasas que son flavoproteínas, como las monoamino-oxidasas y las poliamino-oxidasas. Se discuten las isoformas, estructuras y función de ambas, sus sustratos e inhibidores, la expresión de MAO-A y MAO-B en tejidos humanos y sus implicancias clínicas. MAO plaquetaria es un biomarcador de desórdenes mentales y neurodegenerativos. Los inhibidores selectivos de MAO-A resultaron ser eficaces antidepresivos, mientras que algunos de MAO-B se utilizan en el tratamiento de enfermedades de Parkinson y de Alzheimer. La identificación de elevadas concentraciones de poliaminas en varias enfermedades, desde cáncer y psoriasis hasta infecciones parasitarias, hace que la manipulación de su metabolismo sea un blanco terapéutico o preventivo en ciertas enfermedades. Se discute además qué poliamino-oxidasas actúan en el metabolismo de las poliaminas en humanos, frente a las presentes en plantas, bacterias y protistas. Las poliaminas y las enzimas de su metabolismo desempeñan funciones relevantes en los procesos de envejecimiento y en algunas enfermedades, como cáncer, diabetes mellitus, accidentes cerebro-vasculares, insuficiencia renal y trastornos psiquiátricos.
The activity of flavoenzymes as amine oxidases involved in the metabolism of biogenic amines as bioregulators is highlighted, particularly for cell growth and differentiation. The classification of amine oxidases includes flavoenzymes and quinoenzymes. Amine oxidases that are flavoproteins, such as monoamine oxidases and polyamine oxidases, are analyzed herein. The isoforms, structures and functions of both enzyme families, their substrates and inhibitors, the expression of MAO-A and MAO-B in human tissues, and their clinical implications are discussed. Platelet MAO is a biomarker of mental and neurodegenerative disorders. Selective MAO-A inhibitors proved to be effective antidepressants, while some MAO-B inhibitors are used for treatment of Parkinson's and Alzheimer's diseases. The identification of high concentrations of polyamines in a variety of diseases, from psoriasis to cancer and parasitic infections, makes handling their metabolism a therapeutic or preventive target for the treatment of some diseases. Also polyamine oxidase activity on polyamine metabolism in humans, compared to those present in plants, bacteria and protists,is discussed. Polyamines and the enzymes involved in their metabolism play important roles in the aging processes, as well as in certain diseases such as cancer, diabetes mellitus, stroke, kidney failure, and defined psychiatric disorders.
Foi enfatizada a atividade de flavoenzimas como as amina oxidases envolvidas no metabolismo de aminas biogênicas como biorreguladores, especialmente no crescimento e diferenciação celular. A classificação das amina oxidases inclui flavoenzimas e quinoenzimas. Amina oxidases que são flavoproteínas, tais como monoamina oxidases e poliamina oxidases, são analisadas. Isoformas, estrutura e função das duas oxidases são discutidas, os seus substratos e inibidores, a expressão de MAO-A e MAO-B em tecidos humanos e suas implicações clínicas. MAO plaquetária é um biomarcador de desordens mentais e neurodegenerativas. Os inibidores selectivos da MAO-A resultaram ser eficazes antidepressivos, embora alguns dos MAO-B sejam utilizados no tratamento da doença de Parkinson e de Alzheimer. A identificação de elevadas concentrações de poliaminas em várias doenças, desde câncer e psoríase a infecções parasitárias, faz com que a manipulação do seu metabolismo seja um alvo terapêutico ou preventivo em certas doenças. Também se discute que a poliamina oxidase atua sobre o metabolismo das poliaminas no ser humano, em comparação com aquelas presentes em plantas, bactérias e protistas. As poliaminas e enzimas do seu metabolismo desempenham papéis relevantes nos processos de envelhecimento e em algumas doenças, tais como câncer, diabetes miellitus, acidente vascular cerebral, insuficiência renal e perturbações psiquiátricas.
Subject(s)
Monoamine Oxidase/biosynthesis , Monoamine Oxidase/metabolism , Monoamine Oxidase/physiology , Monoamine Oxidase Inhibitors , Polyamines , Polyamines/metabolismABSTRACT
Polyamines, putrescine, spermidine and spermine, are ubiquitous in living cells and are essential for eukaryotic cell growth. These polycations interact with negatively charged molecules such as DNA, RNA, acidic proteins and phospholipids and modulate various cellular functions including macromolecular synthesis. Dysregulation of the polyamine pathway leads to pathological conditions including cancer, inflammation, stroke, renal failure and diabetes. Increase in polyamines and polyamine synthesis enzymes is often associated with tumor growth, and urinary and plasma contents of polyamines and their metabolites have been investigated as diagnostic markers for cancers. Of these, diacetylated derivatives of spermidine and spermine are elevated in the urine of cancer patients and present potential markers for early detection. Enhanced catabolism of cellular polyamines by polyamine oxidases (PAO), spermine oxidase (SMO) or acetylpolyamine oxidase (AcPAO), increases cellular oxidative stress and generates hydrogen peroxide and a reactive toxic metabolite, acrolein, which covalently incorporates into lysine residues of cellular proteins. Levels of protein-conjuagated acrolein (PC-Acro) and polyamine oxidizing enzymes were increased in the locus of brain infarction and in plasma in a mouse model of stroke and also in the plasma of stroke patients. When the combined measurements of PC-Acro, interleukin 6 (IL-6), and C-reactive protein (CRP) were evaluated, even silent brain infarction (SBI) was detected with high sensitivity and specificity. Considering that there are no reliable biochemical markers for early stage of stroke, PC-Acro and PAOs present promising markers. Thus the polyamine metabolites in plasma or urine provide useful tools in early diagnosis of cancer and stroke.
Subject(s)
Animals , Humans , Mice , Acrolein , Biomarkers , Brain Infarction , C-Reactive Protein , Diacetyl , DNA , Early Detection of Cancer , Eukaryotic Cells , Hydrogen Peroxide , Inflammation , Interleukin-6 , Lysine , Metabolism , Oxidative Stress , Oxidoreductases , Phospholipids , Plasma , Polyamines , Putrescine , Renal Insufficiency , RNA , Sensitivity and Specificity , Spermidine , Spermine , StrokeABSTRACT
Objective: To analyze the effect of exogenous polyamine on the growth of Betula platyphylla suspension cells and triterpenoid accumulation. Methods: At the end of the growth stage, 0.1 mmol/L and 1.0 mmol/L putrescine (Put), spermine (Spm), and spermidine (Spd) were added to the suspension cells of B. platyphylla, the triterpenoid content and gene expression of lupeol synthase (LUS) related to the synthesis of triterpenoid were analyzed by chemical colorimetry and RT-PCR, respectively. Results: Except for the treatment with 1.0 mmol/L Spm, the cell viability, dry weight, and triterpenoid production in B. platyphylla suspension cells induced by Put and Spd were increased, and the dry weight and triterpenoid production of B. platyphylla cells were increasing with the extension of polyamine treatment time. Among them, dry weight and triterpenoid production were the highest after 2 d Put induction, which were 38.89% and 116.35% higher than those in the control, respectively. Triterpenoid production in B. platyphylla cells induced by polyamine was further confirmed by RT-PCR of LUS gene. Conclusion: Put could effectively enhance the growth of B. platyphylla suspension cells and triterpenoid accumulation.
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Objective: To observe the effect of flavonoid from Glycyrrhizae Radix (FGR) on the migration of small intestinal epithelial cells (IEC-6) and intracellular polyamines content. Methods: The cell migration model was constructed by scratch method. Under the intervention of α-difluoromethylornithine (DFMO), the specific inhibitor of polyamine synthesis, or 4-aminopyridine (4-AP), the specific inhibitor of potassium channel, the effect of FGR on cell migration was observed. The intracellular polyamine content being cultured for 12 or 24 h in normal and DFMO intervention condition was detected by precolumn derivatization-HPLC. Results: FGR could reverse the inhibitory effects of DFMO and 4-AP on cell migration, increase the intracellular content of polyamines after 12 or 24 h treatment, and reserve the decrease of polyamine content caused by DFMO. Conclusion: FGR could affect the polyamine-dependent signaling pathway, which may be related to the mechanisms for FGR promoting cell migration.
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Objective: To analyze the interaction between putrescine (Put) and hydrogen peroxide (H2O2) on the regulation of triterpenoid production in Betula platyphylla suspension cell culture. Methods: Put (1 mmol/L), H2O2 (0.1 mmol/L), and fungal elicitors (40 μg/mL) were added into the eight-day-old B. platyphylla suspension cell culture, and the changes of triterpenoid content, polyamine content, and H2O2 fluorescence intensity were analyzed by chemical colorimetry, HPLC, and fluorescence microscopy, respectively. Results: Put (1 mmol/L) increased the H2O2 fluorescence intensity and triterpenoid content in B. platyphylla suspension cell culture. H2O2 (0.1 mmol/L) promoted the triterpenoid production, inhibited the production of spermidine (Spd) and spermine (Spm), and had no effectt on Put content. Under the treatment of 1 mmol/L Put and 0.1 mmol/L H2O2, H2O2 fluorescence intensity and polyamine (PAs) content were between its separated treatment, and triterpenoid content was significantly higher than its separated treatment. Comparing with the fungal elicitor, fungal elicitor and H2O2 scavenger catalase (CAT) induced an increase of 0.62% and 16.05% in Put and Spd content, respectively, while no effect on Spm content. Fungal elicitor and PAs synthesis inhibitor D-arginine (D-arg) decreased the H2O2 fluorescence intensity. Under the treatment of fungal elicitor, the fluorescence intensity of CAT and D-arg, H2O2 and the contents of PAs and triterpenoid were lower than those of fungal elicitor and CAT or D-arg, but triterpenoid content was still higher than that of the control. Conclusion: This can be inferred that the interaction between Put and H2O2 regulates the triterpenoid production in birch suspension cell culture.