RESUMO
INTRODUCTION: Premature ovarian failure (POF) is defined as amenorrhea lasting for more than 4 months before 40 years of age, which is accompanied by a serum follicle-stimulating hormone (FSH) concentration above 40 mlU/mL. POF can cause a series of symptoms associated with low estrogen levels, such as hot flushes, excessive sweating, and infertility. This study aimed to report three cases of POF that were treated successfully with Korean medicine. CASE REPRESENTATION: Three patients with POF were selected for inclusion in this study. The treatment regimen consisted of herbal medicines, electroacupuncture, moxibustion, and Hominis placental pharmacopuncture. The basic treatment period was 3 months, and follow-up was performed after menstrual recovery. Following treatment, all three patients resumed menstruation without any adverse events. One patient also conceived successfully. CONCLUSIONS: These case reports suggest that Korean medicine could be effective for treating POF. Further preclinical and clinical studies are needed to investigate the mechanism of action of herbal medicines and acupuncture in improving menstruation and FSH levels.
Assuntos
Medicamentos de Ervas Chinesas , Insuficiência Ovariana Primária , Gravidez , Humanos , Feminino , Insuficiência Ovariana Primária/diagnóstico , Insuficiência Ovariana Primária/tratamento farmacológico , Placenta , Hormônio Foliculoestimulante/uso terapêutico , Medicamentos de Ervas Chinesas/uso terapêutico , República da CoreiaRESUMO
Several studies have documented the broad-spectrum bioactivities of a lotus seed (Plumula nelumbinis [PN]) green embryo extract. However, the specific bioactive components and associated molecular mechanisms remain largely unknown. This study aimed to identify the ion channel-activating mechanisms of PN extracts. Using fluorometric imaging and patch-clamp recordings, PN extracts were screened for calcium channel activation in dorsal root ganglion (DRG) neurons. The TRPV1 channels in DRG neurons were strongly activated by the PN extract (mean amplitude of 131 ± 45 pA at 200 µg/mL) and its purified glycosyloxyflavone narcissoside (401 ± 271 pA at 100 µM). Serial treatment with a 200 µg/mL PN extract in TRPV1-overexpressing HEK293T cells induced robust desensitization to 10 ± 10% of the initial current amplitude. Thus, we propose that the PN extract and narcissoside function as TRPV1 agonists. This new finding may advance our knowledge regarding the traditional and scientific functions of PN in human health and disease.
Assuntos
Gânglios Espinais , Extratos Vegetais , Canais de Cátion TRPV , Cálcio/metabolismo , Gânglios Espinais/metabolismo , Células HEK293 , Humanos , Lotus/química , Extratos Vegetais/farmacologia , Sementes/química , Células Receptoras Sensoriais/metabolismo , Canais de Cátion TRPV/agonistas , Canais de Cátion TRPV/genéticaRESUMO
Radiation proctitis is the collateral damage that occurs to healthy cells during radiation treatment of pelvic malignancies. Conservative treatment of radiation proctitis can mitigate inflammatory symptoms, but, to date, no therapeutic options are available for direct recovery of the damaged colonic epithelium. The present study assessed the ability of colon organoid-based regeneration to treat radiation proctitis. Radiation proctitis was induced in mice by irradiating their recta, followed by enema-based transplantation of mouse colon organoids. The transplanted colon organoids were found to successfully engraft onto the damaged rectal mucosa of the irradiated mice, reconstituting epithelial structure and integrity. Lgr5+ stem cells were shown to be pivotal to colon organoid mediated regeneration. Endoscopic examination showed the efficacy of localized transplantation of colon organoids with fibrin glue to irradiated sites. These findings provide useful insights into the use of colon organoid-based regenerative therapy for the treatment of radiation proctitis.
Assuntos
Proctite , Lesões por Radiação , Animais , Colo , Mucosa Intestinal , Camundongos , Organoides , Proctite/terapia , Lesões por Radiação/terapiaRESUMO
Coronavirus disease 2019 (COVID-19) has been a pandemic disease of which the termination is not yet predictable. Currently, researches to develop vaccines and treatments is going on globally to cope with this disastrous disease. Main protease (3CLpro) from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is one of the good targets to find antiviral agents before vaccines are available. Some flavonoids are known to inhibit 3CLpro from SARS-CoV which causes SARS. Since their sequence identity is 96%, a similar approach was performed with a flavonoid library. Baicalin, herbacetin, and pectolinarin have been discovered to block the proteolytic activity of SARS-CoV-2 3CLpro. An in silico docking study showed that the binding modes of herbacetin and pectolinarin are similar to those obtained from the catalytic domain of SARS-CoV 3CLpro. However, their binding affinities are different due to the usage of whole SARS-CoV-2 3CLpro in this study. Baicalin showed an effective inhibitory activity against SARS-CoV-2 3CLpro and its docking mode is different from those of herbacetin and pectolinarin. This study suggests important scaffolds to design 3CLpro inhibitors to develop antiviral agents or health-foods and dietary supplements to cope with SARS-CoV-2.
Assuntos
Infecções por Coronavirus/tratamento farmacológico , Flavonoides/química , Pneumonia Viral/tratamento farmacológico , Proteínas Virais/antagonistas & inibidores , Proteínas Virais/química , Antivirais/química , Betacoronavirus , COVID-19 , Desenho de Fármacos , Transferência Ressonante de Energia de Fluorescência , Humanos , Simulação de Acoplamento Molecular , Pandemias , Poliproteínas , Inibidores de Proteases/química , Ligação Proteica , Conformação Proteica , SARS-CoV-2 , Espectrofotometria , Triptofano/química , Tratamento Farmacológico da COVID-19RESUMO
Kummerowia striata (K. striata) is used as a traditional medicine for inflammation-related therapy. To determine whether it has beneficial anti-melanogenic and anti-oxidant activities, we investigated the biological activities of the ethanol extract of Kummerowia striata (EKS) using a variety of in vitro and cell culture model systems. The anti-melanogenic activity was assessed in B16F10 melanoma cells in terms of melanin synthesis and in vitro tyrosinase inhibitory activity. The anti-oxidant assays were performed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS). EKS showed strong anti-oxidant activities in DPPH and ABTS assays. The mRNA transcription levels and protein expression levels of tyrosinase, tyrosinase-related protein 1, tyrosinase-related protein 2, and microphthalmia-associated transcription factor decreased in a dose-dependent manner with EKS treatment. Additionally, EKS did not affect cell viability at different concentrations used in this study, indicating that the mechanism of action of EKS-mediated inhibition of melanin synthesis does not involve cytotoxicity. Also, we confirmed that p-coumaric acid and quercetin are important compounds for anti-melanogenesis and antioxidant properties of EKS. Collectively, our findings demonstrate for the first time that EKS possesses anti-melanogenic and anti-oxidant activities. Further evaluation and development of EKS as a functional supplement or cosmetic may be useful for skin whitening and reducing wrinkles.
RESUMO
An organoid is a complex, multi-cell three-dimensional (3D) structure that contains tissue-specific cells. Epithelial stem cells, which are marked by leucine-rich repeat-containing G-protein coupled receptor 5 (Lgr5), have the potential for self-renewal and expansion as organoids. However, in the case of intestinal organoids from Lgr5-EGFP-IRES-CreERT2 transgenic mice, in vitro expansion of the Lgr5 expression is limited in a culture condition supplemented with essential proteins, such as epidermal growth factor (E), noggin (N), and R-spondin 1 (R). In this study, we hypothesized that self-renewal of Lgr5+ stem cells in a 3D culture system can be stimulated by defined compounds (CHIR99021, Valproic acid, Y-27632, and A83-01). Our results demonstrated that dissociated single cells from organoids were organized into a 3D structure in the four compounds containing the ENR culture medium in a 3D and two-dimensional (2D) culture system. Moreover, the Lgr5 expression level of organoids from the ENR- and compound-containing media increased. Furthermore, the conversion of cultured Lgr5+ stem cells from 2D to 3D was confirmed. Therefore, defined compounds promote the expansion of Lgr5+ stem cells in organoids.
Assuntos
Organoides/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Células-Tronco Adultas/citologia , Células-Tronco Adultas/efeitos dos fármacos , Células-Tronco Adultas/metabolismo , Amidas/farmacologia , Animais , Autorrenovação Celular/efeitos dos fármacos , Autorrenovação Celular/genética , Autorrenovação Celular/fisiologia , Meios de Cultura Livres de Soro , Flavonoides/farmacologia , Expressão Gênica/efeitos dos fármacos , Humanos , Mucosa Intestinal/citologia , Mucosa Intestinal/metabolismo , Camundongos , Camundongos Transgênicos , Organoides/citologia , Organoides/crescimento & desenvolvimento , Pirazóis/farmacologia , Piridinas/farmacologia , Pirimidinas/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores Acoplados a Proteínas G/genética , Tiossemicarbazonas/farmacologia , Ácido Valproico/farmacologiaRESUMO
A Gram-negative, non-motile, aerobic, and rod-shaped bacterial strain designated as BR5-28T was isolated from the soil of a ginseng field at Baekdu Mountain Korea, and its taxonomic position was investigated using a polyphasic approach. Strain BR5-28T grew at 10-42°C (optimum temperature, 30°C) and pH 5.5-8.5 (optimum pH, 7.0) on R2A agar medium without additional NaCl supplementation. Strain BR5- 28T exhibited ß-glucosidase activity, which was responsible for its ability to transform the ginsenosides Rb1 and Rd (the two dominant active components of ginseng) to compound-K. Based on 16S rRNA gene phylogeny, the novel strain showed a new branch within the genus Mucilaginibacter of the family Sphingobacteriaceae, and formed clusters with Mucilaginibacter frigoritolerans FT22T (95.8%) and Mucilaginibacter gotjawali SA3-7T (95.7%). The G+C content of the genomic DNA was 45.1%. The predominant respiratory quinone was MK-7 and the major fatty acids were summed feature 3 (comprising C16:1 ω6c and/or C16:1 ω7c), iso-C15:0 and anteiso-C15:0. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Strain BR5-28T was differentiated genotypically and phenotypically from the recognized species of the genus Mucilaginibacter. The isolate therefore represents a novel species, for which the name Mucilaginibacter hankyongensis sp. nov. is proposed, with the type strain BR5-28T (=KCTC 22274T =DSM 21151T).
Assuntos
Bacteroidetes/classificação , Bacteroidetes/isolamento & purificação , Panax , Microbiologia do Solo , Técnicas de Tipagem Bacteriana , Bacteroidetes/enzimologia , Bacteroidetes/genética , Composição de Bases , DNA Bacteriano/análise , Ácidos Graxos/análise , Genótipo , Ginsenosídeos/metabolismo , Lipídeos/análise , Panax/crescimento & desenvolvimento , Panax/metabolismo , Fenótipo , Filogenia , RNA Ribossômico 16S , República da Coreia , Análise de Sequência de DNA , beta-Glucosidase/biossínteseRESUMO
Atractylenolide III (ATL-III) is an active compound of Atractylodes lancea, which has been widely used for the treatment of cancer. Cancer is closely connected with inflammation, and many anti-inflammatory agents are also used to treat cancer. We investigated the influence of ATL-III on thymic stromal lymphopoietin (TSLP)-induced inflammatory reactions. Pretreatment with ATL-III suppressed murine double minute 2 levels and promoted p53 levels in TSLP-treated human mast cell, HMC-1 cells. Mast cell proliferation increased by TSLP or IL-3 stimulation was significantly decreased by ATL-III pretreatment. Interleukin (IL)-13 and phosphorylated signal transducer and activator of transcription 3, 5, and 6 levels in TSLP-treated HMC-1 cells were also decreased by ATL-III pretreatment. In addition, ATL-III decreased the TSLP-induced production of proinflammatory cytokines (IL-6, IL-1ß, tumor necrosis factor-α, and IL-8). ATL-III decreased the levels of Bcl2 and procaspase-3 and increased caspase-3 activation and cleaved PARP levels. Furthermore, ATL-III decreased TSLP-induced mast cell proliferation and the production of inflammatory cytokine by LAD2 cells. Taken together, these findings suggest that ATL-III plays a useful role as an anti-inflammatory agent and should be viewed as a potential anti-cancer agent.
Assuntos
Atractylodes/química , Proliferação de Células/efeitos dos fármacos , Citocinas/farmacologia , Lactonas/farmacologia , Mastócitos/citologia , Extratos Vegetais/farmacologia , Sesquiterpenos/farmacologia , Anti-Inflamatórios/farmacologia , Caspase 3/genética , Caspase 3/metabolismo , Linhagem Celular , Humanos , Interleucina-13/genética , Interleucina-13/metabolismo , Mastócitos/efeitos dos fármacos , Mastócitos/metabolismo , Fosforilação , Proteínas Proto-Oncogênicas c-mdm2/genética , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Fator de Transcrição STAT6/genética , Fator de Transcrição STAT6/metabolismo , Linfopoietina do Estroma do TimoRESUMO
The ultrasonic treatment of varicose veins uses high-intensity focused ultrasound, in which a blood vessel is contracted by converting acoustic energy into thermal energy. In this study, we propose a phantom of varicose veins that can be applied for the efficient evaluation of ultrasonic treatment in varicose veins. The proposed phantom consisted of glycerol base tissue equivalent material, vessel mimic tube, and blood mimic substances. The vessel mimic tube was placed inner glycerol phantom and it was filled with blood mimic substances. Blood-mimicked substances are prepared by adjusting the concentration of the glycerol solution to be similar to the acoustic properties of the blood, and vessel-mimicking materials are selected by measuring acoustic properties and thermal shrinkage of various materials in a heat-shrinkable tube. The blood vessels surrounding the tissue are replaced with the phantom similar to glycerol-based organization, and venous blood flow is implemented using a DC motor. The heating characteristics according to the ultrasonic wave using the manufactured varicose veins phantom were evaluated. As the sound wave irradiation time and power increased, the contractility of the vessel mimicking materials and the temperature of the surrounding tissues were increased. When the blood-mimicking material was circulated, the highest temperature in the focused region and the contractility of vessel mimicking materials were reduced under the same conditions as used for sonication. The manufactured phantom may contribute to the treatment of varicose veins and can be used to predict the ultrasonic therapeutic efficiency of varicose veins.
Assuntos
Imagens de Fantasmas , Terapia por Ultrassom , Varizes/terapia , Humanos , Hipertermia InduzidaRESUMO
Amomum tsao-ko (A. tsao-ko) has been used as a traditional medicine for the treatment of infectious and digestive disorders. In the present study, we report the anti-inflammatory activity and molecular mechanism of 2,8-decadiene-1,10-diol (DDO) isolated from the extract of A. tsao-ko in lipopolysaccharide-stimulated RAW 264.7 cells. DDO treatment inhibited the production of nitric oxide and prostaglandin E2 by downregulating inducible nitric oxide synthase and cyclooxygenase-2 expression, respectively. Moreover, DDO suppressed the production of pro-inflammatory cytokines such as interleukin-6 and tumor necrosis factor-α. These inhibitory effects of DDO on the expression of inflammatory proteins were found to be mediated through the inactivation of mitogen-activated protein kinases (MAPKs) such as extracellular signal-regulated kinase, c-Jun-N-terminal kinase and p38(MAPK), and inhibition of nuclear factor-κB (NF-κB) pathways including degradation of inhibitor of κB-α and nuclear localization of NF-κB. Taken together, these findings demonstrate the pharmacological roles and molecular mechanisms of DDO in regulating inflammatory responses, and suggest further evaluation and development of DDO as a potent therapeutic agent for the treatment of inflammatory disorders.
Assuntos
Anti-Inflamatórios/farmacologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Álcoois Graxos/farmacologia , Inflamação/tratamento farmacológico , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Inibidor de NF-kappaB alfa/metabolismo , NF-kappa B/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Amomum/metabolismo , Animais , Linhagem Celular Transformada , Ciclo-Oxigenase 2/biossíntese , Dinoprostona/biossíntese , Regulação para Baixo/efeitos dos fármacos , Inflamação/patologia , Interleucina-6/biossíntese , Lipopolissacarídeos , Medicina Tradicional Coreana , Camundongos , NF-kappa B/metabolismo , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo II/biossíntese , Extratos Vegetais/farmacologia , Células RAW 264.7 , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Here we describe the generation of potato plants that constitutively overexpressed, UDP-N-acetylglucosamine:dolichol phosphate-N-acetylglucosamine-phosphotransferase (GPT). Such transgenic plants can be formed in a medium with tunicamycin at 9.8 ± 0.28% efficiency, similar to the 9.4 ± 1.10 for the bialaphos resistance gene (Bar) gene. This study indicated that GPT transformation was very stable with high reproducibility, and that growth and tuber production in the GPT-transformed plants were stronger than in the wild-type plants.
Assuntos
Engenharia Genética/métodos , Solanum tuberosum/genética , Transferases (Outros Grupos de Fosfato Substituídos)/genética , Transformação Genética , Marcadores Genéticos/genética , Vetores Genéticos/genética , Plantas Geneticamente ModificadasRESUMO
Dioscin is a kind of steroidal saponin isolated from the root bark of wild yam Dioscorea nipponica. We investigated the antifungal effect of dioscin against different fungal strains and its antifungal mechanism(s) in Candida albicans cells. Using the propidium iodide assay and calcein-leakage measurement, we confirmed that dioscin caused fungal membrane damage. Furthermore, we evaluated the ability of dioscin to disrupt the plasma membrane potential, using 3,3'-dipropylthiadicarbocyanine iodide [DiSC(3)(5)] and bis-(1,3-dibarbituric acid)-trimethine oxanol [DiBAC(4)(3)]. Cells stained with the dyes had a significant increase in fluorescent intensity after exposure to dioscin, indicating that dioscin has an effect on the membrane potential. To visualize the effect of dioscin on the cell membrane, we synthesized rhodamine-labeled giant unilamellar vesicles (GUVs) mimicking the outer leaflet of the plasma membrane of C. albicans. As seen in the result, the membrane disruptive action of dioscin caused morphological change and rhodamine leakage of the GUVs. In three-dimensional contour-plot analysis using flow cytometry, we observed a decrease in cell size, which is in agreement with our result from the GUV assay. These results suggest that dioscin exerts a considerable antifungal activity by disrupting the structure in membrane after invading into the fungal membrane, resulting in fungal cell death.
Assuntos
Antifúngicos/farmacologia , Candida albicans/efeitos dos fármacos , Dioscorea/metabolismo , Diosgenina/análogos & derivados , Biofísica/métodos , Membrana Celular/efeitos dos fármacos , Diosgenina/química , Relação Dose-Resposta a Droga , Citometria de Fluxo/métodos , Fluoresceínas/química , Potenciais da Membrana/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Microscopia de Fluorescência/métodos , Extratos Vegetais/metabolismo , Propídio/farmacologia , Rodaminas/farmacologia , Fatores de TempoRESUMO
This study established a new system for potato transformation using toxoflavin as selection agent and toxoflavin lyase (tflA) as selectable marker gene. Potato plants expressing tflA was successfully transformed on toxoflavin medium with 27% efficiency, similar to that for the hygromycin/hpt selection system. The transgenic potato expressing tflA also showed resistance to Burkholderia glumea infection.
Assuntos
Engenharia Genética/métodos , Marcadores Genéticos/genética , Liases/genética , Pirimidinonas/farmacologia , Solanum tuberosum/efeitos dos fármacos , Solanum tuberosum/genética , Transformação Genética/efeitos dos fármacos , Triazinas/farmacologia , Liases/metabolismo , Pirimidinonas/metabolismo , Triazinas/metabolismoRESUMO
Nitric oxide (NO) is a pleiotropic regulator, critical to numerous biological processes, including vasodilatation and macrophage-mediated immunity. Macrophages express inducible NO synthase (iNOS) and produce NO after lipopolysaccharide (LPS) stimulation. Gallotannins are water-soluble polyphenols with wide-ranging biological activities. Various chemical structures of gallotannins occurring in medicinal and food plants that are used worldwide showed several remarkable biological and pharmacological activities. In the present study, we examined the inhibitory effects of gallotannin 1,2,3,6-tetra-O-galloyl-beta-D-allopyranose (GT24) isolated from Euphorbia jolkini on the LPS-induced NO production and underlying mechanisms of action. GT24 dose-dependently decreased LPS-induced NO production and iNOS expression in J774A.1 macrophages. In addition, GT24 inhibited LPS-induced activation of nuclear factor (NF)-kappaB as indicated by inhibition of degradation of I-kappaBalpha, nuclear translocation of NF-kappaB, and NF-kappaB dependent gene reporter assay. Our results suggest that GT24 possesses an inhibitory effect on the LPS-induced inflammatory reaction.
Assuntos
Anti-Inflamatórios/farmacologia , Euphorbia/química , Taninos Hidrolisáveis/farmacologia , Macrófagos/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , Óxido Nítrico/biossíntese , Extratos Vegetais/farmacologia , Animais , Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/uso terapêutico , Transporte Biológico , Núcleo Celular , Relação Dose-Resposta a Droga , Genes Reporter , Taninos Hidrolisáveis/isolamento & purificação , Taninos Hidrolisáveis/uso terapêutico , Proteínas I-kappa B/metabolismo , Lipopolissacarídeos , Macrófagos/metabolismo , Camundongos , Inibidor de NF-kappaB alfa , Óxido Nítrico Sintase Tipo II/metabolismoRESUMO
BACKGROUND: Patatins encoded by a multi-gene family are one of the major storage glycoproteins in potato tubers. Potato tubers have recently emerged as bioreactors for the production of human therapeutic glycoproteins (vaccines). Increasing the yield of recombinant proteins, targeting the produced proteins to specific cellular compartments, and diminishing expensive protein purification steps are important research goals in plant biotechnology. In the present study, potato patatins were eliminated almost completely via RNA interference (RNAi) technology to develop potato tubers as a more efficient protein expression system. The gene silencing effect of patatins in the transgenic potato plants was examined at individual isoform levels. RESULTS: Based upon the sequence similarity within the multi-gene family of patatins, a highly conserved target sequence (635 nts) of patatin gene pat3-k1 [GenBank accession no. DQ114421] in potato plants (Solanum tuberosum L.) was amplified for the construction of a patatin-specific hairpin RNAi (hpRNAi) vector. The CaMV 35S promoter-driven patatin hpRNAi vector was transformed into the potato cultivar Desiree by Agrobacterium-mediated transformation. Ten transgenic potato lines bearing patatin hpRNA were generated. The effects of RNA interference were characterized at both the protein and mRNA levels using 1D and 2D SDS/PAGE and quantitative real-time RT-PCR analysis. Dependent upon the patatin hpRNAi line, patatins decreased by approximately 99% at both the protein and mRNA levels. However, the phenotype (e.g. the number and size of potato tuber, average tuber weight, growth pattern, etc.) of hpRNAi lines was not distinguishable from wild-type potato plants under both in vitro and ex vitro growth conditions. During glycoprotein purification, patatin-knockdown potato tubers allowed rapid purification of other potato glycoproteins with less contamination of patatins. CONCLUSION: Patatin-specific hpRNAi effectively suppressed the expression of a majority of patatin variants in potato tubers via the specific degradation of individual mRNAs of the patatin multi-gene family. More importantly, patatin-knockdown potato tubers appear to be an ideal host for the production of human therapeutic glycoproteins, because they eventually allow fast, easy purification of recombinant proteins, with less contamination from potato glycoprotein patatins.
Assuntos
Hidrolases de Éster Carboxílico/genética , Glicoproteínas/metabolismo , Glicoproteínas/uso terapêutico , Proteínas de Plantas/genética , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Engenharia de Proteínas/métodos , Interferência de RNA , Solanum tuberosum/fisiologia , Hidrolases de Éster Carboxílico/metabolismo , Melhoramento Genético/métodos , Glicoproteínas/genética , Humanos , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Plantas Geneticamente Modificadas/genéticaRESUMO
Transgenic potato plants (SS2 and SS4) that overexpressed a chloroplastic copper/zinc superoxide dismutase lily gene were utilized as an H(2)O(2)-inducible system in order to study the role of H(2)O(2) as a signaling molecule in the biosynthesis of ethylene. SS2 and SS4 plants grown in vitro under sealed microenvironment (SME) conditions displayed anomalous phenotypes including reduction of stem elongation, radial stem growth, and promotion of root hair formation in the generated root, which were similar to ethylene-induced responses. In addition, SS4 plants showed severe vitrification in developing leaves and elevated ethylene production under SME conditions. After the ethylene action inhibitor AgNO(3), 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase (ACO) inhibitor CoCl(2), and ACC synthase inhibitor L -aminoethoxyvinylglycine were added to the growth media, the anomalous phenotypes in SS4 plants reverted to their normal phenotype with a concurrent decrease in ethylene production. Northern blot analysis showed that ACO transcripts in SS4 plants were constantly at high levels under normal and SME conditions, indicating that a high level of H(2)O(2) in SS4 plants up-regulates ACO transcripts. Moreover, the direct treatment of H(2)O(2) in potato plants confirmed the elevated expression of the ACO gene. Taken together, these data suggest that the high concentration of H(2)O(2) in transgenic potato plants stimulates ethylene biosynthesis by activating ACO gene expression.
Assuntos
Etilenos/biossíntese , Peróxido de Hidrogênio/metabolismo , Lilium/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Solanum tuberosum/metabolismo , Superóxido Dismutase/metabolismo , Aminoácido Oxirredutases/metabolismo , Cobalto/farmacologia , Etilenos/antagonistas & inibidores , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Glicina/análogos & derivados , Glicina/farmacologia , Microscopia Eletrônica de Varredura , Compostos Organofosforados/farmacologia , Fenótipo , Caules de Planta/efeitos dos fármacos , Caules de Planta/genética , Caules de Planta/crescimento & desenvolvimento , Caules de Planta/metabolismo , Plantas Geneticamente Modificadas/efeitos dos fármacos , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Nitrato de Prata/farmacologia , Solanum tuberosum/efeitos dos fármacos , Solanum tuberosum/genética , Solanum tuberosum/crescimento & desenvolvimento , Superóxido Dismutase/genéticaRESUMO
A higher concentration of H2O2 was detected in the sense transgenic potato plant (SS4) with the lily chCu,ZnSOD sequence, whereas higher levels of O2(-) was detected in the antisense transgenic plant (SA1) than the WT plant. The elongation growth in SA1 was significantly inhibited by treatment with diphenyleneiodonium, an inhibitor of O2(-) generation, and promoted in the SS4 on treatment with herbicide methyl viologen, a generator of apoplastic O2(-) . Higher concentrations of GAs were detected during plant growth and the early stage of tuberization in SA1. Complete recovery of the above elongation growth and microtuberization pattern in transgenic plants following treatment of GA(3) or an inhibitor of gibberellin synthesis, paclobutrazol, indicate that these changes were mainly caused by active GA levels. In conclusion, a specific ROS (O2(-) ) acts as a signal transducer via GA biosynthetic pathways for the regulation of plant growth and tuber development of potato.
Assuntos
Proteínas de Plantas/metabolismo , Solanum tuberosum/crescimento & desenvolvimento , Solanum tuberosum/metabolismo , Superóxido Dismutase/metabolismo , Superóxidos/metabolismo , Giberelinas/genética , Giberelinas/metabolismo , Tubérculos/crescimento & desenvolvimento , Tubérculos/metabolismo , Espécies Reativas de Oxigênio/metabolismoRESUMO
Electroanalytical procedure for the determination of nephrotoxic aristolochic acid-I in the medicinal plant has been developed in the presence of potential interferences of lead and cadmium by square wave voltametry (SWV). Among the phosphate buffers of pH values at 5.0, 6.1, 6.5 and 7.0, the phosphate buffers of pH 6.1 yielded the most accurate analysis of AA-I in the presence of Pb2+ and Cd2+; Pb2+ was precipitated as Pb(HPO4) and did not appear in the SW voltammogram, while Cd2+ appeared at -0.564 V which was well resolved from AA-I at -0.416 V. When the Ip of AA-I was plotted vs. concentrations between 1.67 x 10(-8) M and 1.67 x 10(-6) M in the presence of Pb2+ and Cd2+, a linear calibration curve was obtained with a slope of 6 x 10(8) nA/M and a correlation coefficient of 0.9999. The present method was applied to determine AA in the dried natural products of Aristolochia contorta Bunge; Total AA in the dried root and the ripe fructus of Aristolochia contorta Bunge were found as 25 +/- 1 microg/g and 85 +/- 3 microg/g, respectively.
Assuntos
Aristolochia/química , Ácidos Aristolóquicos/análise , Calibragem , Eletroquímica , Eletrodos , Frutas/química , Estrutura Molecular , Extratos Vegetais/química , Raízes de Plantas/química , Padrões de Referência , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Madimadi, a Korean folk medicine, has been applied to treat rheumatoid arthritis (RA). However, its mechanisms of action have not been examined. The involvement of inflammatory cytokines, particularly TNF-alpha, IL-1beta, and IL-8, resulting in local inflammation in the pathogenesis of RA is now widely accepted. Madimadi dose-dependently inhibited TNF-alpha, IL-1beta, and IL-8 production from activated human mast cells (HMC-1). RT-PCR revealed inhibition of TNF-alpha and IL-1beta transcription in activated HMC-1. In addition, we confirmed potent inhibition of TNF-alpha and IL-1beta production by Madimadi using purified human blood PBMC from an active RA group, but not from healthy or disease control groups. These novel insights into the immunosuppressive action of Madimadi are likely to impact the clinical use of this agent.
Assuntos
Interleucina-1/biossíntese , Interleucina-8/biossíntese , Mastócitos/efeitos dos fármacos , Mastócitos/imunologia , Medicina Tradicional do Leste Asiático , Fator de Necrose Tumoral alfa/biossíntese , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/imunologia , Artrite Reumatoide/metabolismo , Estudos de Casos e Controles , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Medicina Herbária , Humanos , Interleucina-1/imunologia , Interleucina-8/imunologia , Coreia (Geográfico) , Mastócitos/citologia , Mastócitos/metabolismo , Fitoterapia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
BACKGROUND: Substances inducing apoptosis have shown efficacy in the treatment of cancers. Poncirus trifoliata (L.) Raf. (Rutaceae) fruits (PTF) has been used for the treatment of various cancers among Korean Oriental Medical doctors. METHODS: PTF-induced cytotoxicity of human leukemia HL-60 cells was monitored by the MTT assay. The apoptosis was determined by (a) apoptotic morphology in microscopy; (b) DNA fragmentation in electrophoresis and FACS analysis; and (c) activation of caspase-3 and poly-ADP-ribose polymerase (PARP) cleavage assay. RESULTS: The cytotoxic activity of PTF in HL-60 cells was increased in a concentration- and time-dependent manner. PTF caused the cell shrinkage, cell membrane blebbing, apoptotic body and DNA fragmentation. PTF-induced apoptosis is accompanied by the activation of caspase-3 and the specific proteolytic cleavage of PARP. However, PTF did not show cytotoxicity in normal peripheral blood mononuclear cells. CONCLUSIONS: Our novel finding provides evidence that PTF could be a candidate as an anti-leukemic agent through apoptosis of cancer cells.