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1.
Nutrients ; 10(3)2018 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-29543781

RESUMO

The aim of this study was to investigate the chondroprotective effect of a standardized extract (KBH-JP-040) of the Korean traditional herbs Kalopanax pictus Castor-Aralia, Hericium erinaceus (Bull.) Persoon, and Astragalus membranaceus Schischkin on in vivo and in vitro osteoarthritis (OA) models. Cultured rat chondrocytes were pre-treated with KBH-JP-040 (50, 100 and 200 µg/mL) for 1 h, then recombinant human IL-1α (rhIL-1α) for 24 h. For the in vivo model, rabbits (n = 60) were equally divided into experimental groups: normal control (NC), a collagenase-induced OA group, and OA groups treated with KBH-JP-040 (75, 100, and 150 mg/kg body weight) and celecoxib (Cx, 100 mg/kg) orally for 28 days. Treatment with KBH-JP-040 significantly attenuated inflammatory cytokines and matrix metalloproteinases (MMPs), suppressed the expression of IκBα, NF-κB, and JNK/p38 mitogen-activated protein (MAP) kinase, and upregulated aggrecan and collagen type-II expression in rhIL-1α-stimulated chondrocytes. Furthermore, the serum and synovial levels of inflammatory cytokines of rabbits also decreased in the treatment groups when compared with the OA group. Improved magnetic resonance imaging and histopathological findings further confirmed the therapeutic efficacy of KBH-JP-040 against OA. In conclusion, these results indicate that KBH-JP-040 possesses chondroprotective effects, suppressing inflammation and MMPs, and downregulating IκBα, NF-κB, and JNK/p38 MAP kinase-signaling pathways. This might be a potential therapeutic candidate for OA treatment.


Assuntos
Anti-Inflamatórios/farmacologia , Artrite Experimental/prevenção & controle , Astragalus propinquus , Basidiomycota , Condrócitos/efeitos dos fármacos , Articulações/efeitos dos fármacos , Kalopanax , Extratos Vegetais/farmacologia , Animais , Anti-Inflamatórios/isolamento & purificação , Artrite Experimental/induzido quimicamente , Artrite Experimental/diagnóstico por imagem , Artrite Experimental/metabolismo , Astragalus propinquus/química , Basidiomycota/química , Celecoxib/farmacologia , Células Cultivadas , Condrócitos/metabolismo , Colágeno/metabolismo , Colagenases , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Mediadores da Inflamação/metabolismo , Interleucina-1beta/farmacologia , Articulações/diagnóstico por imagem , Articulações/metabolismo , Kalopanax/química , Imageamento por Ressonância Magnética , Masculino , Metaloproteinases da Matriz/metabolismo , Fitoterapia , Extratos Vegetais/isolamento & purificação , Plantas Medicinais , Coelhos , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos
2.
Plant Cell Physiol ; 59(2): 319-330, 2018 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-29186583

RESUMO

Kalopanax septemlobus, commonly named the castor aralia tree, is a highly valued woody medicinal tree belonging to the family Araliaceae. Kalopanax septemlobus contains approximately 15 triterpenoid saponins primarily constituted of hederagenin aglycones. Hederagenin is a representative precursor for hemolytic saponin in plants. In the present study, transcriptome analysis was performed to discover genes involved in hederagenin saponin biosynthesis in K. septemlobus. De novo assembly generated 82,698 unique sequences, including 17,747 contigs and 64,951 singletons, following 454 pyrosequencing. Oxidosqualene cyclases (OSCs) are enzymes that catalyze the formation of diverse triterpene skeletons from 2,3-oxidosqualene. Heterologous expression of an OSC sequence in yeast revealed that KsBAS is a ß-amyrin synthase gene. Cytochrome P450 genes (CYPs) make up a supergene family in the plant genome and play a key role in the biosynthesis of sapogenin aglycones. In total, 95 contigs and 110 singletons annotated as CYPs were obtained by sequencing the K. septemlobus transcriptome. By heterologous expression in yeast, we found that CYP716A94 was ß-amyrin 28-oxidase involved in oleanolic acid production from ß-amyrin, and CYP72A397 was oleanolic acid 23-hydroxylase involved in hederagenin production from oleanolic acid. Engineered yeast co-expressing KsBAS, CYP716A94 and CYP72A397 produced hederagenin. Kalopanax septemlobus CYP72A397 is a novel CYP enzyme that synthesizes hederagenin aglycone from oleanolic acid as a single product. In conclusion, we characterized three genes participating in sequential steps for hederagenin biosynthesis from ß-amyrin, which are likely to play a major role in hederagenin saponin biosynthesis in K. septemlobus.


Assuntos
Sistema Enzimático do Citocromo P-450/genética , Perfilação da Expressão Gênica , Genes de Plantas , Kalopanax/enzimologia , Kalopanax/genética , Ácido Oleanólico/análogos & derivados , Proteínas de Plantas/genética , Saponinas/biossíntese , Biocatálise , Vias Biossintéticas/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Cromatografia Gasosa-Espectrometria de Massas , Regulação da Expressão Gênica de Plantas , Ácido Mevalônico/metabolismo , Ácido Oleanólico/biossíntese , Ácido Oleanólico/química , Filogenia , Proteínas de Plantas/metabolismo , Saccharomyces cerevisiae/metabolismo , Saponinas/química , Transcriptoma/genética
3.
Int J Nanomedicine ; 12: 4563-4578, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28790819

RESUMO

Recently, environment-friendly synthesis of gold nanoparticles (GNPs) has been extensively explored by biologists and chemists. However, significant research is still required to determine whether "eco-friendly" GNPs are beneficial to human health and to elucidate the molecular mechanisms of their effects on human cells. We used human neuronal SH-SY5Y cells to show that treatment with Kalopanacis Cortex extract-capped GNPs (KC-GNs), prepared via an eco-friendly, fast, one-pot synthetic route, protected neuronal cells against oxygen-glucose deprivation/reoxygenation (OGD/R)-induced damage. To prepare GNPs, Kalopanacis Cortex was used without any chemical reducing and stabilizing agents. Ultraviolet-visible spectroscopy showed maximum absorbance at 526 nm owing to KC-GN surface plasmon resonance. Hydrodynamic size (54.02±2.19 nm) and zeta potential (-20.3±0.04 mV) were determined by dynamic light scattering. The average diameter (41.07±3.05 nm) was determined by high-resolution transmission electron microscopy. Energy-dispersive X-ray diffraction spectroscopy and X-ray diffraction confirmed the presence of assembled GNPs. Fourier transform infrared analysis suggested that functional groups such as O-H, C-C, and C-N participated in KC-GN formation. Cell viability assays indicated that KC-GNs restored the viability of OGD/R-treated SH-SY5Y cells. Flow cytometry demonstrated that KC-GNs inhibited the OGD/R-induced reactive oxygen species production and mitochondrial membrane potential disruption. KC-GNs also inhibited the apoptosis of OGD/R-exposed cells. Western blot analysis indicated that the OGD/R-induced cellular apoptosis and simultaneous increases in the expression of cleaved caspase-3, p53, p21, and B-cell lymphoma 2-associated X protein were reversed by KC-GNs. The KC-GN-mediated protection against OGD/R-induced neurotoxicity was diminished by NRF2 and heme oxygenase-1 gene knockdowns. Collectively, these results suggested that KC-GNs exerted strong neuroprotective effects on human neuronal cells, which might be attributed to the attenuation of OGD/R-induced neuronal cell injury through the NRF2 signaling pathway.


Assuntos
Kalopanax/química , Fator 2 Relacionado a NF-E2/metabolismo , Nanopartículas/química , Fármacos Neuroprotetores/farmacologia , Extratos Vegetais/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Glucose/metabolismo , Ouro/química , Ouro/farmacologia , Química Verde , Heme Oxigenase-1/metabolismo , Humanos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Nanopartículas/administração & dosagem , Fármacos Neuroprotetores/administração & dosagem , Fármacos Neuroprotetores/química , Oxigênio/metabolismo , Extratos Vegetais/síntese química , Extratos Vegetais/química , Espécies Reativas de Oxigênio/metabolismo , Espectrometria por Raios X , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
4.
Planta ; 244(3): 753-60, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27376942

RESUMO

MAIN CONCLUSION: Deciduous ring-porous species in Japan shed all of their leaves under severe water stress before large vessels in earlywood are embolized, and embolization take place during winter. Water in deciduous ring-porous species is mainly conducted upward via large earlywood vessels of the current year. Water columns in large vessels are vulnerable to drought-induced and freeze stress-induced embolisms. Although a vulnerability curve can be created to estimate the hydraulic capacity of plants, it remains unclear why the loss of conductivity in potted plants of ring-porous species does not reach 100 % under severe drought stress. In this study, two deciduous ring-porous species in Japan (Kalopanax septemlobus and Toxicodendron trichocarpum) were used to explain the species-specific pattern in the water-conducting pathway of the stem. We monitored and visualized the spatial distribution of xylem embolisms in the stem of K. septemlobus saplings under drought stress and freeze stress using compact magnetic resonance imaging and cryo-scanning microscopy. In addition, we evaluated the water ascent in the stems of both species using a dye uptake method. Although embolisms of large vessels were observed under drought stress and in winter, all leaves were dropped to avoid fatal water loss after embolization of some large vessels. In contrast, all large vessels were embolized in winter. Larger-diameter vessels of latewood in T. trichocarpum tended to function in trees growing in the warm temperate zone. Thus, our results suggest that the unclear curve may be derived from a discrepancy between leaf water potential and actual water potential in the xylem under severe drought stress. The frequency of xylem embolisms in deciduous ring-porous species in Japan mainly depends on the number of freeze-thaw cycles.


Assuntos
Kalopanax/fisiologia , Toxicodendron/fisiologia , Água/fisiologia , Xilema/fisiologia , Secas , Congelamento
5.
Int J Oncol ; 48(1): 261-70, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26549816

RESUMO

The Kalopanax septemlobus leaf (Thunb.) Koidz. has been used as a traditional medicine herb for the treatment of various human diseases for hundreds of years. In this study, we investigated the mechanism underlying the inhibitory effects of an ethanol extract of K. septemlobus leaf (EEKS) on proliferation of HepG2 hepatocellular carcinoma cells. For this study, cell viability and apoptosis were evaluated using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay, DAPI (4,6-diamidino-2-phenylindole) staining, agarose gel electrophoresis, and flow cytometry. Measurements of the mitochondrial membrane potential (MMP), caspase activity assays and western blots were conducted to determine whether HepG2 cell death occurred by apoptosis. Treatment of HepG2 cells with EEKS concentration-dependently reduced cell survival while significantly increasing the ratio of apoptotic cells. EEKS treatment increased the levels of the death receptors (DRs), DR4 and DR5, and activated caspases, as well as promoting proteolytic degradation of poly(ADP-ribose)-polymerase associated with the downregulation of protein expression of members of the inhibitor of apoptosis protein family. Treatment with EEKS also caused truncation of Bid, translocation of pro-apoptotic Bax to the mitochondria, and loss of mitochondrial membrane permeabilization, thereby inducing the release of cytochrome c into the cytosol. However, treatment of HepG2 cells with a pan-caspase inhibitor reversed EEKS-induced apoptosis and growth suppression, indicating that EEKS appears to induce apoptosis though a caspase-dependent mechanism involving both intrinsic and extrinsic apoptotic pathways. In addition, the phosphorylation level of AMP-activated protein kinase (AMPK) was elevated when cells were exposed to EEKS. A specific inhibitor for AMPK attenuated the EEKS-induced activation of caspases, and consequently prevented the EEKS-induced apoptosis and reduction in cell viability. Overall, our findings suggest that EEKS inhibits the growth of HepG2 cells by inducing AMPK-mediated caspase-dependent apoptosis, suggesting the potential therapeutic application of EEKS in the treatment or prevention of cancers.


Assuntos
Proteínas Quinases Ativadas por AMP/biossíntese , Carcinoma Hepatocelular/tratamento farmacológico , Neoplasias Hepáticas/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Proteínas Quinases Ativadas por AMP/genética , Apoptose/efeitos dos fármacos , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/patologia , Inibidores de Caspase/administração & dosagem , Caspases/biossíntese , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Kalopanax/química , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/patologia , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Extratos Vegetais/química , Folhas de Planta/química , Proteína X Associada a bcl-2
6.
IET Nanobiotechnol ; 9(4): 220-5, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26224352

RESUMO

Manganese dioxide (MnO2) nanoparticles were synthesised by the reduction of potassium permanganate (KMnO4) using Kalopanax pictus leaf extract at room temperature. A transparent dark-brown colour appeared after the addition of K. pictus leaf extract to the solution of permanganate. The time course of the reduction of KMnO4and synthesis of MnO2 nanoparticles was monitored by means of UV-Vis spectra. The reduction of KMnO4occurred after addition of plant extract with disappearance of KMnO4specific peaks and emergence of peak specific for MnO2nanoparticles. MnO2nanoparticles showed absorption maxima at 404 nm. The electron dispersive X-ray spectroscopy analyses confirmed the presence of Mn and O in the sample. X-ray photoelectron spectroscopy revealed characteristic binding energies for MnO2nanoparticles. Transmission electron microscopy micrographs revealed presence of uniformly dispersed spherical shaped particles with average size of 19.2 nm. The selected area electron diffraction patterns revealed the crystalline nature of MnO2nanoparticles. Fourier transform-infrared spectroscopy spectra of pure MnO2show the occurrence of O-Mn-O vibrational mode at around 518 cm⁻¹. The phyto-synthesised MnO2nanoparticles showed degradation ability of dyes (congo red and safranin O) similar to chemically synthesised MnO2nanoparticles. This study shows simple and eco-friendly synthesis of MnO2nanoparticles by plant extract and their utilisation for dye degradation for the first time.


Assuntos
Kalopanax/química , Compostos de Manganês/química , Nanopartículas Metálicas/química , Óxidos/química , Extratos Vegetais/metabolismo , Biotecnologia , Compostos de Manganês/metabolismo , Nanotecnologia , Óxidos/metabolismo , Extratos Vegetais/química
7.
Appl Biochem Biotechnol ; 174(2): 587-601, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25085530

RESUMO

Silver nanoparticles (AgNPs) were synthesised using Kalopanax septemlobus plant leaf extracts. UV-visible spectrophotometric, Fourier-transform infrared, electron dispersive X-ray spectroscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analyses confirmed synthesis of AgNPs. TEM micrographs revealed presence of well-dispersed AgNPs predominantly of small size and different shapes with an average particle size of 30.8 nm. Antimicrobial susceptibility tests of AgNP treatments revealed variability in sensitivity of bacteria Bacillus cereus and Saccharophagus degradans under study. Minimum inhibitory concentration (MIC) values of the AgNPs for B. cereus and S. degradans were found to be 30 and 10 µg/mL, respectively. The mixed culture of B. cereus and S. degradans treated with AgNPs at 10 µg/mL showed increase in growth with time, suggesting survival of bacteria in liquid media. The plating of mixed culture before AgNP treatment showed presence of both bacteria, but 24-h-old mixed culture treated with AgNPs at the concentration of 10 µg/mL showed presence of B. cereus colonies. SEM micrographs revealed damage to S. degradans cells but no effect on B. cereus cells after AgNP treatment. Confocal microscopic observations of AgNP-treated mixed cultures by Nile blue A staining indicated intact polyhydroxyalkanoates producing flourescent cells of B. cereus but damage and deformities in S. degradans cells. This study suggests that AgNPs can selectively inhibit growth of S. degradans and retain B. cereus at MIC of S. degradans. This report is a case study for selective inhibition of one bacteria and growth of the other in a culture using plant-synthesized silver nanoparticles.


Assuntos
Alteromonadaceae/efeitos dos fármacos , Bacillus cereus/efeitos dos fármacos , Kalopanax/química , Nanopartículas Metálicas , Extratos Vegetais/química , Prata/química , Testes de Sensibilidade Microbiana , Microscopia Eletrônica , Espectrometria por Raios X , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier
8.
J Sep Sci ; 37(5): 505-14, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24376118

RESUMO

A method based on HPLC coupled with an evaporative light scattering detection and ESI quadrupole TOF MS was established for the quantification and identification of phenolics and triterpene saponins in Kalopanacis Cortex using a gradient elution of acetonitrile with 0.1% formic acid and water with 0.1% formic acid on an RP C18 column (4.6 × 250 mm, 5 µm). Diverse validation parameters, such as the linearity, LOD and LOQ, accuracy, precision, repeatability, and stability, were successfully obtained. Additionally, the efficiencies of different extraction methods were compared. The developed method was applied for the quantitative analysis of twelve representative metabolites in 61 Kalopanacis Cortex samples. The quantitation results showed that coniferin, kalopanaxsaponin C, septemlosides II, III, C, and D exhibited distinct regional patterns in Kalopanacis Cortex samples. These six compounds including one new triterpene saponin show potential as marker compounds for evaluating the quality of Kalopanacis Cortex and the geographical variation in its chemical composition.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/química , Kalopanax/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Cromatografia Líquida de Alta Pressão/instrumentação , Estrutura Molecular , Espectrometria de Massas por Ionização por Electrospray/instrumentação , Triterpenos/química
9.
Arch Pharm Res ; 36(3): 327-34, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23444041

RESUMO

Kalopanax pictus (Araliaceae) is a deciduous tree distributed in Korea, Japan, and China. The stem bark of K. pictus has been functionally used as a traditional crude drug for the treatment of various inflammatory diseases. In the present study, we describe the inhibitory effects of oleanane-type triterpenes and saponins isolated from the stem bark of K. pictus on production of pro-inflammatory cytokines in LPS-stimulated bone marrow-derived dendritic cells. Of the compounds tested, 16,23,29-trihydroxy-3-oxo-olean-12-en-28-oic acid (1), 4,23,29-trihydroxy-3,4-seco-olean-12-en-3-oate-28-oic acid (2), 3ß,6ß,23-trihydroxyolean-12-en-28-oic acid 28-O-ß-D-glucopyranoside (3), nipponogenin E (6), 3ß,6ß,23-trihydroxyolean-12-en-28-oic acid (7), and caulophyllogenin (19) significantly inhibited the production of IL-12 p40 and IL-6 with IC50 values ranging from 3.3 to 9.1 µM. Compounds 2, 3, 7, and 19 significantly suppressed the secretion of TNF-α with IC50 ranging from 8.8 to 20.0 µM. These data provide scientific support for the use of K. pictus stem bark and its triterpene and saponin components in the inhibition of pro-inflammatory cytokine secretion, including IL-12 p40, IL-6, and TNF-α, and for prevention and treatment of inflammatory diseases.


Assuntos
Citocinas/antagonistas & inibidores , Células Dendríticas/efeitos dos fármacos , Mediadores da Inflamação/antagonistas & inibidores , Kalopanax , Ácido Oleanólico/farmacologia , Saponinas/farmacologia , Animais , Medula Óssea/efeitos dos fármacos , Medula Óssea/metabolismo , Células Cultivadas , Citocinas/biossíntese , Células Dendríticas/metabolismo , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/toxicidade , Camundongos , Camundongos Endogâmicos C57BL , Ácido Oleanólico/química , Ácido Oleanólico/isolamento & purificação , Casca de Planta , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Saponinas/química , Saponinas/isolamento & purificação
10.
Cryo Letters ; 33(5): 402-10, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23224373

RESUMO

A cryopreservation protocol has been developed for embryogenic callus cultures of castor aralia (Kalopanax septemlobus), a deciduous tree which is widely used in oriental medicine and in landscape design. Three preculture treatments, four loading and six vitrification solutions were tested in a vitrification procedure. Preculture of embryogenic callus (EC) with high sucrose concentrations (up to 0.7 M) showed no effect on regrowth after cryopreservation. Loading for 20 min at ambient temperature improved regrowth of cryopreserved EC by 70-75 percent compared with non-loaded samples, regardless of the composition of the loading solution. Among vitrification solutions, the highest regrowth of 95-100 percent after cryopreservation was obtained after incubation of EC in a vitrification solution A3-80 percent comprising (w/v) 33.3 percent glycerol + 13.3 percent DMSO + 13.3 percent EG + 20.1 percent sucrose for 40 min at 0°C. Profiling of crystallization and recrystallization events using differential scanning calorimetry (DSC) confirmed that freezing injury was minimized in samples after loading and cryoprotection with this vitrification solution. Unlike many other papers, the droplet-vitrification protocol did not produce higher post-cryopreservation regrowth of Kalopanax EC, compared with the vitrification procedure. When samples are sufficiently cryoprotected during VS treatment, vitrification using cryovials may be preferred, since droplet-vitrification is more complex and requires skilled personnel. Cryopreserved callus grew rapidly and produced numerous somatic embryos, which developed similarly to embryos obtained from non-cryopreserved samples.


Assuntos
Criopreservação/métodos , Kalopanax/embriologia , Vitrificação , Varredura Diferencial de Calorimetria , Crioprotetores/química , Cristalização , Dimetil Sulfóxido/química , Glicerol/química , Kalopanax/crescimento & desenvolvimento , Sacarose/química
11.
Cryo Letters ; 33(4): 271-9, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22987238

RESUMO

Although an appropriate cryopreservation protocol is a prerequisite for basic studies and large-scale implementation as well as further cryopreservation studies, the process relies on trial and error. Among the vitrification-based cryopreservation techniques, droplet-vitrification produces high post-cryopreservation recovery. However, the protocol itself cannot solve the problems engaged in plant cryopreservation, prominently due to dehydration with cytotoxic vitrification solutions. This paper proposes a set of treatments to develop droplet-vitrification using a standard procedure associated with additional treatments and alternative vitrification solutions. The proposed standard protocol consists of a progressive preculture with 0.3 M sucrose for 31 h and with 0.7 M for 17 h, loading with vitrification solution C4-35% (17.5 percent glycerol + 17.5 percent sucrose, w/v) for 20 to 40 min, dehydration with vitrification solutions A3-90 percent (37.5 percent glycerol + 15% DMSO + 15 percent EG + 22.5 percent sucrose) for 10 to 30 min or B1-100 percent (PVS3) for 40 to 120 min at room temperature, cooling the samples using aluminum foil strips, rewarming by plunging into pre-heated (40 degree C) unloading solution (0.8 M sucrose) and further unloading for 20 to 60 min, depending on size and permeability of the materials. Using this systematic approach we can identify whether the material is tolerant or sensitive to chemical toxicity and to the osmotic stress of dehydration with vitrification solutions, thus revealing which is the main barrier in solution-based vitrification methods. Based on the sensitivity of samples we can design a droplet-vitrification procedure, i.e. preculture, loading, dehydration with vitrification solutions, cooling and rewarming. Using this approach, the development of appropriate droplet-vitrification protocol is facilitated.


Assuntos
Criopreservação/métodos , Crioprotetores/química , Células Vegetais/fisiologia , Fenômenos Fisiológicos Vegetais , Vitrificação , Chrysanthemum/citologia , Chrysanthemum/fisiologia , Crioprotetores/metabolismo , Alho/citologia , Alho/fisiologia , Kalopanax/citologia , Kalopanax/fisiologia , Osmose , Brotos de Planta/citologia , Brotos de Planta/fisiologia , Rubia/citologia , Rubia/fisiologia , Solanum tuberosum/citologia , Solanum tuberosum/fisiologia
12.
Chem Biol Interact ; 199(2): 106-11, 2012 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-22705713

RESUMO

Methyl 3,5-dicaffeoyl quinate (MDQ), an active compound present in Kalopanax pictus, Salicornia herbacea L., Aster oharai and Solidago virga-aurea var. gigantean, is a dicaffeoylquinic acid derivative esterified by methanol. Recent studies have revealed that MDQ possesses multiple pharmacological activities, such as antitumor, antioxidative and cytoprotective activities. To date, there has been no attempt to test the action of MDQ in melanocytes. In this study, we investigated the effect of MDQ on melanogenesis in B16F10 mouse melanoma cells. MDQ inhibited melanin production and tyrosinase activity in B16F10 mouse melanoma cells without a direct inhibitory effect on mushroom tyrosinase activity. Furthermore, we also found that MDQ decreased protein expression levels of microphthalmia-associated transcription factor (MITF) and tyrosinase in B16F10 melanin cells. Meanwhile, phosphorylation of p38 mitogen-activated protein kinase (p38 MAPK) was significantly reduced after 6h MDQ treatment, and this expression recovered at 48 h. The phosphorylation of extracellular signal-regulated kinase (ERK) was significantly enhanced at 12-48 h, whereas no effect was observed in the phosphorylation of Akt. In addition, MDQ treatment did not significantly alter the expression levels of total p38 MAPK, ERK, and Akt. Thus, it seems that inhibition of phospho-p38 MAPK and activation of phospho-ERK may lead to the suppression of melanogenesis in MDQ-treated B16F10 mouse melanoma cells.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Kalopanax/química , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Melanoma/tratamento farmacológico , Ácido Quínico/análogos & derivados , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Agaricales/enzimologia , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Melaninas/antagonistas & inibidores , Melaninas/metabolismo , Melanócitos/efeitos dos fármacos , Melanócitos/metabolismo , Melanoma/metabolismo , Camundongos , Fator de Transcrição Associado à Microftalmia/metabolismo , Monofenol Mono-Oxigenase/antagonistas & inibidores , Monofenol Mono-Oxigenase/genética , Fosforilação/efeitos dos fármacos , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia , Ácido Quínico/isolamento & purificação , Ácido Quínico/farmacologia
13.
Mol Ecol ; 21(15): 3823-38, 2012 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-22646502

RESUMO

We investigated the biogeographic history of Kalopanax septemlobus, one of the most widespread temperate tree species in East Asia, using a combined phylogeographic and palaeodistribution modelling approach. Range-wide genetic differentiation at nuclear microsatellites (G'(ST) = 0.709; 2205 samples genotyped at five loci) and chloroplast DNA (G(ST) = 0.697; 576 samples sequenced for 2055 bp at three fragments) was high. A major phylogeographic break in Central China corresponded with those of other temperate species and the spatial delineation of the two temperate forest subkingdoms of East Asia, consistent with the forests having been isolated within both East and West China for multiple glacial-interglacial cycles. Evidence for multiple glacial refugia was found in most of its current range in China, South Japan and the southernmost part of the Korean Peninsula. In contrast, lineage admixture and absence of private alleles and haplotypes in Hokkaido and the northern Korean Peninsula support a postglacial origin of northernmost populations. Although palaeodistribution modelling predicted suitable climate across a land-bridge extending from South Japan to East China during the Last Glacial Maximum, the genetic differentiation of regional populations indicated a limited role of the exposed sea floor as a dispersal corridor at that time. Overall, this study provides evidence that differential impacts of Quaternary climate oscillation associated with landscape heterogeneity have shaped the genetic structure of a wide-ranging temperate tree in East Asia.


Assuntos
Clima , Genética Populacional , Kalopanax/genética , Filogeografia , Evolução Biológica , Núcleo Celular/genética , DNA de Cloroplastos/genética , DNA de Plantas/genética , Extremo Oriente , Variação Genética , Geografia , Haplótipos , Kalopanax/classificação , Repetições de Microssatélites , Modelos Genéticos , Dados de Sequência Molecular , Filogenia , Árvores/classificação , Árvores/genética
14.
Phytother Res ; 26(4): 546-51, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21928370

RESUMO

The stem-bark of Kalopanax pictus (KP, family Araliaceae), which contains triterpenoid saponins, has been shown to exhibit anticarcinogenic, antiinflammatory, antirheumatoid and antidiabetic activities. In a preliminary study, a KP methanol extract demonstrated acetylcholinesterase activity in vitro and memory enhancement in scopolamine-treated mice. Therefore, we isolated acetylcholinesterase inhibitors, kalopanaxsaponins A and B, from a KP butanol (BuOH) fraction, measured acetylcholinesterase activity in vitro, and investigated their memory-enhancing effects in a passive avoidance test, Y-maze test and Morris water maze test. These constituents inhibited acetylcholinesterase activity and significantly reversed scopolamine-induced deficits. They also increased brain-derived neurotrophic factor (BDNF) and phosphorylated cAMP response element binding (p-CREB) protein expression but reduced TNF-α increased by scopolamine. Based on these findings, kalopanaxsaponins A and B may ameliorate memory deficits by inhibiting acetylcholinesterase activity and inducing BDNF and p-CREB expression.


Assuntos
Kalopanax/química , Transtornos da Memória/tratamento farmacológico , Ácido Oleanólico/análogos & derivados , Fitoterapia , Saponinas/uso terapêutico , Acetilcolinesterase/metabolismo , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Butanóis , Inibidores da Colinesterase/isolamento & purificação , Inibidores da Colinesterase/uso terapêutico , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Avaliação Pré-Clínica de Medicamentos , Ativação Enzimática , Aprendizagem em Labirinto/efeitos dos fármacos , Memória/efeitos dos fármacos , Transtornos da Memória/induzido quimicamente , Metanol , Camundongos , Camundongos Endogâmicos ICR , Nootrópicos/isolamento & purificação , Nootrópicos/uso terapêutico , Ácido Oleanólico/isolamento & purificação , Ácido Oleanólico/uso terapêutico , Fosforilação , Casca de Planta/química , Saponinas/isolamento & purificação , Escopolamina/efeitos adversos , Fator de Necrose Tumoral alfa/metabolismo
15.
Carbohydr Res ; 346(16): 2567-75, 2011 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-21996602

RESUMO

Kalopanax pictus (Araliaceae) is a deciduous tree that grows in East Asian countries. Its stem bark and leaves have been used in traditional medicine to treat rheumatic arthritis, neurotic pain, and diabetes mellitus. A phytochemical study on a methanol extract of the stem bark of K. pictus resulted in the isolation of three new compounds, 6ß,16α-dihydroxy-hederagenin 3-O-ß-D-glucuronopyranoside (1), 3-O-ß-D-glucuronopyranosyl-28-O-ß-D-glucopyranosyl-6ß,16α-dihydroxy-oleanolic acid (2), and 3-O-ß-D-galactopyranosyl(1→3)-α-L-arabinopyranosyl hederagenin 28-O-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl ester (3), along with eight known compounds (4-11). Their structures were established on the basis of chemical and spectroscopic methods (IR, 1D and 2D NMR, and HRESITOFMS). Compounds 1-6 and 8-10 upregulated PPARs transcriptional activity in a dose-dependent manner in HepG2 cells, with EC(50) values in the range 0.20-15.5 µM. Moreover, the specific PPAR transactivational effects of compounds 1-6 and 8-10 on separate PPAR subtypes, PPARα, -γ, and -ß(δ) were further investigated. Compounds 4, 5, 8, and 10 showed significant PPARα transactivational activity, with EC(50) values of 7.8, 8.0, 10.3, and 17.3 µM, respectively. Compounds 2, 4, 6, and 8-10 exhibited PPARγ dose-dependent transactivational activity, with EC(50) values of 14.7, 15.5, 14.8, 10.9, 17.1, and 16.3 µM, whereas compounds 8 and 10 significantly upregulated PPARß(δ) transcriptional activity, with EC(50) values of 15.7 and 17.7 µM, respectively.


Assuntos
Kalopanax/química , Receptores Ativados por Proliferador de Peroxissomo/genética , Casca de Planta/química , Extratos Vegetais/química , Saponinas/química , Ativação Transcricional/efeitos dos fármacos , Triterpenos/química , Configuração de Carboidratos , Sobrevivência Celular/efeitos dos fármacos , Genes Reporter , Células Hep G2 , Humanos , Luciferases/biossíntese , Luciferases/genética , Modelos Químicos , Extratos Vegetais/farmacologia , Regiões Promotoras Genéticas , Saponinas/farmacologia , Espectrometria de Massas por Ionização por Electrospray , Triterpenos/farmacologia
16.
Zhong Yao Cai ; 34(5): 716-8, 2011 May.
Artigo em Chinês | MEDLINE | ID: mdl-21954557

RESUMO

OBJECTIVE: To investigate the chemical constituents of Kalopanax septemlobus. METHODS: Chromatographic techniques including silica gel, gel, semi-preparative HPLC and PTLC as well as recrystallization were employed in the isolation and purification, and the structures were elucidated by spectral analysis and physical and chemical properties. RESULTS: 6 compounds were identified as liriodendrin (1), (-) -syringarenol (2), trans-coniferyl aldehyde (3), trans-caffeic acid (4), beta-daucosterol (5), beta-sitosterol (6). CONCLUSION: Compounds 2 -5 are obtained from this genus for the first time.


Assuntos
Aldeídos/isolamento & purificação , Furanos/isolamento & purificação , Kalopanax/química , Lignanas/isolamento & purificação , Raízes de Plantas/química , Acroleína/análogos & derivados , Aldeídos/química , Ácidos Cafeicos/química , Ácidos Cafeicos/isolamento & purificação , Furanos/química , Lignanas/química , Estrutura Molecular , Casca de Planta/química , Plantas Medicinais/química , Sitosteroides/química , Sitosteroides/isolamento & purificação
17.
J Nat Prod ; 74(9): 1908-15, 2011 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-21870831

RESUMO

Five new compounds, 16,23,29-trihydroxy-3-oxo-olean-12-en-28-oic acid (1), 4,23,29-trihydroxy-3,4-seco-olean-12-en-3-oate-28-oic acid (2), 3ß,6ß,23-trihydroxyolean-12-en-28-oic acid 28-O-ß-D-glucopyranoside (3), 3-O-[2,3-di-O-acetyl-α-L-arabinopyranosyl]hederagenin 28-O-α-L-rhamnopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranoside (4), and 3-O-[3,4-di-O-acetyl-α-L-arabinopyranosyl]hederagenin 28-O-α-L-rhamnopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranoside (5), as well as 10 known compounds (6-15), were isolated from the stem bark of Kalopanax pictus. Compounds 1-5 and 7-14 inhibited TNFα-induced NF-κB transcriptional activity in HepG2 cells in a dose-dependent manner, with IC50 values ranging from 0.6 to 16.4 µM. Furthermore, the transcriptional inhibitory function of these compounds was confirmed on the basis of decreases in COX-2 and iNOS gene expression in HepG2 cells. The structure-activity relationship of the compounds with respect to anti-inflammatory activity is also discussed.


Assuntos
Anti-Inflamatórios/isolamento & purificação , Anti-Inflamatórios/farmacologia , Kalopanax/química , Saponinas/isolamento & purificação , Saponinas/farmacologia , Triterpenos/isolamento & purificação , Triterpenos/farmacologia , Anti-Inflamatórios/química , Sequência de Bases , Inibidores de Ciclo-Oxigenase 2/química , Inibidores de Ciclo-Oxigenase 2/isolamento & purificação , Inibidores de Ciclo-Oxigenase 2/farmacologia , Células Hep G2 , Humanos , Coreia (Geográfico) , Estrutura Molecular , NF-kappa B/efeitos dos fármacos , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Casca de Planta/química , Saponinas/química , Relação Estrutura-Atividade , Triterpenos/química
18.
J Chromatogr A ; 1218(37): 6433-8, 2011 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-21835418

RESUMO

G-quadruplex DNA structure is considered to be a very attractive target for antitumor drug design due to its unique role in maintaining telomerase activities. Therefore, discovering ligands with high stability of G-quadruplex structure is of great interest. In this paper, high-performance liquid chromatography (HPLC) was used for fast screening of G-quadruplex ligands from the crude extract of Kalopanax septemlobus (Thunb.) Koidz, a traditional Chinese medicine. Four potent G-quadruplex ligands were firstly selected through HPLC by comparing the peak profiles and absorption intensity of the crude sample before and after interaction with G-quadruplex DNA. Then the target compounds were isolated and purified by high-speed countercurrent chromatography (HSCCC) for further confirmation of their stabilities of G-quadruplex by temperature-dependent circular dichroism (CD). Four compounds were isolated and identified as 2,4-dihydroxybenzoic acid (I), chlorogenic acid (II), caffeic acid (III) and 5-feruloylquinic acid (IV) each by MS and NMR. Finally, compound I, II, III were each proved to be potent G-quadruplex ligands by decreasing the peak intensity in HPLC chromatogram after complexation with G-quadruplex, which stabilize G-quadruplex by 7±2 °C, 10±2 °C, and 3±2 °C respectively, based on CD analyses. However, compound IV showed no G-quadruplex stability. The decrease of peak absorption intensity in HPLC chromatogram is the most important signal to find G-quadruplex ligands. This provides a very promising strategy for fast screening G-quadruplex ligands from natural plant extracts.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Ácidos Cumáricos/química , Medicamentos de Ervas Chinesas/química , Quadruplex G , Hidroxibenzoatos/química , Kalopanax/química , Antineoplásicos/química , Antineoplásicos/metabolismo , Ácidos Cumáricos/metabolismo , Distribuição Contracorrente , Medicamentos de Ervas Chinesas/metabolismo , Hidroxibenzoatos/metabolismo , Ressonância Magnética Nuclear Biomolecular
19.
Fish Shellfish Immunol ; 31(6): 801-7, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21782952

RESUMO

Feeding kelp grouper, Epinephelus bruneus (26.1 ± 1.4), with 0%, 0.1%, 1.0%, and 2.0% Kalopanax pictus extract-supplementation diets, for 30 days reduced mortality on being challenged intraperitoneally 100 µl with bacterium Vibrio alginolyticus (2.1 × 10(7) cfu ml(-1)) and ciliate parasite Philasterides dicentrarchi (2.3 × 10(7) ciliates ml(-1)). The red blood cells (RBC), white blood cells (WBC), haemoglobin, haematocrit, lymphocytes, and monocytes levels significantly increased in kelp grouper fed with all doses of K. pictus-supplementation diets and challenged with bacterium and parasite when compared to control. However, the levels of mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), neutrophils, and thrombocytes did not significantly. The phagocytic activity, complement activity, and antiprotease activity did not significantly change in kelp grouper fed with 0.1% K. pictus-supplementation diets and challenged with bacterium and parasite. The respiratory activity, lysozyme activity, bactericidal activity, total protein level, and myeloperoxidase levels significantly increased in kelp grouper fed with all the doses of K. pictus-supplementation diet and challenged with bacterium and parasite. However, α2-macroglobulin level significantly increased with 1.0% diet, but not with 0.1% and 2.0% diets. Therefore this study suggests that 1.0% and 2.0% K. pictus-supplementation diets positively protected and enhanced the immune system in kelp grouper E. bruneus against V. alginolyticus and P. dicentrarchi infection.


Assuntos
Aquicultura/métodos , Infecções por Cilióforos/veterinária , Doenças dos Peixes/tratamento farmacológico , Doenças dos Peixes/imunologia , Kalopanax/química , Perciformes , Extratos Vegetais/uso terapêutico , Vibrioses/veterinária , Análise de Variância , Animais , Contagem de Células Sanguíneas/veterinária , Infecções por Cilióforos/tratamento farmacológico , Infecções por Cilióforos/imunologia , Suplementos Nutricionais , Doenças dos Peixes/microbiologia , Doenças dos Peixes/parasitologia , Hematócrito/veterinária , Hemoglobinas/análise , Macrófagos/metabolismo , Oligoimenóforos , Vibrioses/tratamento farmacológico , Vibrioses/imunologia , Vibrio alginolyticus
20.
Heredity (Edinb) ; 107(4): 338-48, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21427749

RESUMO

We aimed to reveal the effects of range expansion and subsequent lineage admixture from separated glacial refugia on genetic diversity of Kalopanax septemlobus in Japan, by combining nuclear microsatellite data and ecological niche modelling. Allelic richness and gene diversity were compared at the population and regional level. We also statistically examined these indices as a function of population accessibility to the last glacial maximum (LGM) palaeodistribution reconstructed by ecological niche modelling to test a simple range expansion scenario from glacial refugia. Genetic diversity was highest in the populations of southern Japan and gradually decreased towards the north. However, an additional centre of genetic diversity, when measured as gene diversity, was found in northern Honshu Island, where distinct lineages were shown to be in contact. Positive effects of population accessibility to the LGM range were detected in both diversity indices at different spatial scales. The combined data support independent postglacial range expansions towards the north from the edge populations on the exposed coastal shelf of Pacific and Sea of Japan in northern Honshu during the LGM, which subsequently resulted in markedly low genetic diversity in the northernmost extant range, Hokkaido. The regional increase in gene diversity in northern Honshu is likely to be the result of postglacial lineage admixture. Relative difference in the spatial scales best relating population genetic diversity with the LGM distribution can be explained by a higher rate of allelic richness diversity loss during range expansions and stronger effects of lineage admixture on gene diversity.


Assuntos
Evolução Molecular , Ligação Genética , Variação Genética , Kalopanax/genética , Camada de Gelo , Japão , Kalopanax/classificação , Repetições de Microssatélites , Filogenia
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