RESUMO
A rapid, sensitive, and selective liquid chromatography-tandem mass spectrometry was developed for the simultaneous determination of lycorine and galanthamine, two major constituents in Lycoris radiata extract, in rat plasma. Liquid-liquid extraction with ethyl ether was carried out using diphenhydramine as the internal standard. The two bioactive alkaloids were separated on a Zorbax SB-C18 reserved-phase column (150 mm × 4.6 mm, i.d., 5 μm) by gradient elution using a mobile phase consisting of methanol with 0.1% formic acid (A) and water with 0.1% formic acid (B) at a flow rate of 0.6 mL/min. All analytes showed good linearity over a wide concentration range (r (2)>0.99) and the lower limit of quantification was 3.00 ng/mL for each analyte. The average extraction recovery of the analytes from rat plasma was more than 82.15%, and the intra-day and inter-day accuracy and precision of the assay were less than 12.6%. The validated method was successfully applied to monitoring the concentrations and pharmacokinetic studies of two Amaryllidaceous alkaloids in rat plasma after an oral administration of Lycoris radiata extract.
Assuntos
Animais , Masculino , Ratos , Alcaloides de Amaryllidaceae , Farmacocinética , Cromatografia Líquida , Galantamina , Farmacocinética , Lycoris , Química , Parassimpatomiméticos , Farmacocinética , Fenantridinas , Farmacocinética , Extratos Vegetais , Química , Farmacocinética , Farmacologia , Ratos Wistar , Espectrometria de Massas em Tandem , MétodosRESUMO
In order to investigate the effects of phenylalanine, tyrosine and tyramine on the growth of Lycoris radiata suspension cells and the accumulation of alkaloids, the growth quantity of the cells as well as the content of alkaloids in cells were determined, which were treated with above three kinds of precursors alone and phenylalanine combined with tyrosine respectively. The results indicate that the addition of phenylalanine alone and addition of phenylalanine on the basis of tyrosine at high concentration (200 micromol/L) had no significant effect on the growth of Lycoris radiata suspension cells and the content of alkaloids in cells; whereas tyrosine and tyramine promoted the growth of the cells and alkaloids accumulation. Treated with tyrosine at high concentration (200 micromol/L), the content of alkaloids of the cells was 2.56-fold higher than that of the control group, the amounts of lycoramine (3.77 mg/g) and galanthamine (4.46 mg/g) were 6.61-fold and 6.97-fold higher than that of the control group, respectively. When treated with tyramine (200 micromol/L), the amount of alkaloids in Lycoris radiata suspension cells was 2.63-fold higher than that of the control group, and the amounts of lycoramine (4.45 mg/g) and galanthamine (5.14 mg/g) were 9.08-fold and 9.18-fold higher than that of the control group, respectively. The above results demonstrate that adding tyrosine and tyramine in the media significantly promoted the growth of the Lycoris radiata suspension cells and alkaloids accumulation in the cells.
Assuntos
Alcaloides de Amaryllidaceae , Química , Células Cultivadas , Meios de Cultura , Química , Galantamina , Química , Lycoris , Química , Fenilalanina , Química , Células Vegetais , Química , Extratos VegetaisRESUMO
Ten Amaryllidaceae alkaloids were isolated from the bulbs of Lycoris radiata. Their structures were identified as oxovittatine (1), apohaemanthamine (2), 9-O-demethylhomolycorine N-oxide (3), incartine (4), ismine (5), 6-O-methylpretazettine (6), tazettine (7), ungeremine (8), homolycorine (9), and O-methyllycorenine (10) by spectroscopic data analyses. Compound 1 was a new natural product. Compounds 2 and 3 were reported form the genus Lycoris for the first time and compounds 4-6 were isolated form the title plant for the first time.
Assuntos
Alcaloides , Química , Lycoris , Química , Extratos Vegetais , Química , Raízes de Plantas , QuímicaRESUMO
AIM@#To study the Amaryllidaceae alkaloids of the bulbs of Lycoris radiata.@*METHODS@#The chemical constituents were isolated and purified by various chromatographic techniques, and the chemical structures were elucidated on the basis of spectroscopic methods. In addition, the antiviral activities of alkaloids 1-10 were evaluated using flu virus A.@*RESULTS@#One new homolycorine-type alkaloid 2α-methoxy-6-O-ethyloduline (1), together with nine known alkaloids 2α-methoxy-6-O-methyloduline (2), trispherine (3), 8-O-demethylhomolycorine (4), homolycorine (5), 9-O-demethylhomolycorine (6), oduline (7), lycorenine (8), 6α-O-methyllycorenine (9) and O-ethyllycorenine (10) were obtained.@*CONCLUSION@#Alkaloid 1 is a new compound, and 1-3 were major alkaloids in this plant. Alkaloids 1-3 showed weak antiviral activities against flu virus A with IC50 values of 2.06, 0.69, and 2.71 μg·mL-1 and CC50 values of 14.37, 4.79, and 80.12 μg·mL-1, respectively.
Assuntos
Alcaloides , Química , Farmacologia , Antivirais , Química , Farmacologia , Flores , Química , Vírus da Influenza A , Lycoris , Química , Estrutura Molecular , Extratos Vegetais , Química , FarmacologiaRESUMO
In this study, we investigated the effect of methanolic extract isolated from the root of Lycoris aurea (LA) on the growth of cancer cells and the tube formation activity of endothelial cells. Various cancer cells were treated with LA at doses of 0.3, 1, 3, 10 or 30 microg/ml and LA significantly suppressed the growth of several cancer cell lines, including ACHN, HCT-15, K-562, MCF-7, PC-3 and SK-OV-3, in a dose-dependent manner. We also found that LA induced cell cycle arrest at G2/M phase in ACHN renal cell adenocarcinoma cells. Further study demonstrated that LA concentration-dependently inhibited the tube formation, which is a widely used in vitro model of reorganization stage of angiogenesis, in human umbilical vein endothelial cells. Collectively, these results show that LA inhibits the growth of cancer cells and tube formation of endothelial cells and the growth-inhibitory effect of LA might be mediated, at least in part, by blocking cell cycle progression.
Assuntos
Carcinoma de Células Renais , Ciclo Celular , Pontos de Checagem do Ciclo Celular , Linhagem Celular , Células Endoteliais , Células Endoteliais da Veia Umbilical Humana , Lycoris , MetanolRESUMO
<p><b>OBJECTIVE</b>To identify some closely related Lycoris species and evaluate interspecific relationships among them.</p><p><b>METHOD</b>The cpDNA trnL-F sequence of 20 taxa representing 15 species of Lycoris and Narcissus tazaetta var. chinensis as one out-group were determined by using direct sequencing of PCR product, and they were analyzed by means of the software of CLUSTRAL and MEGA.</p><p><b>RESULT</b>The length of trnL-F of all taxa was 905 - 1 036 bp. When the gaps were always treated as missing, there were 14 variable sites and 10 parsim-info sites, which could be used to identify some species of Lycoris. Four nucleotides inserteions/deletions were significant different among Lycoris and two species of Narcissus. Phylogeny tree was constructed with the maximum parsimony methods by bootstrap test. Three infrageneric clades of all Lycoris species were resolved. The classification was basically consistent with that of morphology except for L. longituba, L. aurea, and L. straminea.</p><p><b>CONCLUSION</b>The tree suggested that L. anhuiensis can not be taken as an independent species, while it may be of a variety or a hybrid of L. longituba. Regarding the hybrid origin species, the materal parent of L. rosea and L. haywardii was revealed to be L. sprengeri. There were some variations in the trnL-F sequence, which were good molecular markers for identification species of Lycoris.</p>
Assuntos
Análise por Conglomerados , Medicamentos de Ervas Chinesas , Genes de Plantas , Genética , Mutação INDEL , Lycoris , Classificação , Genética , Mutação , Nucleotídeos , Genética , Filogenia , Análise de Sequência de DNAAssuntos
Alelos , Impressões Digitais de DNA , Primers do DNA , DNA de Plantas , Genes de Plantas , Variação Genética , Genética Populacional , Lycoris/genética , Técnicas de Amplificação de Ácido Nucleico , Folhas de Planta/química , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , Especificidade da Espécie , Taq Polimerase/metabolismo , Moldes GenéticosRESUMO
<p><b>OBJECTIVE</b>To establish a method for determination of galanthamine in Lycoris radiata.</p><p><b>METHOD</b>HPLC separation was carried on a column of ODS (4.6 mm x 150 mm, 5 microm), with the mobile phase of phosphate buffer (pH = 3-4)-methanol (93:7) at the flow rate of 1.0 mL x min(-1), and the detection wavelength was set at 289 nm.</p><p><b>RESULT</b>Galanthamine revealed linearity within the range of 3-30 microg x mL(-1) (r = 0.9997), the detection limit was 0.3 ng. The average recovery was 99.5% (RSD = 0.5%).</p><p><b>CONCLUSION</b>The method is easy to operate and the results of the determination are accurate, it can be used to evaluate the quality of L. radiata.</p>
Assuntos
Cromatografia Líquida de Alta Pressão , Métodos , Galantamina , Lycoris , Química , Raízes de Plantas , Química , Plantas Medicinais , Química , Controle de QualidadeRESUMO
To obtain a primary overview of gene diversity and expression pattern in Lycoris longituba, 4,992 ESTs (Expressed Sequence Tags) from L. longituba bud were sequenced and 4,687 cleaned ESTs were used for gene expression analysis. Clustered by the PHRAP program, 967 contigs and 1,343 singlets were obtained. Blast search showed that 179 contigs and 227 singlets (totally 1,066 ESTs) had homologues in GenBank and 3,621 ESTs were novel.