We have recently discovered that brassinosteroids (BRs) can inhibit the growth of etiolated pea seedlings dose-dependently in a similar manner to the 'triple response' induced by ethylene. We demonstrate here that the growth inhibition of etiolated pea shoots strongly correlates with increases in ethylene production, which also responds dose-dependently to applied BRs. We assessed the biological activities of two natural BRs on pea seedlings, which are excellent material as they grow rapidly, and respond both linearly and uni-phasically to applied BRs. We then compared the BRs' inhibitory effects on growth, and induction of ethylene and ACC (1-aminocyclopropane-1-carboxylic acid) production, to those of representatives of other phytohormone classes (cytokinins, auxins, and gibberellins). Auxin induced ca. 50-fold weaker responses in etiolated pea seedlings than brassinolide, and the other phytohormones induced much weaker (or opposite) responses. Following the optimization of conditions for determining ethylene production after BR treatment, we found a positive correlation between BR bioactivity and ethylene production. Finally, we optimized conditions for pea growth responses and developed a new, highly sensitive, and convenient bioassay for BR activity.
- MeSH
- aminokyseliny cyklické metabolismus MeSH
- biotest metody MeSH
- brassinosteroidy farmakologie MeSH
- ethyleny metabolismus MeSH
- hrách setý účinky léků růst a vývoj metabolismus MeSH
- inhibitory růstu farmakologie MeSH
- kyseliny indoloctové farmakologie MeSH
- regulátory růstu rostlin farmakokinetika farmakologie MeSH
- semenáček účinky léků růst a vývoj metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Fifteen Amaryllidaceae alkaloids (1-15) of various structural types were isolated by standard chromatographic methods from fresh bulbs of Narcissus poeticus cv. Pink Parasol. The chemical structures were elucidated by MS, and 1D and 2D NMR spectroscopic analyses, and by comparison with literature data. Narcipavline (5) and narcikachnine (6) are reported here for the first time. In their structure are combined two basic structural types of Amaryllidaceae alkaloids (galanthamine- and galanthindole-structural types), which represent a new structural type of these compounds. Alkaloids isolated in sufficient amounts were evaluated for their human erythrocytic acetylcholinesterase, and human serum butyrylcholinesterase (HuBuChE) inhibition activity using Ellman's method. Z-Gly-Pro-p-nitroanilide was used as substrate in the prolyl oligopeptidase (POP) assay. Untested alkaloids were also screened for their cytotoxic activity against a small panel of human cancer cells, which spanned cell lines from different tissue types. In parallel, MRC-5 human fibroblasts were employed to determine overall toxicity against noncancerous cells. Some compounds were evaluated for their antiprotozoal activity. The newly isolated alkaloid narcipavline (5) showed interesting HuBuChE inhibition activity (IC50 = 24.4 ± 1.2 µM), and norlycoramine (11) demonstrated promising POP inhibition (IC50 = 0.21 ± 0.01 mM).
- MeSH
- alkaloidy chemie izolace a purifikace farmakologie MeSH
- buňky A549 MeSH
- buňky HT-29 MeSH
- butyrylcholinesterasa metabolismus MeSH
- cholinesterasové inhibitory chemie izolace a purifikace farmakologie MeSH
- HeLa buňky MeSH
- inhibitory růstu chemie izolace a purifikace farmakologie MeSH
- Jurkat buňky MeSH
- kořeny rostlin MeSH
- lidé MeSH
- MFC-7 buňky MeSH
- myši MeSH
- Narcissus * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
OBJECTIVE: To determine whether inhibition of epidermal growth factor (EGF) receptor tyrosine kinase with lapatinib affects oocyte maturation, expression of the cumulus expansion-associated genes such as tumor necrosis factor alpha-induced protein 6 (TNFAIP6) and prostaglandin-endoperoxide synthase 2 (PTGS2), and synthesis of hyaluronan (HA) and progesterone (P) by porcine oocyte cumulus complexes (OCC). DESIGN: Our work focuses on lapatinib, an orally active small molecule that selectively inhibits the tyrosine kinase domain of both EGF receptor and human EGF receptor 2, and downstream signaling. SETTING: A reproductive biology laboratory. PATIENT(S): Not applicable. INTERVENTION(S): Porcine OCC were cultured in vitro in a medium with FSH/LH in the presence/absence of lapatinib. MAIN OUTCOME MEASURE(S): Methods performed: real-time reverse transcriptase-polymerase chain reaction (PCR), immunofluorescence, RIA. RESULT(S): In FSH/LH-stimulated and expanded cumulus oophorus extracellular matrix, HA was detected with biotinylated HA-binding proteins. However, weaker HA- and weaker cytoplasmic TNFAIP6 were detected were detected in lapatinib-pretreated OCC. The expression of the two cumulus expansion-associated gene transcripts was significantly decreased and synthesis of HA by cumulus cells was reduced. Lapatinib (10 μM) inhibited FSH/LH-induced oocyte meiotic maturation. Progesterone production increased after OCC stimulation with FSH/LH and was significantly decreased by lapatinib (10 μM). CONCLUSION(S): Lapatinib inhibits oocyte maturation and reduces expression of cumulus expansion-associated transcripts, and synthesis of HA and P in OCC cultured in vitro in FSH/LH-supplemented medium.
- MeSH
- buněčná diferenciace účinky léků fyziologie MeSH
- chinazoliny farmakologie MeSH
- folikuly stimulující hormon farmakologie MeSH
- inhibitory růstu farmakologie MeSH
- kultivované buňky MeSH
- kumulární buňky cytologie účinky léků MeSH
- meióza účinky léků fyziologie MeSH
- oocyty cytologie účinky léků MeSH
- prasata MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- antitumorózní látky farmakologie chemie MeSH
- inhibitory enzymů farmakologie chemie MeSH
- inhibitory růstu farmakologie chemie MeSH
- jaderné proteiny MeSH
- kinasy CDC2-CDC28 antagonisté a inhibitory chemie MeSH
- nádorové buněčné linie MeSH
- protoonkogenní proteiny metabolismus MeSH
- puriny farmakologie chemie MeSH
- vazebná místa MeSH
- MeSH
- apoptóza účinky záření MeSH
- inhibitory růstu farmakologie MeSH
- membránové glykoproteiny farmakologie chemie MeSH
- myelodysplastické syndromy patologie MeSH
- myeloidní leukemie patologie MeSH
- myši MeSH
- nádorové kmenové buňky účinky léků MeSH
- tumor nekrotizující faktory farmakologie genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- MeSH
- buněčné dělení MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- folikulární tekutina chemie MeSH
- gelová chromatografie MeSH
- inhibitory růstu farmakologie chemie izolace a purifikace MeSH
- meióza MeSH
- oocyty cytologie MeSH
- skot MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- MeSH
- Crithidia analýza účinky léků metabolismus MeSH
- DNA biosyntéza MeSH
- inhibitory růstu izolace a purifikace farmakologie MeSH
- thymidin metabolismus MeSH
- tritium MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
