12-O-Tetradecanoylphorbol-13-acetate (TPA) is the most widely used diacylglycerol (DAG) mimetic agent and inducer of protein kinase C (PKC)-mediated cellular response in biomedical studies. TPA has been proposed as a pluripotent cell differentiation factor, but results obtained have been inconsistent. In the present study we show that TPA can be applied as a cardiomyogenesis-promoting factor for the differentiation of mouse embryonic stem (mES) cells in vitro. The mechanism of TPA action is mediated by the induction of extracellular signal-regulated kinase (ERK) activity and the subsequent phosphorylation of GATA4 transcription factor. Interestingly, general mitogens (FGF, EGF, VEGF and serum) or canonical WNT signalling did not mimic the effect of TPA. Moreover, on the basis of our results, we postulate that a TPA-sensitive population of cardiac progenitor cells exists at a certain time point (after days 6-8 of the differentiation protocol) and that the proposed treatment can be used to increase the multiplication of ES cell-derived cardiomyocytes.

• MeSH
• buněčná diferenciace účinky léků   MeSH
• embryonální kmenové buňky cytologie   účinky léků   metabolismus   MeSH
• extracelulárním signálem regulované MAP kinasy metabolismus   MeSH
• fosforylace MeSH
• kardiomyocyty cytologie   metabolismus   MeSH
• myši MeSH
• proteinkinasa C metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• tetradekanoylforbolacetát farmakologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• extracelulárním signálem regulované MAP kinasy MeSH
• proteinkinasa C MeSH
• tetradekanoylforbolacetát MeSH

To specify the site of action of the synthetic coumarin derivatives 7-hydroxy-3-(4'-hydroxyphenyl) coumarin (HHC) and 7-hydroxy-3-(4'-hydroxyphenyl) dihydrocoumarin (HHDC), we evaluated their effects on extra- and intracellular reactive oxygen species (ROS) formation in phorbol-myristate-13-acetate (PMA) stimulated human neutrophils. We studied also the effects of HHC and HHDC on possible molecular mechanisms which participate in the activation of NADPH oxidase, that is, on PKC activity, on phosphorylation of some PKC isoforms (α, βII, and δ), and on phosphorylation of the NADPH oxidase subunit p40(phox). Without affecting cytotoxicity, both coumarines tested were effective inhibitors/scavengers of ROS produced by neutrophils on extracellular level. HHC markedly diminished oxidant production and also, intracellularly, decreased PKC activity and partly phosphorylation of PKCα, βII. On the other hand, we did not observe any effect of coumarin derivatives on phosphorylation of PKC δ and on phosphorylation of the NADPH oxidase subunit p40(phox), which were suggested to be involved in the PMA-dependent intracellular activation process. In agreement with our previous findings, we assume that the different molecular structures of HHC and HHDC with their different physicochemical and free radical scavenging characteristics are responsible for their diverse effects on the parameters tested.

• MeSH
• adenosintrifosfát metabolismus   MeSH
• aktivace neutrofilů účinky léků   MeSH
• bezbuněčný systém MeSH
• buněčná smrt účinky léků   MeSH
• dospělí MeSH
• extracelulární prostor účinky léků   metabolismus   MeSH
• fosfoproteiny metabolismus   MeSH
• fosforylace účinky léků   MeSH
• inhibiční koncentrace 50 MeSH
• intracelulární prostor účinky léků   metabolismus   MeSH
• izoenzymy metabolismus   MeSH
• kinetika MeSH
• kumariny chemie   farmakologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• luminiscenční měření MeSH
• mladý dospělý MeSH
• neutrofily cytologie   účinky léků   enzymologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• tetradekanoylforbolacetát farmakologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adenosintrifosfát MeSH
• coumarin MeSH Prohlížeč
• fosfoproteiny MeSH
• izoenzymy MeSH
• kumariny MeSH
• neutrophil cytosol factor 40K MeSH Prohlížeč
• reaktivní formy kyslíku MeSH
• tetradekanoylforbolacetát MeSH