Glucagon-like peptide-1 receptor (GLP1R) agonist is an incretin hormone and regulates glucose metabolism. However, phthalates, known as endocrine disruptors, can interfere with hormone homeostasis. In the present study, we aimed to estimate the impact of GLP1R agonist on di(2 ethylhexyl) phthalate (DEHP)-induced atherosclerosis. For this purpose, the effects of GLP1R agonist on various atherogenesis-related cellular processes and pathways were assessed in vascular smooth muscle cells (VSMCs). DEHP-induced cell proliferation and migration were significantly decreased by GLP1R agonist in VSMCs. Protein levels of matrix metalloproteinase (MMP)-2 and MMP-9 were significantly decreased in cells exposed to GLP1R agonist, compared with DEHP-treated cells. Expression levels of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 were also reduced in GLP1R agonist-treated cells. Similarly, DEHP-associated phosphorylation of protein kinase B and extracellular signal-regulated kinase 1/2 was decreased in GLP1R agonist-treated cells, compared with DEHP-treated cells. Our findings suggest that treatment with GLP1R agonist counteracts the activation of pathways related to atherosclerosis.

• MeSH
• ateroskleróza chemicky indukované   farmakoterapie   metabolismus   patologie   MeSH
• diethylhexylftalát toxicita   MeSH
• inkretiny farmakologie   MeSH
• krysa rodu rattus MeSH
• kultivované buňky MeSH
• mezibuněčná adhezivní molekula-1 genetika   metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 3 genetika   metabolismus   MeSH
• proliferace buněk MeSH
• receptor pro glukagonu podobný peptid 1 agonisté   MeSH
• signální transdukce účinky léků   MeSH
• svaly hladké cévní účinky léků   metabolismus   patologie   MeSH
• změkčovadla toxicita   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Cyanobacterial toxin cylindrospermopsin (CYN) is an emerging freshwater contaminant, whose expanding environmental occurrence might result into increased human health risks. CYN is potent hepatotoxin, with cytotoxicity and genotoxicity documented in primary hepatocytes or hepatoma cell lines. However, there is only limited information about CYN effects on adult human liver stem cells (LSCs), which play an important role in liver tissue development, regeneration and repair. In our study with human liver cell line HL1-hT1 which expresses characteristics of LSCs, CYN was found to be cytotoxic and increasing cell death after 24-48 h exposure to concentrations >1 μM. Subcytotoxic 1 μM concentration did not induce cell death or membrane damage, but inhibited cellular processes related to energy production, leading to a growth stagnation after >72 h. Interestingly, these effects were not associated with increased DNA damage, reactive oxygen species production, or endoplasmic reticulum stress. However, CYN induced a sustained (24-48 h) activation of mitogen-activated protein kinases ERK1/2 and p38, and increased expression of stress-related transcription factor ATF3. Thus, LSCs were not primarily affected by CYN-induced genotoxicity and oxidative stress, but via activation of signaling and transcriptional pathways critical for regulation of cell proliferation, stress responses, cell survival and inflammation. Alterations of LSCs during CYN-induced liver injury, including the role of nongenotoxic mechanisms, should be therefore considered in mechanistic assessments of chronic CYN hepatotoxicity and hepatocarcinogenicity.

• MeSH
• bakteriální toxiny toxicita   MeSH
• buněčné linie MeSH
• hepatocyty účinky léků   MeSH
• játra metabolismus   MeSH
• kmenové buňky MeSH
• lidé MeSH
• MAP kinasový signální systém MeSH
• mikrocystiny MeSH
• mitogenem aktivovaná proteinkinasa 1 metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 3 metabolismus   MeSH
• mitogenem aktivované proteinkinasy p38 metabolismus   MeSH
• mořské toxiny MeSH
• oxidační stres účinky léků   MeSH
• poškození DNA MeSH
• proliferace buněk MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• testy toxicity MeSH
• uracil analogy a deriváty   toxicita   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The maturation of mammalian oocytes in vitro can be stimulated by gonadotropins (follicle-stimulating hormone, FSH) or their intrafollicular mediator, epidermal growth factor (EGF)-like peptide-amphiregulin (AREG). We have shown previously that in pig cumulus-oocyte complexes (COCs), FSH induces expression and the synthesis of AREG that binds to EGF receptor (EGFR) and activates the mitogen-activated protein kinase 3/1 (MAPK3/1) signaling pathway. However, in this study we found that FSH also caused a rapid activation of MAPK3/1 in the cumulus cells, which cannot be explained by the de novo synthesis of AREG. The rapid MAPK3/1 activation required EGFR tyrosine kinase (TK) activity, was sensitive to SRC proto-oncogene non-receptor tyrosine kinase (SRC)-family and protein kinase C (PKC) inhibitors, and was resistant to inhibitors of protein kinase A (PKA) and metalloproteinases. AREG also induced the rapid activation of MAPK3/1 in cumulus cells, but this activation was only dependent on the EGFR TK activity. We conclude that in cumulus cells, FSH induces a rapid activation of MAPK3/1 by the ligand-independent transactivation of EGFR, requiring SRC and PKC activities. This rapid activation of MAPK3/1 precedes the second mechanism participating in the generation and maintenance of active MAPK3/1-the ligand-dependent activation of EGFR depending on the synthesis of EGF-like peptides.

• MeSH
• aktivace transkripce MeSH
• amfiregulin metabolismus   MeSH
• erbB receptory metabolismus   MeSH
• extracelulárním signálem regulované MAP kinasy genetika   MeSH
• folikuly stimulující hormon farmakologie   MeSH
• IVM techniky MeSH
• kultivované buňky MeSH
• kumulární buňky cytologie   účinky léků   metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 1 genetika   MeSH
• mitogenem aktivovaná proteinkinasa 3 genetika   MeSH
• oocyty cytologie   účinky léků   metabolismus   MeSH
• prasata MeSH
• signální transdukce účinky léků   MeSH
• skupina kinas odvozených od src-genu metabolismus   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The circadian clock in the suprachiasmatic nucleus (SCN) regulates daily rhythms in physiology and behaviour and is an important part of the mammalian homeostatic system. Previously, we have shown that systemic inflammatory stimulation with lipopolysaccharide (LPS) induced the daytime-dependent phosphorylation of STAT3 in the SCN. Here, we demonstrate the LPS-induced Stat3 mRNA expression in the SCN and show also the circadian rhythm in Stat3 expression in the SCN, with high levels during the day. Moreover, we examined the effects of LPS (1mg/kg), applied either during the day or the night, on the rhythm in locomotor activity of male Wistar rats. We observed that recovery of normal locomotor activity patterns took longer when the animals were injected during the night. The clock genes Per1, Per2 and Nr1d1, and phosphorylation of kinases ERK1/2 and GSK3β are sensitive to external cues and function as the molecular entry for external signals into the circadian clockwork. We also studied the immediate changes in these clock genes expressions and the phosphorylation of ERK1/2 and GSK3β in the suprachiasmatic nucleus in response to daytime or night-time inflammatory stimulation. We revealed mild and transient changes with respect to the controls. Our data stress the role of STAT3 in the circadian clock response to the LPS and provide further evidence of the interaction between the circadian clock and immune system.

• MeSH
• cirkadiánní rytmus účinky léků   MeSH
• krysa rodu rattus MeSH
• lipopolysacharidy toxicita   MeSH
• lokomoce účinky léků   MeSH
• MAP kinasový signální systém účinky léků   MeSH
• mitogenem aktivovaná proteinkinasa 3 metabolismus   MeSH
• nucleus suprachiasmaticus metabolismus   patologie   MeSH
• potkani Wistar MeSH
• regulace genové exprese účinky léků   MeSH
• transkripční faktor STAT3 biosyntéza   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Although the involvement of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) pathway in the regulation of cytostatic factor (CSF) activity; as well as in microtubules organization during meiotic maturation of oocytes; has already been described in detail; rather less attention has been paid to the role of ERK1/2 in the regulation of mRNA translation. However; important data on the role of ERK1/2 in translation during oocyte meiosis have been documented. This review focuses on recent findings regarding the regulation of translation and the role of ERK1/2 in this process in the meiotic cycle of mammalian oocytes. The specific role of ERK1/2 in the regulation of mammalian target of rapamycin (mTOR); eukaryotic translation initiation factor 4E (eIF4E) and cytoplasmic polyadenylation element binding protein 1 (CPEB1) activity is addressed along with additional focus on the other key players involved in protein translation.

• MeSH
• cytoplazma genetika   metabolismus   MeSH
• eukaryotický iniciační faktor 4E metabolismus   MeSH
• faktory štěpení a polyadenylace mRNA metabolismus   MeSH
• fosfatidylinositol-3-kinasy metabolismus   MeSH
• lidé MeSH
• meióza * MeSH
• messenger RNA genetika   metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 1 metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 3 metabolismus   MeSH
• mitogenem aktivované proteinkinasy metabolismus   MeSH
• oocyty metabolismus   MeSH
• polyadenylace MeSH
• proteosyntéza * MeSH
• signální transdukce MeSH
• TOR serin-threoninkinasy metabolismus   MeSH
• vazba proteinů MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

The transient receptor potential vanilloid 3 (TRPV3) channel is a Ca2+-permeable thermosensitive ion channel widely expressed in keratinocytes, where together with epidermal growth factor receptor (EGFR) forms a signaling complex regulating epidermal homeostasis. Proper signaling through this complex is achieved and maintained via several pathways in which TRPV3 activation is absolutely required. Results of recent studies have suggested that low-level constitutive activity of TRPV3 induces EGFR-dependent signaling that, in turn, amplifies TRPV3 via activation of the mitogen-activated protein kinase ERK in a positive feedback loop. Here, we explored the molecular mechanism that increases TRPV3 activity through EGFR activation. We used mutagenesis and whole-cell patch clamp experiments on TRPV3 channels endogenously expressed in an immortalized human keratinocyte cell line (HaCaT) and in transiently transfected HEK293T cells and found that the sensitizing effect of EGFR on TRPV3 is mediated by ERK. We observed that ERK-mediated phosphorylation of TRPV3 alters its responsiveness to repeated chemical stimuli. Among several putative ERK phosphorylation sites, we identified threonine 264 in the N-terminal ankyrin repeat domain as the most critical site for the ERK-dependent modulation of TRPV3 channel activity. Of note, Thr264 is in close vicinity to a structurally and functionally important TRPV3 region comprising an atypical finger 3 and oxygen-dependent hydroxylation site. In summary, our findings indicate that Thr264 in TRPV3 is a key ERK phosphorylation site mediating EGFR-induced sensitization of the channel to stimulate signaling pathways involved in regulating skin homeostasis.

• MeSH
• epidermální růstový faktor metabolismus   MeSH
• erbB receptory agonisté   metabolismus   MeSH
• fosforylace účinky léků   MeSH
• HEK293 buňky MeSH
• interakční proteinové domény a motivy MeSH
• kationtové kanály TRPV agonisté   chemie   genetika   metabolismus   MeSH
• keratinocyty účinky léků   enzymologie   metabolismus   MeSH
• lidé MeSH
• MAP kinasový signální systém * účinky léků   MeSH
• metoda terčíkového zámku MeSH
• mitogenem aktivovaná proteinkinasa 3 chemie   genetika   metabolismus   MeSH
• modulátory membránového transportu farmakologie   MeSH
• monoterpeny farmakologie   MeSH
• mutace MeSH
• mutageneze cílená MeSH
• posttranslační úpravy proteinů účinky léků   MeSH
• rekombinantní proteiny chemie   metabolismus   MeSH
• sloučeniny boru farmakologie   MeSH
• threonin metabolismus   MeSH
• transformované buněčné linie MeSH
• upregulace * účinky léků   MeSH
• zelené fluorescenční proteiny genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The non-dioxin-like environmental toxicant 2,2',4,4',5,5'-hexachlorobiphenyl (PCB153), member of a group of persistent organic pollutants wide-spread throughout the environment, reduces gap junction intercellular communication (GJIC), an event possibly associated with tumor promotion. Since very few studies have investigated the signaling effectors and mode(s) of action of PCB153, and it is known that the gap junction (GJ) protein Cx43 can be regulated by the bioactive sphingolipid (SL) sphingosine 1-phosphate (S1P), this in vitro study mainly addresses whether SL metabolism is affected by PCB153 in rat liver epithelial WB-F344 cells. PCB153 treatment obtained significant changes in the S1P/ceramide (Cer) ratio, known to be crucial in determining cell fate. In particular, an increase in S1P at 30 min and a decrease of the bioactive lipid at 3 h were observed, whereas Cer level increased at 1 h and 24 h. Notably, a time-dependent modulation of sphingosine kinase (SphK), the enzyme responsible for S1P synthesis, and of its regulators, ERK1/2 and protein phosphatase PP2A, supports the involvement of these signaling effectors in PCB153 toxicity. Electrophysiological analyses, furthermore, indicated that the lipophilic environmental toxicant significantly reduced GJ biophysical properties, affecting both voltage-dependent (such as those formed by Cx43 and/or Cx32) and voltage-independent channels, thereby demonstrating that PCB153 may act differently on GJs formed by distinct Cx isoforms. SphK down-regulation alone induced GJIC impairment, and, when combined with PCB153, the acute effect on GJ suppression was additive. Moreover, after enzyme-specific gene silencing, the SphK1 isoform appears to be responsible for down-regulating Cx43 expression, while being the target of PCB153 at short-term exposure. In conclusion, we provide the first evidence of novel effectors in PCB153 toxic action in rat liver stem-like cells, leading us to consider SLs as potential markers for preventing GJIC deregulation and, thus, the tumorigenic action elicited by this environmental toxicant.

• MeSH
• dioxiny toxicita   MeSH
• elektrofyziologie metody   MeSH
• fosfotransferasy s alkoholovou skupinou jako akceptorem antagonisté a inhibitory   genetika   metabolismus   MeSH
• játra cytologie   účinky léků   MeSH
• konexin 43 metabolismus   MeSH
• krysa rodu rattus MeSH
• kultivované buňky MeSH
• lysofosfolipidy metabolismus   MeSH
• mezerový spoj účinky léků   fyziologie   MeSH
• mitogenem aktivovaná proteinkinasa 3 metabolismus   MeSH
• polychlorované bifenyly toxicita   MeSH
• proteinfosfatasa 2 genetika   metabolismus   MeSH
• sfingolipidy metabolismus   MeSH
• sfingosin analogy a deriváty   metabolismus   MeSH
• signální transdukce účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The production of prostaglandin E2 (PGE2) seems to play an important role in the ovulation process. PGE2 was found to induce cumulus expansion and meiosis resumption in mice, but little is known about its role in pigs. The goals of this study were (a) to assess the effect of PGE2 on the expression levels of cumulus expansion-related genes, (b) to define the signaling pathways that drive the PGE2-stimulated expression of cumulus expansion-related genes, (c) to measure the effect of PGE2 on the activation of key signaling molecules (MAPK3/1, PKB) and on hyaluronan production in cumulus cells, and (d) to assess the effect of PGE2 on meiosis resumption. We documented that PGE2 is able to induce the expression of cumulus expansion-related genes (HAS2, TNFAIP6) as well as genes involved in steroidogenesis (CYP11A1) or prostaglandin production (PTGS2). PGE2 is able to activate PKB and MAPK3/1 and induce mild cumulus expansion and meiosis resumption, but less efficiently than FSH.

• MeSH
• aktivace enzymů účinky léků   MeSH
• dinoproston farmakologie   MeSH
• down regulace účinky léků   MeSH
• kumulární buňky cytologie   účinky léků   metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 1 metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 3 metabolismus   MeSH
• oocyty cytologie   účinky léků   MeSH
• prasata MeSH
• protoonkogenní proteiny c-akt metabolismus   MeSH
• regulace genové exprese účinky léků   MeSH
• upregulace účinky léků   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Gold alkynyl complexes with phosphane ligands of the type (alkynyl)Au(I)(phosphane) represent a group of bioorganometallics, which has only recently been evaluated biologically in more detail. Structure-activity-relationship studies regarding the residues of the phosphane ligand (P(Ph)3, P(2-furyl)3, P(DAPTA)3, P(PTA)3, P(Et)3, P(Me)3) of complexes with an 4-ethynylanisole alkyne ligand revealed no strong differences concerning cytotoxicity. However, a relevant preference for the heteroatom free alkyl/aryl residues concerning inhibition of the target enzyme thioredoxin reductase was evident. Complex 1 with the triphenylphosphane ligand was selected for further studies, in which clear effects on cell morphology were monitored by time-lapse microscopy. Effects on cellular signaling were determined by ELISA microarrays and showed a significant induction of the phosphorylation of ERK1 (extracellular signal related kinase 1), ERK2 and HSP27 (heat shock protein 27) in HT-29 cells. Application of 1 in-vivo in a mouse xenograft model was found to be challenging due to the low solubility of the complex and required a formulation strategy based on a peanut oil nanoemulsion.

• MeSH
• anisoly chemie   MeSH
• buňky HT-29 MeSH
• fosfiny chemie   MeSH
• kationty jednomocné MeSH
• komplexní sloučeniny chemická syntéza   farmakologie   MeSH
• lidé MeSH
• ligandy MeSH
• mitogenem aktivovaná proteinkinasa 1 genetika   metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 3 genetika   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši nahé MeSH
• myši MeSH
• nádory plic farmakoterapie   patologie   MeSH
• organické sloučeniny zlata chemická syntéza   farmakologie   MeSH
• proliferace buněk účinky léků   MeSH
• proteiny tepelného šoku HSP27 genetika   metabolismus   MeSH
• protinádorové látky chemická syntéza   farmakologie   MeSH
• thioredoxin-disulfidreduktasa antagonisté a inhibitory   genetika   metabolismus   MeSH
• viabilita buněk účinky léků   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• xenogenní modely - testy protinádorové aktivity MeSH
• zlato chemie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

STUDY HYPOTHESIS: In women with preterm premature rupture of the membranes (PPROM), increased oxidative stress may accelerate premature cellular senescence, senescence-associated inflammation and proteolysis, which may predispose them to rupture. STUDY FINDING: We demonstrate mechanistic differences between preterm birth (PTB) and PPROM by revealing differences in fetal membrane redox status, oxidative stress-induced damage, distinct signaling pathways and senescence activation. WHAT IS KNOWN ALREADY: Oxidative stress-associated fetal membrane damage and cell cycle arrest determine adverse pregnancy outcomes, such as spontaneous PTB and PPROM. STUDY DESIGN, SAMPLES/MATERIALS, METHODS: Fetal membranes and amniotic fluid samples were collected from women with PTB and PPROM. Molecular, biochemical and histologic markers were used to document differences in oxidative stress and antioxidant enzyme status, DNA damage, secondary signaling activation by Ras-GTPase and mitogen-activated protein kinases, and activation of senescence between membranes from the two groups. MAIN RESULTS AND THE ROLE OF CHANCE: Oxidative stress was higher and antioxidant enzymes were lower in PPROM compared with PTB. PTB membranes had minimal DNA damage and showed activation of Ras-GTPase and ERK/JNK signaling pathway with minimal signs of senescence. PPROM had higher numbers of cells with DNA damage, prosenescence stress kinase (p38 MAPK) activation and signs of senescence. LIMITATIONS, REASONS FOR CAUTION: Samples were obtained retrospectively after delivery. The markers of senescence that we tested are specific but are not sufficient to confirm senescence as the pathology in PPROM. WIDER IMPLICATIONS OF THE FINDINGS: Oxidative stress-induced DNA damage and senescence are characteristics of fetal membranes from PPROM, compared with PTB with intact membranes. PTB and PPROM arise from distinct pathophysiologic pathways. Oxidative stress and oxidative stress-induced cellular damages are likely determinants of the mechanistic signaling pathways and phenotypic outcome. STUDY FUNDING AND COMPETING INTERESTS: This study is supported by developmental funds to Dr R. Menon from the Department of Obstetrics and Gynecology at The University of Texas Medical Branch at Galveston and funds to Dr M. Kacerovský from the Ministry of Health Czech Republic (UHHK, 001799906). The authors report no conflict of interest.

• MeSH
• dospělí MeSH
• extraembryonální obaly zranění   metabolismus   MeSH
• homeodoménové proteiny genetika   metabolismus   MeSH
• lamin typ B genetika   metabolismus   MeSH
• lidé MeSH
• MAP kinasa-kinasa 4 genetika   metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 1 genetika   metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 3 genetika   metabolismus   MeSH
• mitogenem aktivované proteinkinasy p38 genetika   metabolismus   MeSH
• novorozenec nedonošený MeSH
• novorozenec MeSH
• oxidační stres genetika   MeSH
• poškození DNA MeSH
• předčasný odtok plodové vody genetika   metabolismus   patologie   MeSH
• předčasný porod MeSH
• regulace genové exprese MeSH
• signální transdukce genetika   MeSH
• stanovení celkové genové exprese MeSH
• stárnutí buněk MeSH
• superoxiddismutasa genetika   metabolismus   MeSH
• těhotenství MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• novorozenec MeSH
• těhotenství MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH