OBJECTIVE: By exposing mice carrying a deletion of NADPH oxidase isoform 4, NOX4, specifically in pancreatic β cells (βNOX4-/-) to nutrient excess stimulated by a high-fat diet (HFD), this study aimed to elucidate the role of β-cell redox status in the development of meta-inflammation within the diabetic phenotype. METHODS: The authors performed basic phenotyping of βNOX4-/- mice on HFD involving insulin and glycemic analyses, histochemistry of adipocytes, indirect calorimetry, and cytokine analyses. To characterize local inflammation, the study used caspase-1 activity assay, interleukin-1β immunochemistry, and real-time polymerase chain reaction during coculturing of β cells with macrophages. RESULTS: The phenotype of βNOX4-/- mice on HFD was not associated with hyperinsulinemia and hyperglycemia but showed accumulation of excessive lipids in epididymal fat and β cells. Surprisingly, mice showed significantly reduced systemic inflammation. Decreased interleukin-1β protein levels and downregulated NLRP3-inflammasome activity were observed on chronic glucose overload in βNOX4-/- isolated islets and NOX4-silenced INS1-E cells resulting in attenuated proinflammatory polarization of macrophages/monocytes in vitro and in situ and reduced local islet inflammation. CONCLUSIONS: Experimental evidence suggests that NOX4 pro-oxidant activity in β cells is involved in NLRP3-inflammasome activation during chronic nutrient overload and participates in local inflammatory signaling and perhaps toward peripheral tissues, contributing to a diabetic inflammatory phenotype.

• MeSH
• diabetes mellitus * MeSH
• inflamasomy metabolismus   MeSH
• interleukin-1beta metabolismus   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• NADPH-oxidasa 4 genetika   MeSH
• protein NLRP3 * metabolismus   MeSH
• zánět MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Effects of low-frequency electromagnetic fields (LF EMF) on the activation of different tissue recovery processes have already been fully understood. Preliminary recent data demonstrated that a special case of sinusoidal electromagnetic fields, known as amplitude-modulated currents (AMC) could have a potential to accelerate the cell metabolism or cell migration. An AMC generator was designed to generate sinusoidal induced electric currents with the amplitude modulation and the harmonic carrier frequency of 5,000 Hz was modulated by frequencies of 1 to 100 Hz. The magnetic field peak was 6 mT, electric field intensity 2 V/m and the current density of induced electrical currents was approximately 1 A/m(2). The coil of the generator was adapted to easy handling and safe integration into the shelf of the CO(2) incubator. The shelf with the coil was prepared for the introduction of cells in standard plastic in vitro chambers. The tests focused on cells with migratory capacity after injury or during immunological processes and thus, mesenchymal stromal cells (MSC), dendritic cells (DC), and fibroblasts were chosen. The tests involved exposures of the cells to LF EMF (180 min/day) every day, for a period of three days, before examining them for cell death, morphology changes, and CD markers. The samples were tested by using MTT assay and the effects on the intracellular concentration of reactive oxygen species were quantified. The cell migration was finally measured with the help of the transwell migration assay. None of the cell types showed any decrease in the cell viability after the LF EMF application and the cells displayed minimum changes in reactive oxygen species. Functional changes (acceleration of cell migration) after AMC exposure were statistically significant for the MSC samples only. The acceleration of MSCs is associated with the production of MMP by these cells. The EMF has a potential to be a safe, clinically applicable selective activator of MSC homing, MSC paracrine production, and subsequent regeneration processes.

• MeSH
• buňky 3T3 MeSH
• dendritické buňky fyziologie   MeSH
• elektromagnetická pole * MeSH
• fibroblasty fyziologie   MeSH
• lidé MeSH
• matrixová metaloproteinasa 2 metabolismus   MeSH
• mezenchymální kmenové buňky fyziologie   MeSH
• myši MeSH
• parakrinní signalizace MeSH
• pohyb buněk * MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

2,3-dehydrosilybin (DHS) is a minor component of silymarin, Silybum marianum seed extract, used in some dietary supplements. One of the most promising activities of this compound is its anticancer and cardioprotective activity that results, at least partially, from its cytoprotective, antioxidant, and chemopreventive properties. The present study investigated the cardioprotective effects of DHS in myocardial ischemia and reperfusion injury in rats. Isolated hearts were perfused by the Langendorff technique with low dose DHS (100 nM) prior to 30 min of ischemia induced by coronary artery occlusion. After 60 min of coronary reperfusion infarct size was determined by triphenyltetrazolium staining, while lactatedehydrogenase activity was evaluated in perfusate samples collected at several timepoints during the entire perfusion procedure. Signalosomes were isolated from a heart tissue after reperfusion and involved signalling proteins were detected. DHS reduced the extent of infarction compared with untreated control hearts at low concentration; infarct size as proportion of ischemic risk zone was 7.47 ± 3.1% for DHS versus 75.3 ± 4.8% for ischemia. This protective effect was comparable to infarct limitation induced by ischemic preconditioning (22.3 ± 4.5%). Selective inhibition of Src-family kinases with PP2 (4-Amino-3-(4-chlorophenyl)-1-(t-butyl)-1H-pyrazolo[3,4-d]pyrimidine) abrogated the protection afforded by DHS. This study provides experimental evidence that DHS can mediate Src-kinase-dependent cardioprotection against myocardial damage produced by ischemia/reperfusion injury.

• MeSH
• infarkt myokardu farmakoterapie   MeSH
• kardiotonika farmakologie   MeSH
• krysa rodu rattus MeSH
• potkani Wistar MeSH
• reperfuzní poškození farmakoterapie   MeSH
• silymarin farmakologie   MeSH
• srdce účinky léků   MeSH
• techniky in vitro MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The exposure of naked unprotected skin to solar radiation may result in numerous acute and chronic undesirable effects. Evidence suggests that silymarin, a standardized extract from Silybum marianum (L.) Gaertn. seeds, and its major component silybin suppress UVB-induced skin damage. Here, we aimed to investigate the UVA-protective effects of silymarin's less abundant flavonolignans, specifically isosilybin (ISB), silychristin (SC), silydianin (SD), and 2,3-dehydrosilybin (DHSB). Normal human dermal fibroblasts (NHDF) pre-treated for 1 h with flavonolignans were then exposed to UVA light using a solar simulator. Their effects on reactive oxygen species (ROS), carbonylated proteins and glutathione (GSH) level, caspase-3 activity, single-strand breaks' (SSBs) formation and protein level of matrix metalloproteinase-1 (MMP-1), heme oxygenase-1 (HO-1), and heat shock protein (HSP70) were evaluated. The most pronounced preventative potential was found for DHSB, a minor component of silymarin, and SC, the second most abundant flavonolignan in silymarin. They had significant effects on most of the studied parameters. Meanwhile, a photoprotective effect of SC was mostly found at double the concentration of DHSB. ISB and SD protected against GSH depletion, the generation of ROS, carbonylated proteins and SSBs, and caspase-3 activation, but had no significant effect on MMP-1, HO-1, or HSP70. In summary, DHSB and to a lesser extent other silymarin flavonolignans are potent UVA-protective compounds. However, due to the in vitro phototoxic potential of DHSB published elsewhere, further studies are needed to exclude phototoxicity for humans as well as to confirm our results on human skin ex vivo and in vivo.

• MeSH
• cytoprotekce účinky léků   MeSH
• fibroblasty účinky léků   účinky záření   MeSH
• glutathion metabolismus   MeSH
• hemoxygenasa-1 metabolismus   MeSH
• jednořetězcové zlomy DNA účinky záření   MeSH
• karbonylace proteinů účinky záření   MeSH
• kaspasa 3 metabolismus   MeSH
• kultivované buňky MeSH
• kůže účinky záření   MeSH
• lidé MeSH
• matrixová metaloproteinasa 1 metabolismus   MeSH
• přípravky chránící proti slunci farmakologie   MeSH
• proteiny tepelného šoku HSP70 metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• silymarin analogy a deriváty   farmakologie   MeSH
• ultrafialové záření škodlivé účinky   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Exposure to solar radiation is a major cause of environmental human skin damage. The main constituent of solar UV light is UVA radiation (320-400 nm); however, the need for protection against UVA has been marginalized for a long time. As a result, there is still a lack of useful agents for UVA protection. In this study, the effect of silymarin (SM) and its main constituent silybin (SB) pre-treatment on UVA-stimulated damage to primary human dermal fibroblasts were carried out. The cells were pre-treated for 1 h with SB or SM and then were exposed to UVA light, using a solar simulator. The effect of SB and SM on reactive oxygen species (ROS) and glutathione (GSH) level, caspase-3 activity, single-strand breaks (SSB) formation and protein level of matrix metalloproteinase-1 (MMP-1), heme oxygenase-1 (HO-1), and heat shock protein (HSP70) was evaluated. Treatment with both SM and SB resulted in a reduction in UVA-stimulated ROS generation and SSB production, as well as in the prevention of GSH depletion, a decrease in the activation of caspase-3 and protein level of MMP-1. They also moderately increased HO-1 level and reduced HSP70 level. Our data showed that both SM and SB are non-phototoxic and have UVA-photoprotective potential and could be useful agents for UV-protective dermatological preparations.

• MeSH
• fibroblasty účinky léků   patologie   účinky záření   MeSH
• glutathion metabolismus   MeSH
• hemoxygenasa-1 metabolismus   MeSH
• kaspasa 3 metabolismus   MeSH
• kultivované buňky MeSH
• kůže patologie   účinky záření   MeSH
• lidé MeSH
• matrixová metaloproteinasa 1 metabolismus   MeSH
• poškození DNA MeSH
• primární buněčná kultura MeSH
• proteiny tepelného šoku HSP70 metabolismus   MeSH
• radiační poranění farmakoterapie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• silibinin MeSH
• silymarin terapeutické užití   MeSH
• sluneční záření MeSH
• ultrafialové záření škodlivé účinky   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

2,3-dehydrosilybin (DHS) is a minor flavonolignan component of Silybum marianum seed extract known for its hepatoprotective activity. Recently we identified DHS as a potentially cardioprotective substance during hypoxia/reoxygenation in isolated neonatal rat cardiomyocytes. This is the first report of positive inotropic effect of DHS on perfused adult rat heart. When applied to perfused adult rat heart, DHS caused a dose-dependent inotropic effect resembling that of catecholamines. The effect was apparent with DHS concentration as low as 10 nM. Suspecting direct interaction with β-adrenergic receptors, we tested whether DHS can trigger β agonist-dependent gene transcription in a model cell line. While DHS alone was unable to trigger β agonist-dependent gene transcription, it enhanced the effect of isoproterenol, a known unspecific β agonist. Further tests confirmed that DHS could not induce cAMP accumulation in isolated neonatal rat cardiomyocytes even though high concentrations (≥ 10 μM) of DHS were capable of decreasing phosphodiesterase activity. Pre-treatment of rats with reserpine, an indole alkaloid which depletes catecholamines from peripheral sympathetic nerve endings, abolished the DHS inotropic effect in perfused hearts. Our data suggest that DHS causes the inotropic effect without acting as a β agonist. Hence we identify DHS as a novel inotropic agent.

• MeSH
• buněčné linie MeSH
• kardiomyocyty fyziologie   MeSH
• kardiotonika farmakologie   MeSH
• kontrakce myokardu účinky léků   MeSH
• krysa rodu rattus MeSH
• ostropestřec mariánský MeSH
• potkani Wistar MeSH
• reserpin farmakologie   MeSH
• rostlinné přípravky farmakologie   MeSH
• silymarin farmakologie   MeSH
• srdce účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Ischemic postconditioning and remote conditioning are potentially useful tools for protecting ischemic myocardium. This study tested the hypothesis that 2,3-dehydrosilybin (DHS), a flavonolignan component of Silybum marianum, could attenuate cardiomyocyte damage following hypoxia/reoxygenation by decreasing the generation of reactive oxygen species (ROS). After 5-6 days of cell culture in normoxic conditions the rat neonatal cardiomyocytes were divided into four groups. Control group (9 h at normoxic conditions), hypoxia/reoxygenation group (3 h at 1 % O₂, 94 % N₂and 5 % CO₂followed by 10 min of 10 micromol·l⁻¹DHS and 6 h of reoxygenation in normoxia) and postconditioning group (3 h of hypoxia, three cycles of 5 min reoxygenation and 5 min hypoxia followed by 6 h of normoxia). Cell viability assessed by propidium iodide staining was decreased after DHS treatment consistent with increased levels of lactatedehydrogenase (LDH) after reoxygenation. LDH leakage was significantly reduced when cardiomyocytes in the H/Re group were exposed to DHS. DHS treatment reduced H₂O₂production and also decreased the generation of ROS in the H/Re group as evidenced by a fluorescence indicator. DHS treatment reduces reoxygenation-induced injury in cardiomyocytes by attenuation of ROS generation, H₂O₂and protein carbonyls levels. In addition, we found that both the postconditioning protocol and the DHS treatment are associated with restored ratio of phosphorylated/total protein kinase C epsilon, relative to the H/Re group. In conclusion, our data support the protective role of DHS in hypoxia/reperfusion injury and indicate that DHS may act as a postconditioning mimic.

• MeSH
• antioxidancia farmakologie   MeSH
• cytoprotekce MeSH
• fosforylace MeSH
• ischemické přivykání MeSH
• karbonylace proteinů účinky léků   MeSH
• kardiomyocyty účinky léků   metabolismus   patologie   MeSH
• kultivované buňky MeSH
• L-laktátdehydrogenasa metabolismus   MeSH
• novorozená zvířata MeSH
• oxidační stres účinky léků   MeSH
• peroxid vodíku metabolismus   MeSH
• potkani Wistar MeSH
• proteinkinasa C-epsilon metabolismus   MeSH
• reperfuzní poškození myokardu metabolismus   patologie   prevence a kontrola   MeSH
• signální transdukce účinky léků   MeSH
• silymarin farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

OBJECTIVE: Cell therapies have emerged as a promising approach in medicine. The basis of each therapy is the injection of 1-100×10(6) cells with regenerative potential into some part of the body. Mesenchymal stromal cells (MSCs) are the most used cell type in the cell therapy nowadays, but no gold standard for the labeling of the MSCs for magnetic resonance imaging (MRI) is available yet. This work evaluates our newly synthesized uncoated superparamagnetic maghemite nanoparticles (surface-active maghemite nanoparticles - SAMNs) as an MRI contrast intracellular probe usable in a clinical 1.5 T MRI system. METHODS: MSCs from rat and human donors were isolated, and then incubated at different concentrations (10-200 μg/mL) of SAMN maghemite nanoparticles for 48 hours. Viability, proliferation, and nanoparticle uptake efficiency were tested (using fluorescence microscopy, xCELLigence analysis, atomic absorption spectroscopy, and advanced microscopy techniques). Migration capacity, cluster of differentiation markers, effect of nanoparticles on long-term viability, contrast properties in MRI, and cocultivation of labeled cells with myocytes were also studied. RESULTS: SAMNs do not affect MSC viability if the concentration does not exceed 100 μg ferumoxide/mL, and this concentration does not alter their cell phenotype and long-term proliferation profile. After 48 hours of incubation, MSCs labeled with SAMNs show more than double the amount of iron per cell compared to Resovist-labeled cells, which correlates well with the better contrast properties of the SAMN cell sample in T2-weighted MRI. SAMN-labeled MSCs display strong adherence and excellent elasticity in a beating myocyte culture for a minimum of 7 days. CONCLUSION: Detailed in vitro tests and phantom tests on ex vivo tissue show that the new SAMNs are efficient MRI contrast agent probes with exclusive intracellular uptake and high biological safety.

• MeSH
• buněčný tracking metody   MeSH
• dextrany chemie   farmakokinetika   toxicita   MeSH
• fyziologie buňky účinky léků   MeSH
• kontrastní látky chemie   farmakokinetika   toxicita   MeSH
• krysa rodu rattus MeSH
• kultivované buňky MeSH
• lidé MeSH
• magnetická rezonanční tomografie MeSH
• magnetické nanočástice chemie   toxicita   MeSH
• mezenchymální kmenové buňky chemie   cytologie   účinky léků   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Silybin and its congeners belong to a group of flavonolignans with strong biological activities. These compounds are potentially applicable in human medicine, e. g. due to their cytoprotective activity. As a part of herbal preparations available on the open market, they face the risk of potential negative drug-drug interactions. This review aims to evaluate current knowledge on the metabolism of these compounds by biotransformation enzymes, interactions with other drugs, their pharmacokinetics, and bioavailability. While silybin and its derivatives interact with cytochrome P450s, only metabolism of silybin by cytochrome P450 2C8 poses a risk of adverse effects. The main biotransformation route of silybin and derivatives was identified as conjugation, which is stereospecific in case of silybin. Studies of the metabolism, pharmacokinetics, potentional drug--drug interactions and increasing bioavailability of these flavonolignans play an important facet of possible therapeutical use of these compounds. The goal of our review is to aid future developments in the area of silybin research.

• MeSH
• antioxidancia chemie   metabolismus   farmakokinetika   MeSH
• biologická dostupnost MeSH
• biotransformace MeSH
• glukuronidy metabolismus   MeSH
• lidé MeSH
• silymarin analogy a deriváty   chemie   metabolismus   farmakokinetika   MeSH
• stereoizomerie MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

Reactive oxygen species (ROS) originating from mitochondria are perceived as a factor contributing to cell aging and means have been sought to attenuate ROS formation with the aim of extending the cell lifespan. Silybin and dehydrosilybin, two polyphenolic compounds, display a plethora of biological effects generally ascribed to their known antioxidant capacity. When investigating the cytoprotective effects of these two compounds in the primary cell cultures of neonatal rat cardiomyocytes, we noted the ability of dehydrosilybin to de-energize the cells by monitoring JC-1 fluorescence. Experiments evaluating oxygen consumption and membrane potential revealed that dehydrosilybin uncouples the respiration of isolated rat heart mitochondria albeit with a much lower potency than synthetic uncouplers. Furthermore, dehydrosilybin revealed a very high potency in suppressing ROS formation in isolated rat heart mitochondria with IC(50) = 0.15 μM. It is far more effective than its effect in a purely chemical system generating superoxide or in cells capable of oxidative burst, where the IC(50) for dehydrosilybin exceeds 50 μM. Dehydrosilybin also attenuated ROS formation caused by rotenone in the primary cultures of neonatal rat cardiomyocytes. We infer that the apparent uncoupler-like activity of dehydrosilybin is the basis of its ROS modulation effect in neonatal rat cardiomyocytes and leads us to propose a hypothesis on natural ischemia preconditioning by dietary polyphenols.

• MeSH
• analýza rozptylu MeSH
• benzimidazoly MeSH
• fluorescenční barviva MeSH
• inhibiční koncentrace 50 MeSH
• karbocyaniny MeSH
• kardiomyocyty metabolismus   MeSH
• krysa rodu rattus MeSH
• mitochondrie metabolismus   MeSH
• molekulární struktura MeSH
• potkani Wistar MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• rotenon toxicita   MeSH
• silymarin chemie   farmakologie   MeSH
• spotřeba kyslíku účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH