Although hypothermic treatment has been reported to have some beneficial effects on ischaemia at the clinical level, the mechanism of ischaemia suppression by hypothermia remains unclear due to a lack of mechanism understanding and insufficient data. The aim of this study was to isolate and characterize microRNAs specifically expressed in ischaemia-hypothermia for the dihydropyrimidinase-like 3 (Dpysl3) gene. PC12 cells were induced with CoCl2 for chemical ischaemia and incubated at 32 °C for hypothermia. In ischaemia-hypothermia, four types of microRNAs (miR-106b-5p, miR-194-5p, miR-326-5p, and miR-497-5p) were highly related to the Dpysl3 gene based on exosomal microRNA analysis. Dpysl3 gene expression was up-regulated by miR-497-5p but down-regulated by miR-106b-5p, miR-194-5p and miR-326-5p. Our results suggest that these four microRNAs are involved in the regulation of Dpysl3 gene expression. These findings provide valuable clues that exosomal microRNAs could be used as therapeutic targets for effective treatment of ischaemia.

• MeSH
• buňky PC12 MeSH
• exprese genu MeSH
• hypotermie * genetika   MeSH
• ischemie MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• mikro RNA * genetika   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

CART (cocaine- and amphetamine-regulated transcript) peptides have been studied for ten years. We report specific binding of 125I-CART(61-102) to the rat adrenal pheochromocytoma PC12 cell line, both intact cells and cell membranes. Saturation binding to intact plated cells resulted in Kd of 0.48+/-0.16 nM and Bmax of 2228+/-529 binding sites/cell. 125I-CART(61-102) was also bound to PC12 cells differentiated using nerve growth factor to the neuronal phenotype with non-specific binding below 20%, and Kd of 1.90+/-0.27 nM and Bmax of 11,194+/-261 binding sites/cell. In competitive binding experiments, CART(61-102), CART(55-102) and di-iodinated CART(61-102) were bound to PC12 cell membranes with Ki in low nM range; their affinity to intact non-differentiated and differentiated cells was in low 10(-8) M range. In order to prove that iodination did not eliminate the pharmacological properties of CART, we tested the biological activity of di-iodinated CART(61-102). It decreased food intake in in vivo feeding experiment on fasted mice in a dose of 1 microg/mouse to the same extent as CART(61-102) in a dose of 0.5 microg/mouse.

• MeSH
• biologické modely MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčná membrána metabolismus   MeSH
• buňky PC12 MeSH
• časové faktory MeSH
• fenotyp MeSH
• feochromocytom MeSH
• financování organizované MeSH
• izotopy jodu metabolismus   MeSH
• kinetika MeSH
• krysa rodu rattus MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nádory nadledvin MeSH
• neurony metabolismus   patologie   MeSH
• neurotrofní faktory farmakologie   MeSH
• peptidové fragmenty metabolismus   MeSH
• přijímání potravy účinky léků   MeSH
• proteiny nervové tkáně farmakologie   metabolismus   MeSH
• radioizotopy jodu metabolismus   MeSH
• vazba proteinů MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH

CART (cocaine- and amphetamine-regulated transcript) peptide is a neuropeptide with a powerful central anorexigenic effect. Specific CART peptide binding sites, most likely CART peptide receptors, have been found in PC12 cells. This study further characterizes the CART peptide binding sites in PC12 cells. After differentiation to a neuronal phenotype with nerve growth factor, the number of CART peptide binding sites in PC12 cells tripled. Following dexamethasone treatment, which transforms PC12 cells into chromaffin-like cells, the number of CART peptide binding sites substantially decreased. CART peptide did not affect the differentiation or acetylcholinesterase activity of PC12 cells, indicating that CART peptide does not participate in differentiation or neuronal activity. CART peptide increased the phosphorylation of SAPK/JNK (stress-activated protein kinase/c-Jun-amino-terminal kinase) and subsequent c-Jun protein expression. These effects were reversed by SP600125, a specific JNK-kinase inhibitor. CART peptide did not significantly affect ERK (extracellular signal-regulated kinase), CREB (cAMP responsive element binding protein), or p38 phosphorylation and c-Fos protein expression. Central administration of CART peptide into mice also resulted in increased c-Jun positive cells in dorsomedial hypothalamic nucleus and nucleus of the solitary tract, areas involved in food intake regulation. Activation of c-Jun by CART peptide might indicate a possible role of CART peptide in managing stress conditions rather than a role in cell proliferation or differentiation as well as the more complex and/or specific regulation ways by transcription factors in some nuclei involved in food intake regulation. The characteristics of stress that CART peptide potentially mediates should be further studied.

• MeSH
• acetylcholinesterasa analýza   MeSH
• buňky PC12 MeSH
• hypothalamus účinky léků   metabolismus   MeSH
• krysa rodu rattus MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• nucleus solitarius účinky léků   metabolismus   MeSH
• proteiny nervové tkáně metabolismus   farmakologie   MeSH
• receptory peptidů metabolismus   MeSH
• signální transdukce fyziologie   MeSH
• vazebná místa MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In this study we analyzed molecular mechanisms of antioxidative protection of thymol and thyme oil using differentiated PC12 cells, a widely accepted neuronal model. Thymol due to multiple functions is commonly used for clinical applications. However, its action on nervous tissue remains poorly understood. We evaluated the effect of 24 h incubation of the cells with thymol (100 and 400 μM) and equivalent content of thyme oil. Microsomal glutathione transferase 1 (Mgst1) is an important player in anti-oxidative protection because of its transferase and peroxidase activities. Since its expression decreases during aging, we also used stable transfected cells with downregulated Mgst1 (PC12_M). We analyzed cell viability, lipid peroxidation level, glutathione content and expression of key enzymes responsible for cellular reduced glutathione – catalytic subunit of γ-glutamylcysteine ligase and glutathione reductase. Whereas thymol and thyme oil improved antioxidant capacity of control cells, diminished protection was observed in PC12_M line. Increasing interest in natural dietary components has focused attention on plants used as a rich source of bioactive phytochemicals. However, currently available information on the safe amount for thyme oil using appears to be insufficient. Our results indicate that the thyme oil should be used with caution, especially by elderly people.

• MeSH
• antioxidancia * fyziologie   chemie   MeSH
• buňky PC12 účinky léků   MeSH
• glutathion fyziologie   účinky léků   MeSH
• glutathiontransferasa MeSH
• neurony účinky léků   MeSH
• oleje rostlin * farmakologie   chemie   MeSH
• peroxidace lipidů účinky léků   MeSH
• stárnutí účinky léků   MeSH
• statistika jako téma MeSH
• techniky in vitro statistika a číselné údaje   MeSH
• thymol * farmakologie   MeSH
• Thymus (rostlina) MeSH
• viabilita buněk účinky léků   MeSH
• Publikační typ
• hodnotící studie MeSH
• práce podpořená grantem MeSH

Ischemic stroke is a severe cause of disability and death all over the world. To search for effective therapy for ischemic stroke, PC12 cells damaged by oxygenation and glucose deprivation/restoration were employed to assess the protective effects of inotodiol. As a result, inotodiol can improve the cell viability and attenuate the leakage of lactate dehydrogenase. Meanwhile, inotodiol can prevent oxidative stress by reducing reactive oxygen species generation, decreasing the content of malonic dialdehyde, and increasing the activity of superoxide dismutase. In addition, the dysfunction of mitochondria induced by oxygenation and glucose deprivation/restoration was ameliorated through decreasing the level of intracellular calcium and increasing the mitochondrial membrane potential. At the same time, inotodiol can inhibit PC12 cells apoptosis through downregulation of Caspase-3 and Bax as well as upregulation of Bcl-2. These results reveal inotodiol can protect PC12 cells against the injury induced by oxygenation and glucose deprivation/restoration. This investigation gives promising evidences for the therapy of ischemic stroke.

• Klíčová slova
• inotodiol,
• MeSH
• apoptóza MeSH
• buňky PC12 * patologie   účinky léků   MeSH
• cévní mozková příhoda * farmakoterapie   MeSH
• glukosa MeSH
• kaspasa 3 MeSH
• krysa rodu rattus MeSH
• kyslík MeSH
• laktátdehydrogenasy metabolismus   MeSH
• lanosterol analogy a deriváty   farmakologie   MeSH
• membránový potenciál mitochondrií účinky léků   MeSH
• oxidační stres účinky léků   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• vápníková signalizace účinky léků   MeSH
• western blotting MeSH
• Check Tag
• krysa rodu rattus MeSH

Reperfusion therapies for ischaemic stroke can induce secondary injury accompanied by neuronal death. The Y-box binding protein 1 (YBX1), an oncoprotein, is critical for regulating tumour cell proliferation and apoptosis. Thus, we wanted to know whether YBX1 could regulate neuronal cell apoptosis caused by cerebral ischaemia/reperfusion (I/R). We established a model of cerebral I/R-induced injury in vitro by oxygen-glucose deprivation/reoxygenation (OGD/R) treatment and determined YBX1 expression using Western blot. Next, the effect of YBX1 on the apoptosis and viability of OGD/R-treated PC12 cells was evaluated by flow cytometry, MTT assay, and Western blot. Besides, the release of lactate dehydrogenase (LDH) and the activity of catalase (CAT) and superoxide dismutase (SOD) were detected to evaluate oxidative stress of PC12 cells induced by OGD/R. The regulatory roles of YBX1 in the AKT/GSK3β pathway were examined by Western blot. As a result, OGD/R treatment down-regulated YBX1 expression in PC12 cells. YBX1 over-expression attenuated the growth inhibition and apoptosis of PC12 cells induced by OGD/R. Besides, the increase of LDH release and the decrease of SOD and CAT activities caused by OGD/R were reversed by YBX1 over-expression. Moreover, YBX1 over-expression could activate the AKT/GSK3β pathway in OGD/ R-treated PC12 cells. Therefore, YBX1 could protect against OGD/R-induced injury in PC12 cells through activating the AKT/GSK3β signalling pathway, and thus YBX1 has the potential to become a therapeutic target for cerebral I/R-induced injury.

• MeSH
• apoptóza MeSH
• buňky PC12 MeSH
• cévní mozková příhoda * MeSH
• DNA vazebné proteiny MeSH
• glukosa MeSH
• ischemie mozku * MeSH
• kinasa glykogensynthasy 3beta * MeSH
• krysa rodu rattus MeSH
• kyslík MeSH
• oxidační stres MeSH
• protoonkogenní proteiny c-akt * MeSH
• reperfuzní poškození * MeSH
• viabilita buněk MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Alzheimer's disease (AD) threatens to become the scourge of the 21st century, hence there is an urgent need for the exploration of multipotent drug in the treatment of AD. Rhizophora mucronata, tropical red mangrove has been widely used in traditional oriental medicine as astringent, antiseptic, antibacterial, anti-ulcerogenic and anti-inflammatory agent. The present study aimed to evaluate the neuroprotective effect of the catechin rich methanolic leaf extract of R. mucronata (MERM), against Aβ-induced neurotoxicity. Exposure of PC12 cells to Aβ (25–35) increased cellular oxidative stress, the number of apoptotic cells and caspase-3 activity ultimately leading to neuronal death. Pre-treatment with MERM (50 μg/ml) significantly attenuated cell death, decreased the level of intracellular reactive oxygen and nitrogen species thereby inhibiting lipid peroxidation and protein oxidation. MERM attenuated Aβ (25–35) induced apoptosis by stabilizing the mitochondrial membrane potential, and inhibiting caspase-3 activity. MERM also restored the antioxidant status of the Aβ (25–35) treated cells by effectively scavenging ROS/RNS species. HPTLC analysis of MERM illustrated the presence of (+)-catechin as major constituent. Results conclude that MERM effectively attenuated the neurotoxicity induced by Aβ-associated oxidative stress, implying that MERM can act as a potent drug for the treatment of AD.

• MeSH
• Alzheimerova nemoc farmakoterapie   MeSH
• amyloidní beta-protein * účinky léků   MeSH
• apoptóza MeSH
• buňky PC12 * účinky léků   MeSH
• chromatografie na tenké vrstvě MeSH
• katechin MeSH
• kultivované buňky MeSH
• listy rostlin MeSH
• membránový potenciál mitochondrií MeSH
• neuroprotektivní látky farmakologie   chemická syntéza   chemie   MeSH
• oxidační stres MeSH
• peptidové fragmenty účinky léků   MeSH
• reaktivní formy kyslíku MeSH
• Rhizophoraceae * MeSH
• rostlinné extrakty MeSH
• Publikační typ
• práce podpořená grantem MeSH

Labile redox-active iron ions have been implicated in various neurodegenerative disorders, including the Parkinson's disease (PD). Iron chelation has been successfully used in clinical practice to manage iron overload in diseases such as thalassemia major; however, the use of conventional iron chelators in pathological states without systemic iron overload remains at the preclinical investigative level and is complicated by the risk of adverse outcomes due to systemic iron depletion. In this study, we examined three clinically-used chelators, namely, desferrioxamine, deferiprone and deferasirox and compared them with experimental agent salicylaldehyde isonicotinoyl hydrazone (SIH) and its boronate-masked prochelator BSIH for protection of differentiated PC12 cells against the toxicity of catecholamines 6-hydroxydopamine and dopamine and their oxidation products. All the assayed chelating agents were able to significantly reduce the catecholamine toxicity in a dose-dependent manner. Whereas hydrophilic chelator desferrioxamine exerted protection only at high and clinically unachievable concentrations, deferiprone and deferasirox significantly reduced the catecholamine neurotoxicity at concentrations that are within their plasma levels following standard dosage. SIH was the most effective iron chelator to protect the cells with the lowest own toxicity of all the assayed conventional chelators. This favorable feature was even more pronounced in prochelator BSIH that does not chelate iron unless its protective group is cleaved in disease-specific oxidative stress conditions. Hence, this study demonstrated that while iron chelation may have general neuroprotective potential against catecholamine auto-oxidation and toxicity, SIH and BSIH represent promising lead molecules and warrant further studies in more complex animal models.

• MeSH
• buňky PC12 MeSH
• chelátory železa * farmakologie   MeSH
• deferasirox farmakologie   MeSH
• deferipron farmakologie   MeSH
• deferoxamin farmakologie   MeSH
• dopamin farmakologie   MeSH
• katecholaminy farmakologie   MeSH
• krysa rodu rattus MeSH
• oxidační stres MeSH
• oxidopamin farmakologie   MeSH
• přetížení železem * MeSH
• železo farmakologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

• MeSH
• acetylcholinesterasa metabolismus   MeSH
• cholin-O-acetyltransferasa metabolismus   MeSH
• krysa rodu rattus MeSH
• nádory nadledvin patologie   MeSH
• neurotrofní faktory farmakologie   MeSH
• peroxid vodíku farmakologie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH

• MeSH
• buněčná smrt fyziologie   MeSH
• buňky PC12 MeSH
• mozková hypoxie a ischemie patofyziologie   MeSH
• Publikační typ
• kongresy MeSH