Brain ischemia is a leading cause of death and disability worldwide that occurs when blood supply of the brain is disrupted. Brain-derived neurotrophic factor (BDNF) is a protective factor in neurodegenerative conditions. Nevertheless, there are some problems when exogenous BDNF is to be used in the clinic. 14-3-3ζ is a pro-survival highly-expressed protein in the brain that protects neurons against death. This study evaluates 14-3-3ζ effects on BDNF transcription at early time point after ischemia and its possible protective effects against ischemia damage. Human 14-3-3ζ protein was purified after expression. Rats were assigned into four groups, including sham, ischemia, and two treatment groups. Stereotaxic cannula implantation was carried out in the right cerebral ventricle. After one week, rats underwent middle cerebral artery occlusion (MCAO) surgery and received 14-3-3ζ (produced in our laboratory or standard form as control) in the middle of ischemia time. At 6 h of reperfusion after ischemia, brain parts containing the hippocampus, the cortex, the piriform cortex-amygdala and the striatum were collected for real time PCR analysis. At 24 h of reperfusion after ischemia, neurological function evaluation and infarction volume measurement were performed. The present study showed that 14-3-3ζ could up-regulate BDNF mRNA at early time point after ischemia in the hippocampus, in the cortex and in the piriform cortex-amygdala and could also improve neurological outcome and reduce infarct volume. It seems that 14-3-3ζ could be a candidate factor for increasing endogenous BDNF in the brain and a potential therapeutic factor against brain ischemia.

• MeSH
• dospělí MeSH
• infarkt arteria cerebri media farmakoterapie   metabolismus   patologie   MeSH
• ischemie mozku * metabolismus   MeSH
• izoformy RNA metabolismus   účinky léků   MeSH
• laboratorní zvířata MeSH
• modely nemocí na zvířatech MeSH
• neuroprotektivní látky MeSH
• potkani Wistar MeSH
• proteiny 14-3-3 farmakologie   terapeutické užití   MeSH
• receptor trkB metabolismus   účinky léků   MeSH
• vazba proteinů MeSH
• Check Tag
• dospělí MeSH
• mužské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakty MeSH

Cíl studie: Zhodnotit neuroprotektivní účinek dantrolenu a jeho mechanizmus. Materiál a metodologie: Byl vytvořen model okluze střední mozkové tepny (middle cerebral artery occlusion, MCAO) a pomocí dvojitého fluorescenčního barvení zhodnocena exprese proteinu vážícího 12-kDa FK506 (FKBP12) a ryanodinového receptoru (RyR). Objem nekrotické tkáně byl zhodnocen pomocí barvení trifenyltetrazolium chloridem (TTC) a toluidinovou modří. Výsledky: Studie prokázala upregulaci FKBP12 ve skupině s dantrolenem ve srovnání s kontrolní skupinou ve všech časových bodech; RyR se exprimoval společně s FKBP12, přestože se mezi hodinou 1 a 24 jeho zbarvení zvýšilo jen mírně. Rovněž jsme zjistili, že léčba dantrolenem (1 hod a 4 hod po MCAO) nesnížila objem infarktové tkáně (p > 0,05); léčba dantrolenem (24 hod po MCAO) snížila objemy nekrotické tkáně a měla významný vliv na objem infarktové tkáně (p < 0,05); což naznačuje, že inhibice ryanodinového receptoru (RyR) vede k cytoprotekci. Závěr: FKBP12 může hrát důležitou roli v procesu neuronálního přežití a smrti po mozkové ischemii, a neuroprotektivní účinky dantrolenu mohou být dány upregulací FKBP12 a snížením objemu infarktové tkáně.

Purpose: To investigate any neuroprotection provided by dantrolene and the mechanism involved in its action. Materials and methods: A middle cerebral artery occlusion (MCAO) model was followed and the expressions of 12-kDa FK506-binding protein (FKBP12) and ryanodine receptor were assessed by fluorescent double staining. Infarct volume was assessed by triphenyltetrazolium chloride (TTC) and toluidine blue staining. Results: The study showed that FKBP12 was upregulated in the dantrolene group compared with the control group at each corresponding time point; RyR co-expressed with FKBP12, although its staining only increased slightly from 1 h to 24 h. We also found that treatment with dantrolene 1 h and 4 h after MCAO did not reduce infarct volumes (p >0.05); however, treatment with dantrolene 24 h after MCAO reduced infarct volumes and had a significant effect on the infarct volume (p <0.05), indicating that inhibition of ryanodine receptor (RyR) leads to cytoprotection. Conclusion: FKBP12 may play an important role in the processes of neuronal survival or death following cerebral ischemia, and dantrolene may exert its neuroprotective effects by upregulating FKBP12 and thereby decreasing infarct volume.

• Klíčová slova
• FKBP12, ryanodinový receptor, mozková ischemie,
• MeSH
• barvení a značení MeSH
• dantrolen analogy a deriváty   terapeutické užití   MeSH
• experimenty na zvířatech MeSH
• financování organizované MeSH
• fluorescenční protilátková technika MeSH
• ischemie mozku farmakoterapie   MeSH
• malformace mozkové kůry diagnóza   MeSH
• mozková kůra zranění   MeSH
• neuroprotektivní látky klasifikace   terapeutické užití   MeSH
• potkani Wistar MeSH
• proteiny vázající takrolimus farmakologie   terapeutické užití   MeSH
• reperfuze metody   škodlivé účinky   využití   MeSH
• ryanodinový receptor vápníkového kanálu farmakologie   terapeutické užití   MeSH
• statistika jako téma MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH

• MeSH
• astrocyty fyziologie   MeSH
• cytokiny fyziologie   genetika   MeSH
• ischemie mozku patofyziologie   MeSH
• krysa rodu rattus MeSH
• modely u zvířat MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• kongresy MeSH

100 rats were randomly divided into a sham-operated group and middle cerebral artery occlusion (MCAO) modeling groups. The sham group after surgery was observed for 14 days. After MCAO, some rats received isometric contraction training (ICT) which was as follows: an atraumatic tourniquet was placed around left or right hind limb to achieve hind limb ischemia for 5 min, followed by 5 min of reperfusion, 4 cycles for one time, once a day, and five days per week. The MCAO modeling groups included the following four groups: i) a group only received MCAO, and was observed for seven days (MCAO-7d), ii) a group only received MCAO, and was observed for 14 days (MCAO-14d), iii) a group, after MCAO, received ICT for seven days (ICT-7d), and iv) a group, after MCAO, received ICT for 14 days (ICT-14d). Brain infarct area, behavioral outcomes, the number of neurons, apoptosis, cerebral edema and cerebral water content were assessed, respectively. The mRNA expression of vascular endothelial growth factor (VEGF) was assayed with RT-PCR, and protein expression of VEGF was quantified with western blot. compared with MCAO controls, cerebral infarction, neurological deficits and neuronal apoptosis were reduced significantly in the ICT groups, while the number of neurons was increased. Moreover, the mRNA expression of VEGF and protein expression of VEGF were enhanced after 1 and 2 weeks of ICT. ICT may promote angiogenesis and neuroprotection after ischemic stroke and this new remodeling method provide a novel strategy for rehabilitation of stroke patients.

• MeSH
• cévní mozková příhoda * terapie   MeSH
• infarkt arteria cerebri media MeSH
• ischemie mozku * metabolismus   MeSH
• isometrická kontrakce * MeSH
• kondiční příprava zvířat * MeSH
• krysa rodu rattus MeSH
• messenger RNA MeSH
• modely nemocí na zvířatech MeSH
• neuroprotekce MeSH
• potkani Sprague-Dawley MeSH
• vaskulární endoteliální růstový faktor A metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Zkoumali jsme účinky dekompresní kraniektomie u krysílio modelu „maligního" infarktu střední mozkové tepny (ACM). Celkem 55 samců krys Sprague-Dawiey o váze 290 až 340 g bylo rozděleno do těchto skupin: 1) simulovane operovaná skupina (skupina 1, n = 14, jako kontrolní); 2) skupina s okluzí ACM (OACM) (skupina 2, n = 26, zůstala neléčená po permanentní OACM); 3) skupina s dekompresní kraniektomií (skupina 3, n = 15, vhodná dekomprese do 1 hodiny po OACM). Po 48 hodinách bylo zjištěno, že dekompresní kraniektomie může znamenat jednoznačný prínos z hlediska přežití, lepších neurologických výsledků, jakož i menšího rozsahu infarktu po OACM u krys. Výrazně snižuje odumírání neuronů v kůře, ne však v hlubokých struktúrách, jako je hippocampus a striatum.

We investigated the effects of decompressive craniectomy in a rat model of „malignant“ middle cerebral artery (MCA) infarction. A total of 55 male Sprague-Dawley rats weighing 290–340 g were allocated to the following groups: (1) Sham operated group (group 1, n = 14, as controls); (2) MCA occlusion (MCAO) group (group 2, n = 26, remained untreated after right permanently MCAO); (3) Decompressive craniectomy group (group 3, n = 15, received right decompression at 1 hour after MCAO). 48 hours later we found that decompressive craniectomy can offer a clear survival benefit, a better neurological score as well as a reduced infarct volume after MCAO in rats. It significantly decreases neuronal death in cortex but not in deep structures such as hippocampus and striatum.

• MeSH
• analýza přežití MeSH
• chirurgická dekomprese metody   využití   MeSH
• experimenty na zvířatech statistika a číselné údaje   MeSH
• infarkty mozkového kmene chirurgie   terapie   MeSH
• interpretace statistických dat MeSH
• kraniotomie metody   využití   MeSH
• krysa rodu rattus chirurgie   MeSH
• pooperační komplikace prevence a kontrola   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus chirurgie   MeSH
• zvířata MeSH

Akútne mozgovocievne ochorenia väčšinou klasifikované ako mozgová príhoda sú najčastejšie poruchy centrálneho nervového systému. Prechodná ložisková mozgová ischemia (stroke) vedie k rozsiahlemu excitotoxickému poškodeniu neurónov a glie. Počas ischémie je spos uvoľnením glutamátu zvýšená aj hladina mimobunkového ATP, ktoré aktivuje purínergné receptory, špeciálne ionotropné receptory P2X. Nadmerná aktivácia receptora podtypu P2X7 môže viesť k excitotoxickej bunkovej smrti. V práci sme za účelom štúdia neuroprotektívneho efektu Brilliant Blue B (BBG), špecifického inhibítora receptora P2X7, použili model prechodnej ložiskovej ischémie u potkanov navodením oklúzie strednej mozgovej tepny (MCAO). Aplikácia BBG v tomto modeli spôsobuje redukciu objemu infarktovej zóny až o 50 % v porovnaní so skupinou zvierat, ktorým bol podaný fyziologický roztok. Tieto výsledky ukazujú, že receptor P2X7 je než sprostredkovateľom poškodelia tkaniva po mozgovej príhode. To poukazuje na skupina purínergných receptorov môžu byť vhodným molekulárnym cieľom ri vývoji terapeutických postupov zmierňujúcich poškodenenie tkaniva po mozgovej príhode.

Acute cerebrovascular diseases mostly classified as ischem- ic strokes are the most frequent disorders of the central nervous system. Transient focal cerebral ischemia leads to extensive release excitotoxic neuronal and glial damage. During ischemia together with glutamate releas is increased level of extracellular ATP which activate purinergic receptors, especially ionotropic P2X receptors. Overactivation of subtype P2X7 receptors can induce excitotoxic cell death. We used middle cerebral artery occlusion (MCAO) in rats as a model of transient focal cerebral ischemia to study the neuroprotective effect of Brilliant Blue G (BBG), selective inhibitor of P2X7 receptor. Treatment with BBG produced about 50% reduction in the extent of brain damage compared to treatment with vehicle alone. These results show that P2X7 purinergic receptors mediate tissue damage in neurons following transient brain ischemia. Therefore, these receptors are a relevant molecular target for the development of new treatments to attenuate brain damage following stroke.

• Publikační typ
• abstrakt z konference MeSH

Polydendrocytes (also known as NG2 glial cells) constitute a fourth major glial cell type in the adult mammalian central nervous system (CNS) that is distinct from other cell types. Although much evidence suggests that these cells are multipotent in vitro, their differentiation potential in vivo under physiological or pathophysiological conditions is still controversial.To follow the fate of polydendrocytes after CNS pathology, permanent middle cerebral artery occlusion (MCAo), a commonly used model of focal cerebral ischemia, was carried out on adult NG2creBAC:ZEG double transgenic mice, in which enhanced green fluorescent protein (EGFP) is expressed in polydendrocytes and their progeny. The phenotype of the EGFP(+) cells was analyzed using immunohistochemistry and the patch-clamp technique 3, 7 and 14 days after MCAo. In sham-operated mice (control), EGFP(+) cells in the cortex expressed protein markers and displayed electrophysiological properties of polydendrocytes and oligodendrocytes. We did not detect any co-labeling of EGFP with neuronal, microglial or astroglial markers in this region, thus proving polydendrocyte unipotent differentiation potential under physiological conditions. Three days after MCAo the number of EGFP(+) cells in the gliotic tissue dramatically increased when compared to control animals, and these cells displayed properties of proliferating cells. However, in later phases after MCAo a large subpopulation of EGFP(+) cells expressed protein markers and electrophysiological properties of astrocytes that contribute to the formation of glial scar. Importantly, some EGFP(+) cells displayed membrane properties typical for neural precursor cells, and moreover these cells expressed doublecortin (DCX)--a marker of newly-derived neuronal cells. Taken together, our data indicate that polydendrocytes in the dorsal cortex display multipotent differentiation potential after focal ischemia.

• MeSH
• buněčná diferenciace MeSH
• diferenciační antigeny biosyntéza   MeSH
• ischemie mozku metabolismus   patologie   MeSH
• modely nemocí na zvířatech MeSH
• myši transgenní MeSH
• myši MeSH
• neuroglie metabolismus   patologie   MeSH
• proliferace buněk MeSH
• proteiny nervové tkáně biosyntéza   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The mouse Dach1 gene, involved in the development of the neocortex and the hippocampus, is expressed by neural stem cells (NSCs) during early neurogenesis, and its expression also continues in a subpopulation of cells in the dorsal part of the lateral ventricles (LV) of the adult mouse brain. In this study we aimed to elucidate the role of Dach1-expressing cells in adult neurogenesis/gliogenesis under physiological as well as post-ischemic conditions, employing transgenic mice in which the expression of green fluorescent protein (GFP) is controlled by the D6 promotor of the mouse Dach1 gene. A neurosphere-forming assay of GFP⁺ cells isolated from the dorsal part of the LV was carried out with subsequent differentiation in vitro. To elucidate the neurogenic/gliogenic potential of GFP⁺ cells in the dorsal part of the LV, in situ immunohistochemical/electrophysiological analyses of GFP⁺ cells in adult sham-operated brains (controls) and those after middle cerebral artery occlusion (MCAo) were performed. The GFP⁺ cells isolated from the dorsal part of the LV of controls formed neurospheres and differentiated solely into a glial phenotype, while those isolated after MCAo also gave rise to cells with the properties of neuronal precursors. In situ analyses revealed that GFP⁺ cells express the phenotype of adult NSCs or neuroblasts in controls as well as following ischemia. Following MCAo we found a significantly increased number of GFP⁺ cells expressing doublecortin as well as a number of GFP⁺ cells migrating through the rostral migratory stream into the olfactory bulb, where they probably differentiated into calretinin⁺ interneurons. Collectively, our results suggest the involvement of the mouse Dach1 gene in adult neurogenesis; cells expressing this gene exhibit the properties of adult NSCs or neuroblasts and respond to MCAo by enhanced neurogenesis.

• MeSH
• 4-aminopyridin farmakologie   MeSH
• blokátory sodíkových kanálů farmakologie   MeSH
• buněčná diferenciace fyziologie   MeSH
• degenerace nervu etiologie   patologie   MeSH
• dospělé kmenové buňky fyziologie   MeSH
• infarkt arteria cerebri media komplikace   MeSH
• membránové potenciály účinky léků   MeSH
• metoda terčíkového zámku MeSH
• modely nemocí na zvířatech MeSH
• myši inbrední C57BL MeSH
• myši transgenní MeSH
• myši MeSH
• neurogeneze fyziologie   MeSH
• neurony metabolismus   MeSH
• oční proteiny metabolismus   MeSH
• počet buněk MeSH
• proteiny nervové tkáně metabolismus   MeSH
• techniky in vitro MeSH
• tetraethylamonium farmakologie   MeSH
• tetrodotoxin farmakologie   MeSH
• ventriculi laterales patologie   MeSH
• zelené fluorescenční proteiny genetika   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH