Az anti-apoptosis mechanizmus vizsgálata agyi ischaemia/hypoxia modellekben = Studies on anti-apoptosis mechanisms in models of brain ischaemia/hypoxia

A hypoxia indukálta sejt nekrózis/apoptózis kivédése vagy mérséklése igéretes kutatási terület az alkalmas terápiás eljárások bevezetésére. In vitro és in vivo modellekben vizsgáltuk a (-)-deprenyl, az antiapoptotikus gént tartalmazó adenovírus vektorok és az ösztrogén hatását a hypoxia/ischemia indukálta változásokon. Neuronális PC12 sejtkultúrában és hippocampus organotipikus tenyészetében a deprenyl csökkenti a nekrózis és apoptózis mértékét, javítja a neuronális regenerációt. Indukálja az antiapoptotikus Bcl-2 és SOD-1 fehérjéket és a GAP-43 plaszticitás proteint. Védi a sejteket az oxidatív károsodástól és fenntartja a mitokondriális membránpotenciált. Vizsgálataink szerint a deprenyl neuroprotektív hatását mitokondriális hatásokon keresztül fejti ki. Az antiapoptotikus Bcl-2 vagy Bcl-XL gént tartalmazó adenovírus vektorok PC12 kultúrában védik a mitokondriális funkciót, csökkentik az apoptotikus sejtek arányát és növelik a GAP-43 expressziót. Az antiapoptotikus génbevitel a cytoprotektív hatás mellett fokozza a javító gének (synapsin-1, nestin, c-fos) expresszióját is. Az ösztrogén neuroprotektív hatása jól ismert, de a repair folyamatokra gyakorolt hatása nem. Nagy-dózisú ösztrogén adás hatását vizsgáltuk gerbilen átmeneti agyi ischemiaban. Adataink azt mutatják, hogy az ösztrogén nem csak neuroprotektív, hanem növeli a GAP-43 és a nestin plaszticitás gének expresszióját, ami kifejezésre jut az állatok jobb viselkedés vizsgálati eredményeiben. | Hypoxia induced cell necrosis and/or apoptosis can be moderated or prevented by antiapoptotic therapies. In in vitro and in vivo models, the impact of (-)-deprenyl, antiapoptotic gen transfer and oestrogen were tested on hypoxia/ischemia induced changes. In neuronal PC12 cell culture and orgatypic hippocampal slice culture, deprenyl reduces necrosis and apoptosis and induces neuronal repair via induction of antiapoptotic Bcl-2, SOD-1 proteins and GAP-43 plasticity protein. Deprenyl protects cells from oxidative damage and helps to mantain mitochondrial membrane potencial. Our new findings support the assumption that the neuroprotective effect of deprenyl is related to the mitochondrial processes. In PC12 culture, the antiapoptotic Bcl-2 and Bcl-XL genes transferred to adenovirus vector protect the mitochondrial fuction, reduce the number of apoptotic cells and augment the expression of GAP-43 protein. The antiapoptotic gene delivery is not only cytoprotective moreover augments the expression of repair genes synapsin-1, nestin, c-fos after hypoxic insults. Neuroprotective effect of oestrogen is well documented however its effect on repair mechanisms is still not elucidated. We investigated the effect of the high-dose oestrogen therapy on cerebral plasticity after transient forebrain ischemia in gerbils. Our novel finding is that oestrogen is not just neuroprotective but increases the expression of plasticity genes GAP-43 and nestin which results better performance of gerbils in behaviour tests

Eating habits modulate short term memory and epigenetical regulation of brain derived neurotrophic factor in hippocampus of low- and high running capacity rats

Exercise capacity and dietary restriction (DR) are linked to improved quality of life, including enhanced brain function and neuro-protection. Brain derived neurotrophic factor (BDNF) is one of the key proteins involved in the beneficial effects of exercise on brain. Low capacity runner (LCR) and high capacity runner (HCR) rats were subjected to DR in order to investigate the regulation of BDNF. HCR-DR rats out-performed other groups in a passive avoidance test. BDNF content increased significantly in the hippocampus of HCR-DR groups compared to control groups (p<0.05). The acetylation of H3 increased significantly only in the LCR-DR group. However, chip-assay revealed that the specific binding between acetylated histone H3 and BNDF promoter was increased in both LCR-DR and HCR-DR groups. In spite of these increases in binding, at the transcriptional level only, the LCR-DR group showed an increase in BDNF mRNA content. Additionally, DR also induced the activity of cAMP response element-binding protein (CREB), while the content of SIRT1 was not altered. Peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) was elevated in HCR-DR groups. But, based on the levels of nuclear respiratory factor-1 and cytocrome c oxidase, it appears that DR did not cause mitochondrial biogenesis. The data suggest that DR-mediated induction of BDNF levels includes chromatin remodeling. Moreover, DR does not induce mitochondrial biogenesis in the hippocampus of LCR/HCR rats. DR results in different responses to a passive avoidance test, and BDNF regulation in LCR and HCR rats

SIRT1 may play a crucial role in overload-induced hypertrophy of skeletal muscle

Silent mating type information regulation 2 homologue 1 (SIRT1) activity and content increased significantly in overload-induced hypertrophy. SIRT1-mediated signalling through Akt, the endothelial nitric oxide synthase mediated pathway, regulates anabolic process in the hypertrophy of skeletal muscle. The regulation of catabolic signalling via forkhead box O 1 and protein ubiquitination is SIRT1 dependent. Overload-induced changes in microRNA levels regulate SIRT1 and insulin-like growth factor 1 signalling. Significant skeletal muscle mass guarantees functional wellbeing and is important for high level performance in many sports. Although the molecular mechanism for skeletal muscle hypertrophy has been well studied, it still is not completely understood. In the present study, we used a functional overload model to induce plantaris muscle hypertrophy by surgically removing the soleus and gastrocnemius muscles in rats. Two weeks of muscle ablation resulted in a 40% increase in muscle mass, which was associated with a significant increase in silent mating type information regulation 2 homologue 1 (SIRT1) content and activity (P < 0.001). SIRT1-regulated Akt, endothelial nitric oxide synthase and GLUT4 levels were also induced in hypertrophied muscles, and SIRT1 levels correlated with muscle mass, paired box protein 7 (Pax7), proliferating cell nuclear antigen (PCNA) and nicotinamide phosphoribosyltransferase (Nampt) levels. Alternatively, decreased forkhead box O 1 (FOXO1) and increased K48 polyubiquitination also suggest that SIRT1 could be involved in the catabolic process of hypertrophy. Furthermore, increased levels of K63 and muscle RING finger 2 (MuRF2) protein could also be important enhancers of muscle mass. We report here that the levels of miR1 and miR133a decrease in hypertrophy and negatively correlate with muscle mass, SIRT1 and Nampt levels. Our results reveal a strong correlation between SIRT1 levels and activity, SIRT1-regulated pathways and overload-induced hypertrophy. These findings, along with the well-known regulatory roles that SIRT1 plays in modulating both anabolic and catabolic pathways, allow us to propose the hypothesis that SIRT1 may actually play a crucial causal role in overload-induced hypertrophy of skeletal muscle. This hypothesis will now require rigorous direct and functional testing.National Strength and Conditioning Association OTKA. Grant Number: 112810 Hungarian Academy of Science National Institute of Environmental Health Sciences. Grant Number: ES00359

DNA methylation clock DNAmFitAge shows regular exercise is associated with slower aging and systemic adaptation

DNAmPhenoAge, DNAmGrimAge, and the newly developed DNAmFitAge are DNA methylation (DNAm)-based biomarkers that reflect the individual aging process. Here, we examine the relationship between physical fitness and DNAm-based biomarkers in adults aged 33–88 with a wide range of physical fitness (including athletes with long-term training history). Higher levels of VO 2 max ( ρ = 0.2, p = 6.4E − 4, r = 0.19, p = 1.2E − 3), Jumpmax ( p = 0.11, p = 5.5E − 2, r = 0.13, p = 2.8E − 2), Gripmax ( ρ = 0.17, p = 3.5E − 3, r = 0.16, p = 5.6E − 3), and HDL levels ( ρ = 0.18, p = 1.95E − 3, r = 0.19, p = 1.1E − 3) are associated with better verbal short-term memory. In addition, verbal short-term memory is associated with decelerated aging assessed with the new DNAm biomarker FitAgeAcceleration ( ρ : − 0.18, p = 0.0017). DNAmFitAge can distinguish high-fitness individuals from low/medium-fitness individuals better than existing DNAm biomarkers and estimates a younger biological age in the high-fit males and females (1.5 and 2.0 years younger, respectively). Our research shows that regular physical exercise contributes to observable physiological and methylation differences which are beneficial to the aging process. DNAmFitAge has now emerged as a new biological marker of quality of life

Reverse regulation of endothelial cells and myointimal hyperplasia on cell proliferation by a heatshock protein-coinducer after hypoxia.

BACKGROUND AND PURPOSE: Myointimal hyperplasia (MIH) cells are related to permanent upregulated proliferation as tumor-like cells. The aim of this study is to assess whether treatment of cells after hypoxia by Iroxanadine heat-shock protein (HSP-coinducer) predicts recovery through cell proliferation. METHODS: Vascular smooth muscle cells (VSMC) and brain capillary endothelial cells (HBEC) were isolated from human origin and MIH-cells from early carotid restenosis after surgery. Cell proliferation was quantified by bromuridine (BrdU) incorporation after hypoxia/reoxygenation. HSP72 and cyclin-dependent kinase (CDKN1A) mRNA expression was assessed by reverse transcription-polymerase chain reaction (PCR) and cell cycle distribution by flow cytometry (FACS) analysis. RESULTS: After hypoxia/reoxygenation, the proliferation of MIH-cells increased, whereas endothelial cells decreased (MIH: 0.266+/-0.016 versus 0.336+/-0.024; P<0.05; HBEC: 1.249+/-0.10 versus 0.878+/-0.11; P<0.05). Whereas augmented proliferation of MIH-cells was reduced (40% to 45%) by HSP-coinducer, diminished HBEC proliferation increased (46.2%). Stress-activated-protein-kinase (SAPK)p38-dependent cell cycle redistribution was generated by an increase in HSP72 and CDKN1A mRNA levels in MIH-cells. CONCLUSIONS: The 2 key players of early restenosis (MIH, EC) were oppositely regulated and correspondingly after treatment by HSP-coinducer reverse recovered. Drug candidate may have therapeutic potential in (re)restenosis

SIRT1 may play a crucial role in overload-induced hypertrophy of skeletal muscle

Significant skeletal muscle mass guarantees functional wellbeing and is important for high level performance in many sports. Although the molecular mechanism for skeletal muscle hypertrophy has been well-studied, it still is not completely understood. In the present study, we used a functional overload model to induce plantaris muscle hypertrophy by surgically removing the soleus, and gastrocnemius muscles in rats. Two weeks of muscle ablation resulted in a 40% increase in muscle mass, which was associated with a significant increase in SIRT1 content and activity (P < 0.001). SIRT1-regulated Akt, eNOS, GLUT4 levels were also induced in hypertrophied muscles, and SIRT1 levels correlated with muscle mass, paired box protein 7 (Pax7), proliferating cell nuclear antigen (PCNA) and nicotinamide phosphoribosyltransferase (Nampt) levels. Alternatively, decreased FOXO1 and increased K48 polyubiquitination also suggest that SIRT1 could also be involved in the catabolic process of hypertrophy. Furthermore, increased levels of K63 and muscle RING finger 2 (MuRF2) protein could also be important enhancers of muscle mass. We report here that the levels of miR1 and miR133a decrease in hypertrophy and negatively correlate with muscle mass, SIRT1, and Nampt levels. Our results reveal a strong agreement between SIRT1 levels and activity, SIRT1 regulated pathways, and overload-induced hypertrophy. These findings, along with the well-known regulatory roles that SIRT1 plays in modulating both anabolic and catabolic pathways, allow us to propose the hypothesis that SIRT1 may actually play a crucial causal role in overload induced hypertrophy of skeletal muscle. This hypothesis will now require rigorous direct and functional testing. This article is protected by copyright. All rights reserved

Exercise combined with postbiotics treatment results in synergistic improvement of mitochondrial function in the brain of male transgenic mice for Alzheimer’s disease

Abstract Background It has been suggested that exercise training and postbiotic supplement could decelerate the progress of functional and biochemical deterioration in double transgenic mice overexpresses mutated forms of the genes for human amyloid precursor protein (APPsw) and presenilin 1 (m146L) (APP/PS1TG). Our earlier published data indicated that the mice performed better than controls on the Morris Maze Test parallel with decreased occurrence of amyloid-β plaques in the hippocampus. We investigated the neuroprotective and therapeutic effects of high-intensity training and postbiotic supplementation. Methods Thirty-two adult APP/PS1TG mice were randomly divided into four groups: (1) control, (2) high-intensity training (3) postbiotic, (4) combined (training and postbiotic) treatment for 20 weeks. In this study, the whole hemibrain without hippocampus was used to find molecular traits explaining improved brain function. We applied qualitative RT-PCR for gene expression, Western blot for protein level, and Zymography for LONP1 activity. Disaggregation analysis of Aβ-40 was performed in the presence of Lactobacillus acidophilus and Bifidobacterium longum lysate. Results We found that exercise training decreased Alzheimer’s Disease (AD)-related gene expression (NF-kB) that was not affected by postbiotic treatment. The preparation used for postbiotic treatment is composed of tyndallized Bifidobacterium longum and Lactobacillus acidophilus. Both of the postbiotics effectively disaggregated amyloid-β/Aβ-40 aggregates by chelating Zn2+ and Cu2+ ions. The postbiotic treatment decreased endogenous human APPTG protein expression and mouse APP gene expression in the hemibrains. In addition, the postbiotic treatment elevated mitochondrial LONP1 activity as well. Conclusion Our findings revealed distinct mechanisms behind improved memory performance in the whole brain: while exercise training modulates NF-kB signaling pathway regulating immune response until postbiotic diminishes APP gene expression, disaggregates pre-existing amyloid-β plaques and activates mitochondrial protein quality control in the region of brain out of hippocampus. Using the above treatments complements and efficiently slows down the development of AD

Novel Possible Pharmaceutical Research Tools: Stem Cells, Gene Delivery and their Combination.

Both stem cell research and gene delivery are very promising fields of today's biomedical research. In the present review we first attempt to summarize the state of the art in stem cell research. We describe the major categories of stem cells based on cell sources: embryonic, fetal, postnatal and induced pluripotent stem cells. We then present new data on stem cell cultures of dental pulp origin as examples of the progress of postnatal stem cell research. Afterwards, we briefly summarize the most promising achievements in the field of gene delivery. As an example of such advances, we describe novel in vitro and in vivo gene delivery studies to demonstrate that salivary glands are highly potential targets for gene therapy: they can be used to produce therapeutic peptides delivered either into the oral cavity or into the systemic circulation. Finally, we describe and compare studies combining the use of stem cells and gene delivery. We conclude that stem cell therapy and gene delivery alone are both very exciting research areas, and they may act in synergy when used in combination

DNA methylation clock DNAmFitAge shows regular exercise is associated with slower aging and systemic adaptation

DNAmPhenoAge, DNAmGrimAge, and the newly developed DNAmFitAge are DNA methylation (DNAm)-based biomarkers that reflect the individual aging process. Here, we examine the relationship between physical fitness and DNAm-based biomarkers in adults aged 33–88 with a wide range of physical fitness (including athletes with long-term training history). Higher levels of VO 2 max ( ρ = 0.2, p = 6.4E − 4, r = 0.19, p = 1.2E − 3), Jumpmax ( p = 0.11, p = 5.5E − 2, r = 0.13, p = 2.8E − 2), Gripmax ( ρ = 0.17, p = 3.5E − 3, r = 0.16, p = 5.6E − 3), and HDL levels ( ρ = 0.18, p = 1.95E − 3, r = 0.19, p = 1.1E − 3) are associated with better verbal short-term memory. In addition, verbal short-term memory is associated with decelerated aging assessed with the new DNAm biomarker FitAgeAcceleration ( ρ : − 0.18, p = 0.0017). DNAmFitAge can distinguish high-fitness individuals from low/medium-fitness individuals better than existing DNAm biomarkers and estimates a younger biological age in the high-fit males and females (1.5 and 2.0 years younger, respectively). Our research shows that regular physical exercise contributes to observable physiological and methylation differences which are beneficial to the aging process. DNAmFitAge has now emerged as a new biological marker of quality of life