- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
Burden of obesity is increasing in the contemporary world. Although multifactorial in origin, appropriate mitochondrial function of adipocytes emerges as a factor essential for healthy adipocyte differentiation and adipose tissue function. Our study aimed to evaluate mitochondrial functions of human adipose-derived mesenchymal stem cells committed to adipogenesis. On days 0, 4, 10, and 21 of adipogenesis, we have characterized adipocyte proliferation and viability, quantified lipid accumulation in maturing cells, performed qualitative and quantitative analysis of mitochondria, determined mitochondrial respiration of cells using high-resolution respirometry, and evaluated mitochondrial membrane potential. In the course of adipogenesis, mitochondrial oxygen consumption progressively increased in states ROUTINE and E (capacity of the electron transfer system). State LEAK remained constant during first days of adipogenesis and then increased probably reflecting uncoupling ability of maturing adipocytes. Citrate synthase activity and volume of mitochondrial networks increased during differentiation, particularly between days 10 and 21. In addition, lipid accumulation remained low until day 10 and then significantly increased. In conclusion, during first days of adipogenesis, increased mitochondrial respiration is needed for transition of differentiating cells from glycolytic to oxidative metabolism and clonal expansion of preadipocytes and then more energy is needed to acquire typical metabolic phenotype of mature adipocyte.
- MeSH
- adipogeneze * MeSH
- buněčné dýchání MeSH
- kultivované buňky MeSH
- lidé MeSH
- membránový potenciál mitochondrií MeSH
- metabolismus lipidů MeSH
- mezenchymální kmenové buňky fyziologie MeSH
- mitochondrie metabolismus MeSH
- tukové buňky metabolismus MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Studie se zaměřuje na hodnocení účinku opakované expozice HBO na mitochondriální respiraci lidských fetálních plicních fibroblastů (HFL1). Mitochondriální spotřeba kyslíku byla hodnocena metodou high-resolution respirometry (HRR), životaschopnost buněk hodnocena PrestoBlue®, morfologie analyzovaná rutinní fluorescenční mikroskopií a fázovým kontrastem a byla zhodnocena aktivita superoxid dismutázy (SOD) a citrát syntázy (CS). Buňky byly vystaveny HBO (3ATA) 2 hodiny denně po dobu 5 po sobě jdoucích dní. Den po poslední HBO expozici vykazovaly buňky podstatně menší plochu a obvod, sníženou životaschopnost a zvýšenou aktivitu SOD. Nebyly zjištěny žádné změny v aktivitě CS ani v kvalitě mitochondriální sítě. HRR odhalila sníženou mitochondriální spotřebu kyslíku, která se projevila snížením aktivity komplexu II a sníženou spotřebou kyslíku při oxidací mastných kyselin. Naše zjištění dokládají, že v podmínkách napodobujících intermitentní expozici HBO, trpí plicní fibroblasty zhoršenou mitochondriální respirací spojenou s komplexem II a poruchou buněčného růstu i při zvýšené antioxidační obraně. Mechanismus této HBO-indukované mitochondriální dysfunkce je nutné hlouběji prozkoumat.
Study aims to evaluate effects of repeated exposure to HBO on mitochondrial respiration assessed by high-resolution respirometry (HRR), cell viability estimated by PrestoBlue® reaction, morphology analyzed by routine phase contrast and fluorescent microscopy, and superoxide dismutase (SOD) and citrate synthase (CS) activities using human lung fibroblasts. The cells were exposed to HBO (3 ATA) for 2 hours per day for 5 consecutive days. One day after the last exposure, HBO cells displayed significantly smaller area and perimeter, compromised viability and elevated SOD activity. No changes were detected in CS activity or quality of mitochondrial network. HRR revealed impaired mitochondrial oxygen consumption manifested by increased leak respiration, decreased activity of complex II and compromised ATP-related oxygen consumption when fatty acids were oxidized. Our findings document that in conditions mimicking chronic intermittent exposure to HBO, lung fibroblasts suffer from compromised mitochondrial respiration linked to complex II and impaired cellular growth in spite of increased antioxidant defense. Underlying mechanism of this HBO-induced mitochondrial dysfunction should be further explored.
- Klíčová slova
- respirometrie,
- MeSH
- buněčné dýchání MeSH
- fibroblasty * enzymologie metabolismus MeSH
- fluorescenční mikroskopie MeSH
- hyperbarická oxygenace * škodlivé účinky MeSH
- hyperoxie MeSH
- lidé MeSH
- mitochondrie MeSH
- plíce anatomie a histologie MeSH
- plod MeSH
- superoxiddismutasa MeSH
- techniky in vitro MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Hyperbaric oxygen (HBO) therapy, i.e. breathing pure oxygen under increased environmental pressures serves as a treatment for diverse medical conditions. However, elevated oxygen concentration can be detrimental to central nervous system or lungs. Our study aimed to evaluate the effects of repeated exposure to HBO on mitochondrial respiration assessed by high-resolution respirometry (HRR), cell viability estimated by PrestoBlue® reaction, morphology analyzed by routine phase contrast and fluorescent microscopy, and superoxide dismutase (SOD) and citrate synthase (CS) activities using human lung fibroblasts. The cells were exposed to HBO for 2 h per day for 5 consecutive days. One day after the last exposure, HBO cells displayed significantly smaller area and perimeter, compromised viability and elevated SOD activity. No changes were detected in CS activity or quality of mitochondrial network. HRR revealed impaired mitochondrial oxygen consumption manifested by increased leak respiration, decreased activity of complex II and compromised ATP-related oxygen consumption when fatty acids were oxidized. Our findings document that in conditions mimicking chronic intermittent exposure to HBO, lung fibroblasts suffer from compromised mitochondrial respiration linked to complex II and impaired cellular growth in spite of increased antioxidant defense. Underlying mechanism of this HBO-induced mitochondrial dysfunction should be further explored.
- MeSH
- buněčné dýchání fyziologie MeSH
- buněčné linie MeSH
- fibroblasty metabolismus MeSH
- hyperbarická oxygenace škodlivé účinky MeSH
- lidé MeSH
- mitochondrie fyziologie MeSH
- oxidační stres fyziologie MeSH
- plíce cytologie metabolismus MeSH
- spotřeba kyslíku fyziologie MeSH
- viabilita buněk fyziologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Evidence indicating, that persistent organic pollutants are involved in the development of obesity, has emerged. The aim of this study was to reveal whether an environmental bioaccumulative human adipose tissue contaminant, 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE), affects adipocyte differentiation. Our study was conducted on an in vitro adipogenic model of human adipose derived mesenchymal stem cells (hADMSC). The adipose cultures were exposed to DDE (concentrations: 0.1 μM, 1 μM, and 10 μM) for 28 consecutive days, from the beginning of the experiment until full differentiation. DDE was administered in lipid vehicle (NuTRIflex). Samples for gene expression analysis by RT real-time PCR were collected on days 0, 4, 10, 21 and 28 during the course of differentiation. Differentiating adipocytes cultivated in lipid-rich medium (NuTRIflex) increased the expression of perilipin 2 (PLIN2). However, the addition of DDE suppressed this effect (p < 0.03). Our results may suggest that upregulation of PLIN2, caused by exposure to lipids during the differentiation of adipocytes, is reduced in the presence of DDE. This effect of DDE warrants future attention, because of the important role of PLIN2 in formation and stabilization of lipid droplets, as the impairment of their function could be linked to the worldwide obesity epidemic.
- Klíčová slova
- perilipin-2,
- MeSH
- buněčná diferenciace účinky léků MeSH
- dichlordifenyldichlorethylen * MeSH
- kultivované buňky MeSH
- látky znečišťující životní prostředí metabolismus MeSH
- lipidová tělíska metabolismus MeSH
- membránové proteiny * genetika MeSH
- mezenchymální kmenové buňky MeSH
- neparametrická statistika MeSH
- pesticidy MeSH
- tukové buňky * metabolismus účinky léků MeSH
- upregulace MeSH
- Publikační typ
- práce podpořená grantem MeSH
- MeSH
- energetický obrat v mitochondriích fyziologie genetika MeSH
- lidé MeSH
- mitochondriální DNA genetika metabolismus MeSH
- pohybová aktivita fyziologie genetika MeSH
- reaktivní formy kyslíku metabolismus škodlivé účinky MeSH
- stárnutí buněk fyziologie MeSH
- stárnutí * fyziologie genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
Aging is a multifactorial process influenced by genetic factors, nutrition, and lifestyle. According to mitochondrial theory of aging, mitochondrial dysfunction is widely considered a major contributor to age-related processes. Mitochondria are both the main source and targets of detrimental reactions initiated in association with age-dependent deterioration of the cellular functions. Reactions leading to increased reactive oxygen species generation, mtDNA mutations, and oxidation of mitochondrial proteins result in subsequent induction of apoptotic events, impaired oxidative phosphorylation capacity, mitochondrial dynamics, biogenesis and autophagy. This review summarizes the major changes of mitochondria related to aging, with emphasis on mitochondrial DNA mutations, the role of the reactive oxygen species, and structural and functional changes of mitochondria.
- MeSH
- autofagie fyziologie MeSH
- lidé MeSH
- mitochondriální DNA fyziologie MeSH
- mitochondrie fyziologie MeSH
- oxidační stres fyziologie MeSH
- reaktivní formy kyslíku metabolismus MeSH
- stárnutí fyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
