Spectroscopic studies of Delayed Luminescence emitted by an in vitro model for studying of the effects of amyloid-βeta (Aβ) have been performed. Aβ is a neurotoxic protein overexpressed in Alzheimer's Disease (AD), which is also related to mitochondrial dysfunction. The experiments have been carried out on primary Olfactory Ensheathing Cells (OECs) cultures. Cells have been exposed to Aβ(1-42) native full-length peptide or to Aβ(25-35), a toxic fragment of Aβ, or Aβ(35-25), a non toxic Aβ fragment, both in absence and in presence of Astaxanthin, a well-known antioxidant agent. To monitor cell viability, MTT test was used. Reactive oxygen species and reduced glutathione levels were utilized to test the oxidative intracellular status. We also assessed the expression of some glial markers (Glial Acidic Fibrillary Protein, Vimentin), of Nestin, stem cell marker, and the activation of the apoptotic pathway assessing caspase-3 cleavage. We found that, in OECs, Glial Acidic Fibrillary Protein, Vimentin expression and caspase-3 exhibited a significant enhancement in Aβ(1–42) and Aβ(25–35) exposed cells. The pre-treatment with Astaxanthin restored the levels of Vimentin and caspase-3 to control values, increasing also Nestin expression levels and reestablished the intracellular oxidative status modified by the exposure to Aβ(1–42) or Aβ(25–35) of OECs. DL intensity and kinetics changes as a function of the treatments were also measured. In particular, an increase in DL emission, with respect the untreated cells (controls), was observed in cells exposed to Aβ(25-35) fragment. This emission appeared quenched in presence of Astaxanthin.
In the framework of the research project ETHICS “Pre-clinical experimental and theoretical studies to improve treatment and protection by charged particles” funded by the National Nuclear Physics Institute, Italy, we studied the phenomenon called delayed luminescence emitted by non-tumorigenic breast epithelial MCF10A cell line after proton irradiation at different doses (0.5, 2, 6, 9 Gy). The aim is to found possible correlations between delayed luminescence and in vitro damaging induced by ion irradiation. The first results of this research show that the delayed luminescence kinetics is proton dose dependent. An interesting correlation between delayed luminescence and clonogenic potential was observed.
The first results concerning the possibility to use Delayed Luminescence spectroscopy to evaluate the in vitro induction of cytotoxic effects on human glioblastoma cells of nanostructured lipid carrier and drug-loaded nanostructured lipid carrier are showed in this contribution. We tested the effects of nanostructured lipid carrier, ferulic acid and ferulic acidloaded nanostructured lipid carrier on U-87MG cell line. The study seems to confirm the ability of Delayed Luminescence to be sensible indicator of alterations induced on functionality of the mitochondrial respiratory chain complex I in U-87MG cancer cells when treated with nanostructured lipid carriers.
Correlation between apoptosis and UVA-induced ultraweak photon emission delayed luminescence (DL) from tumor thyroid cell lines was investigated. In particular, the effects of berberine, an alkaloid that has been reported to have anticancer activities, on two cancer cell lines were studied. The FTC-133 and 8305C cell lines, as representative of follicular and anaplastic thyroid human cancer, respectively, were chosen. The results show that berberine is able to arrest cell cycle and activate apoptotic pathway as shown in both cell lines by deoxyribonucleic acid fragmentation, caspase-3 cleavage, p53 and p27 protein overexpression. In parallel, changes in DL spectral components after berberine treatment support the hypothesis that DL from human cells originates mainly from mitochondria, since berberine acts especially at the mitochondrial level. The decrease of DL blue component for both cell lines could be related to the decrease of intra-mitochondrial nicotinamide adenine dinucleotide and may be a hallmark of induced apoptosis. In contrast, the response in the red spectral range is different for the two cell lines and may be ascribed to a different iron homeostasis.
Irina Baran, Diana Ionescue, Simona Privitera, Agata Scordino, Maria Mocanu, Francesco Musumeci, Rosaria Grasso, Marisa Gulino, Adrian Iftime, Ioana Tofolean, Alexandru Garaiman, Alexandru Goicea, Ruxandra Irimia, Alexandru Dimancea, Constanta Ganea
The role of mitochondrial complex I in ultraweak photon-induced delayed photon emission [delayed luminescence (DL)] of human leukemia Jurkat T cells was probed by using complex I targeting agents like rotenone, menadione, and quercetin. Rotenone, a complex I-specific inhibitor, dose-dependently increased the mitochondrial level of reduced nicotinamide adenine dinucleotide (NADH), decreased clonogenic survival, and induced apoptosis. A strong correlation was found between the mitochondrial levels of NADH and oxidized flavin mononucleotide (FMN ox ) in rotenone-, menadione- and quercetin-treated cells. Rotenone enhanced DL dose-dependently, whereas quercetin and menadione inhibited DL as well as NADH or FMN ox . Collectively, the data suggest that DL of Jurkat cells originates mainly from mitochondrial complex I, which functions predominantly as a dimer and less frequently as a tetramer. In individual monomers, both pairs of pyridine nucleotide (NADH/reduced nicotinamide adenine dinucleotide phosphate) sites and flavin (FMN-a/FMN-b) sites appear to bind cooperatively their specific ligands. Enhancement of delayed red-light emission by rotenone suggests that the mean time for one-electron reduction of ubiquinone or FMN-a by the terminal Fe/S center (N2) is 20 or 284 μs, respectively. All these findings suggest that DL spectroscopy could be used as a reliable, sensitive, and robust technique to probe electron flow within complex I in situ.
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