Glutathione oxidized (L-Glutathione oxidized)

Measurement of oxidized/reduced glutathione ratio

Glutathione (GSH) is the most abundant antioxidant in aerobic cells, present in micromolar (microM)-concentrations in bodily fluids and in millimolar (mM) concentrations in tissue. GSH is critical for protecting the brain from oxidative stress, acting as a free radical scavenger and inhibitor of lipid peroxidation. GSH also participates in the detoxification of hydrogen peroxide by various glutathione peroxidases. The ratio of reduced GSH to oxidized GSH (GSSG) is an indicator of cellular health, with reduced GSH constituting up to 98% of cellular GSH under normal conditions. However, the GSH/GSSG ratio is reduced in neurodegenerative diseases, such as Parkinson's disease (PD) and Alzheimer's disease (AD). Measuring the GSH/GSSG ratio in pathological tissues and experimental models thereof in comparison to the results in controls is an excellent way to assess potential therapeutics efficacy in maintaining cellular redox potential. The availability of UV/Visible instruments equipped with 96-well plate readers as common laboratory equipment has made measuring the GSH/GSSG ratio on multiple samples a manageable procedure.

The role of glutathione redox imbalance in autism spectrum disorder: A review

The role of glutathione in autism spectrum disorder (ASD) is emerging as a major topic, due to its role in the maintenance of the intracellular redox balance. Several studies have implicated glutathione redox imbalance as a leading factor in ASD, and both ASD and many other neurodevelopmental disorders involve low levels of reduced glutathione (GSH), high levels of oxidized glutathione (GSSG), and abnormalities in the expressions of glutathione-related enzymes in the blood or brain. Glutathione metabolism, through its impact on redox environment or redox-independent mechanisms, interferes with multiple mechanisms involved in ASD pathogenesis. Glutathione-mediated regulation of glutamate receptors [e.g., N-methyl-d-aspartate (NMDA) receptor], as well as the role of glutamate as a substrate for glutathione synthesis, may be involved in the regulation of glutamate excitotoxicity. However, the interaction between glutathione and glutamate in the pathogenesis of brain diseases may vary from synergism to antagonism. Modulation of glutathione is also associated with regulation of redox-sensitive transcription factors nuclear factor kappa B (NF-κB) and activator protein 1 (AP-1) and downstream signaling (proinflammatory cytokines and inducible enzymes), thus providing a significant impact on neuroinflammation. Mitochondrial dysfunction, as well as neuronal apoptosis, may also provide a significant link between glutathione metabolism and ASD. Furthermore, it has been recently highlighted that glutathione can affect and modulate DNA methylation and epigenetics. Review analysis including research studies meeting the required criteria for analysis showed statistically significant differences between the plasma GSH and GSSG levels as well as GSH:GSSG ratio in autistic patients compared with healthy individuals (P = 0.0145, P = 0.0150 and P = 0.0202, respectively). Therefore, the existing data provide a strong background on the role of the glutathione system in ASD pathogenesis. Future research is necessary to investigate the role of glutathione redox signaling in ASD, which could potentially also lead to promising therapeutics.

Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method

The spectrophotometric/microplate reader assay method for glutathione (GSH) involves oxidation of GSH by the sulfhydryl reagent 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB) to form the yellow derivative 5'-thio-2-nitrobenzoic acid (TNB), measurable at 412 nm. The glutathione disulfide (GSSG) formed can be recycled to GSH by glutathione reductase in the presence of NADPH. The assay is composed of two parts: the preparation of cell cytosolic/tissue extracts and the detection of total glutathione (GSH and GSSG). The method is simple, convenient, sensitive and accurate. The lowest detection for GSH and GSSG is 0.103 nM in a 96-well plate. This method is rapid and the whole procedure takes no longer than 15 min including reagent preparation. The method can assay GSH in whole blood, plasma, serum, lung lavage fluid, cerebrospinal fluid, urine, tissues and cell extracts and can be extended for drug discovery/pharmacology and toxicology protocols to study the effects of drugs and toxic compounds on glutathione metabolism.

Efficient Esterification of Oxidized l-Glutathione and Other Small Peptides

Oxidized l-glutathione was esterified to the tetra methyl ester using thionyl chloride in methanol solvent. Other alcohols were tested and the reaction progress was monitored via ESI-MS. This procedure proved to be compatible with other small peptides not containing serine and cysteine residues. In contrast to previously reported methods this procedure provided convenient access to esterified peptides requiring no purification, extended reaction times, or complicated reaction setups.

A Yellow Fluorescence Probe for the Detection of Oxidized Glutathione and Biological Imaging

It is well-known that the ratio of reduced l-glutathione (GSH) to oxidized l-glutathione (GSSG) is a vital biomarker for monitoring overall cellular health, thus detecting the intracellular concentration of glutathione is of great significance. Recently, an increasing number of reports have published various methods for GSH detection, but studies on the detection of GSSG are still rare. Here, we report a kind of new yellow fluorescent carbon dots (CDs) for the detection of GSSG through a fluorescence "off-on" process. Because the surface is rich in amino groups, the CDs show a positive potential. When the concentration of GSSG was continuously increased, the CDs' fluorescence dropped sharply, while the fluorescence gradually recovered after the addition of sodium sulfide. The phenomenon of fluorescence quenching is linear with the concentration of the quencher (GSSG)(0-200 μM), and 0.18 μM is calculated as the detection limit. More interestingly, as a fluorescent probe, the CDs can be further used for fluorescence imaging in living cells and zebrafish.