The extracellular redox state modulates mitochondrial function, gluconeogenesis, and glycogen synthesis in murine hepatocytes.

Circulating redox state changes, determined by the ratio of reduced/oxidized pairs of different metabolites, have been associated with metabolic diseases. However, the pathogenic contribution of these changes and whether they modulate normal tissue function is unclear. As alterations in hepatic gluconeogenesis and glycogen metabolism are hallmarks that characterize insulin resistance and type 2 diabetes, we tested whether imposed changes in the extracellular redox state could modulate these processes. Thus, primary hepatocytes were treated with different ratios of the following physiological extracellular redox couples: ß-hydroxybutyrate (ßOHB)/acetoacetate (Acoc), reduced glutathione (GSH)/oxidized glutathione (GSSG), and cysteine/cystine. Exposure to a more oxidized ratio via extracellular ßOHB/Acoc, GSH/GSSG, and cysteine/cystine in hepatocytes from fed mice increased intracellular hydrogen peroxide without causing oxidative damage. On the other hand, addition of more reduced ratios of extracellular ßOHB/Acoc led to increased NAD(P)H and maximal mitochondrial respiratory capacity in hepatocytes. Greater ßOHB/Acoc ratios were also associated with decreased ß-oxidation, as expected with enhanced lipogenesis. In hepatocytes from fasted mice, a more extracellular reduced state of ßOHB/Acoc led to increased alanine-stimulated gluconeogenesis and enhanced glycogen synthesis capacity from added glucose. Thus, we demonstrated for the first time that the extracellular redox state regulates the major metabolic functions of the liver and involves changes in intracellular NADH, hydrogen peroxide, and mitochondrial respiration. Because redox state in the blood can be communicated to all metabolically sensitive tissues, this work confirms the hypothesis that circulating redox state may be an important regulator of whole body metabolism and contribute to alterations associated with metabolic diseases.

[Experimental study of the therapeutic efficiency of the lithium salt of glutathione disulphide in the conditions of acute external gamma-irradiation].

Experimental studies on white outbred male mice subjected to gamma-irradiation at doses LD(80/30) have found that the therapeutic use of Litan contributes to the increase (by 40-50%) in the survival rate of the animals. Introduction to irradiated animals of Litan increases the intensity of the inclusion of 3H-thymidine in the DNA of the bone marrow cells and is associated with a growing number of splenic endogenous colonies, which can testify to the stimulating influence of the drug on the proliferation and differentiation of hematopoietic cells, as well as on the deficit restoration of full-fledged mature cell forms in the peripheral blood.

Phase 2 study of neoadjuvant treatment with NOV-002 in combination with doxorubicin and cyclophosphamide followed by docetaxel in patients with HER-2 negative clinical stage II-IIIc breast cancer.

NOV-002 (a formulation of disodium glutathione disulfide) modulates signaling pathways involved in tumor cell proliferation and metastasis and enhances anti-tumor immune responsiveness in tumor models. The addition of NOV-002 to chemotherapy has been shown to increase anti-tumor efficacy in animal models and some early phase oncology trials. We evaluated the clinical effects of NOV-002 in primary breast cancer, whether adding NOV-002 to standard preoperative chemotherapy increased pathologic complete response rates (pCR) at surgery, and determined whether NOV-002 mitigated hematologic toxicities of chemotherapy and whether levels of myeloid derived suppressor cells (MDSC) were predictive of response. Forty-one women with newly diagnosed stages II-IIIc HER-2 negative breast cancer received doxorubicin-cyclophosphamide followed by docetaxel (AC â†’ T) every 3 weeks and concurrent daily NOV-002 injections. The trial was powered to detect a doubling of pCR rate from 16 to 32% with NOV-002 plus AC â†’ T (α = 0.05, ß = 80%). Weekly complete blood counts were obtained as well as circulating MDSC levels on day 1 of each cycle were quantified. Of 39 patients with 40 evaluable tumors, 15 achieved a pCR (38%), meeting the primary endpoint of the trial. Concurrent NOV-002 resulted in pCR rates for AC â†’ T chemotherapy higher than previously reported. Patients with lower levels of circulating MDSCs at baseline and on the last cycle of chemotherapy had significantly higher probability of a pCR (P = 0.02). Further evaluation of NOV-002 in a randomized study is warranted.

Intracerebroventricular injection of glutathione and its derivative induces sedative and hypnotic effects under an acute stress in neonatal chicks.

Glutathione-related enzymes glyoxalase 1 and glutathione reductase 1 regulates anxiety in mice. To clarify the central function of glutathione as a neurotransmitter in the stress reaction, the effect of intracerebroventricular (i.c.v.) injection of reduced (GSH) (0.5, 1, 2 micromol) and oxidized (GSSG) glutathione (0.25, 0.5, 1 micromol) were investigated under an isolation-induced stress in the neonatal chick. Both GSH and GSSG dose-dependently decreased distress vocalizations and induced sleep-like behavior in chicks under acute stressful conditions. However, which glutathione is actually responsible for inducing sleep was unclear since glutathione cycles between GSH and GSSG in which two tripeptides are linked by a disulfide bond. Therefore, the behavior of chicks was monitored following the i.c.v. injection of S-methylglutathione (SMG) (0.0625, 0.25, 1 micromol). SMG does not form a disulfide bond due to the methylation of the SH group of the cysteine moiety. SMG had similar effects as observed in GSH and GSSG. In conclusion, glutathione and its derivative have sedative and hypnotic effects, and might be effective in improving psychic stress such as anxiety.

The disposition and bioavailability of 35S-GSH from 35S-GSSG in BSS PLUS in rabbit ocular tissues.

The purpose of this study was to evaluate the biodistribution and uptake of 35S-GSH into intraocular tissues following the administration of BSS PLUS containing 35S-GSSG by either an anterior chamber or intravitreal injection. This study evaluated the disposition and uptake of the 35S-radiolabel, the intracellular concentrations of 35S-GSH from extracellular 35S-GSSG, and the percentage of 35S-GSH to the total cellular GSH pool. Glutathione was analyzed by high-performance liquid chromatography (HPLC) using fluorescence detection after derivitizing the thiols in situ with monobromobimane. The effluent from the GSH peak was then collected for measurement of 35S-GSH. After an anterior chamber injection of 35S-BSS PLUS, 35S-radioactivity rapidly disappeared from the aqueous humor between 0.5 and 2 hours; corneal 35S-radioactivity remained constant over time. 35S-GSH was detected in the iris and ciliary body. However, in the cornea, 35S-GSH became the predominant radioactive thiol in the stroma, endothelium, and epithelium; the corneal stroma appeared to be a possible GSH reservoir for the adjacent corneal layers. After an intravitreal injection, 35S-radioactivity slowly decreased in the vitreous humor but was readily taken up by the tissues of the posterior segment, especially the retina and choroid, which showed the greatest concentrations of 35S-GSH of all tissues studied. The data from this study demonstrate that 35S-GSSG in BSS PLUS is metabolized and taken up by ocular cells and that 35S-GSH becomes incorporated into the intracellular GSH pool of ocular tissues.

Oxidized glutathione promotes sleep in rabbits.

Glutatione is implicated in sleep regulation. There are circadian changes in brain glutathione levels, and nocturnal intracerebroventricular (i.c.v.) slow infusion of oxidized glutathione (GSSG) or reduced glutathione (GSH) promotes rapid-eye-movement sleep (REMS) and non-REMS (NREMS) in rats. In the present experiments, we tested the effects of GSSG on duration of sleep, NREMS intensity, and brain temperature in another species, rabbits. Male New Zealand rabbits were injected with isotonic NaCl on a baseline day and one dose of GSSG on the test day [0.15, 1.5, 15, and 150 microg/rabbit, i.c.v., or 1.5 or 15 mg/kg intravenously (i.v.)]. Electroencephalogram (EEG), motor activity, and brain temperature were recorded for 6 h. Injection of 15 microg GSSG i.c.v. significantly increased the time spent in NREMS in the first 3 h after the injection. Injection of 0.15, 1.5, and 150 microg i.s.v. GSSG, as well as systemic injections of GSSG did not affect NREMS. Intensity of NREMS as measured by EEG slow-wave activity during NREMS, and brain temperature were not affected by any of the treatments. These results are consistent with the hypothesis that glutathione may be a sleep-inducing factor in the brain.