Background. Neurotransmitter adenosine and B-group vitamins have neuroprotective, remyelinizing and anti-neuroinflammatory properties. Despite the studies of these molecules for decades, the molecular mechanisms of their synergistic effect on neuroinflammation processes are unexplored and not systematized.Objective: to establish the molecular mechanisms of synergism of adenosine, thiamine, niacin and cyanocobalamin in counteracting the pathology of diabetic polyneuropathy (DPN).Material and methods. The molecular mechanisms of action of adenosine, thiamine (vitamin B1), niacin (vitamin PP) and cyanocobalamin (vitamin B12) in the pathophysiology of DPN were determined using functional analysis of genomic and proteomic databases.Results. The analysis of 20,180 annotated proteins of the human proteome identified 504 vitamin-PP-dependent, 22 vitamin-B1-dependent, 24 vitamin-B12-dependent and 50 adenosine-dependent proteins. The proteins of the human proteome were detected, the activity or levels of which are important for reducing neuroinflammation, remyelination, neurogenesis, biosynthesis of neuronal adenosine triphosphate, myelin homeostasis, neuroplasticity, neutralization of homocysteine, regeneration of nerve fibers and maintaining the endothelium of the microvascular bed.Conclusion. The discovered molecular mechanisms of synergism of the studied molecules are of fundamental importance for comprehension of the processes of neuroinflammation regulation and remyelination to prevent diabetic polyneuropathy and other neurodegenerative diseases.

Alzheimer's-like disease was simulated in female adult Wistar CRL(WI) WUBR rats by 6-week intragastric administration of aluminum chloride at a dose of 200 mg/kg body mass. In the presence of the developed oxidative stress (OS), we found a decrease in the activities of the tricarboxylic acid cycle (TCA cycle) enzymes and an increase in the activities of the pentose phosphate pathway (PPP) dehydrogenases, as well as a reduction of SH-and SS-groups in proteins along with an increased SH/SS ratio and glutathionylation with simultaneous decreases of glutathione (GSH) and the GSH/GSSG ratio and its redox potential. The glutathione system enzymes were changed multidirectionally, with glutathione reductase remaining stable. Decreased activities of GSH biosynthesis enzymes and cysteine content were noticed. The intragastric administration of the CoA biosynthesis modulators D-panthenol (PL), D-pantethine, or D-homopantothenate (HPA) at a dose of 200 mg/kg from the fifth week of the experiment caused either reduction or leveling of OS manifestations in blood plasma, an increase in acetyl cholinesterase, normalization of the activities of TCA cycle and PPP enzymes and the P-SH level (not the SH/SS ratio), and a significant reduction of S-glutathionylation, as well as increases in the GSH level, the GSH/GSSG ratio and the redox potential in the hemispheres. The effect of CoA system modulators was manifested in activation of glutathione transferase, a decrease of glutathione peroxidase and less evident activation of GSH biosynthesis enzymes (that is, PL) although they contributed to the elevation of the cysteine content due to reduced protein S-cysteinylation. The levels and the ratio of CoA/acetyl-CoA (except for PL) were not changed by toxicosis and the OS modulators. The feasibility of nonconenzyme effects was confirmed by the administration of HPA. The phenomenon of redox activity of the CoA biosynthesis modulators with clearly directional effects on the glutathione system and the TCA cycle and PPP enzymes during alleviation of OS and aluminum neurotoxicosis is discussed.

Alzheimer’s-like disease was simulated in female adult Wistar CRL(WI) WUBR rats by 6-week intragastric administration of aluminium chloride at a dose of 200 mg/kg body mass. In the presence of developed oxidative stress (OS), we found a decrease in the activities of tricarboxylic acid cycle (TCA cycle) enzymes and an increase in the activities of pentose phosphate pathway (PPP) dehydrogenases as well as a reduction of SH-and SS-groups in proteins (P) along with the increased SH/SS ratio and glutathionylation with simultaneous decreases of glutathione (GSH) and the GSH/GSSG ratio and its redox potential in the brain hemispheres. The glutathione system enzymes were changed multidirectionally, with glutathione reductase remaining stable. Decreased activities of GSH biosynthesis enzymes and cysteine content were noticed. The intragastric administration of the CoA biosynthesis modulators D-panthenol (PL), D-pantethine or D-homopantothenate (HPA) at a dose of 200 mg/kg since the 5th week of the experiment caused either reduction or leveling of OS manifestations in blood plasma, an increase in acetyl cholinesterase, normalization of the activities of TCA cycle and PPP enzymes, P-SH level (not the SH/SS ratio) and a considerable reduction of S-glutathionylation as well as increases in GSH level, the GSH/GSSG ratio and redox potential in the hemispheres. The effect of CoA system modulators was manifested in activation of glutathione transferase, a decrease of glutathione peroxidase and less evident activation of GSH biosynthesis enzymes (PL) although they contributed to the elevation of cysteine content due to the reduced protein S-cysteinylation. The levels and the ratio of CoA/acetyl-CoA (except for PL) were not changed by toxicosis and the OS modulators. The feasibility of non-conenzyme effects was confirmed by the administration of HPA. The phenomenon of redox activity of the CoA biosynthesis modulators with clearly directional effects on the glutathione system and the TCA cycle and PPP enzymes during alleviation of OS and aluminium neurotoxicosis is discussed.

The changes in the parameters of oxidative stress, energy metabolism, and redox potential of the glutathione system in the rat brain following cerebral ischemia were studied. To correct metabolic disorders, the pantothenic acid derivatives were used in combination with precursors of glutathione biosynthesis and selenium substances.Cerebral ischemia was modeled by ligating the both common carotid arteries in rats for 2 h. Drugs were administered i.p. in the following doses: panthenol – 400 mg/kg, N-acetylcysteine – 150, nanoselen – 1 mg/kg, three times: 1 h before ligation of the carotid arteries, at the time of ligation and 1 hour after ligation. We showed that the development of oxidative stress caused by ischemia is accompanied by the changes in the parameters of energy metabolism and the pentose phosphate pathway in the cerebral hemispheres. Simultaneously, there are a decrease in the GSH level, an increase in the GSSG content, a decrease in the GSH/GSSG ratio, and the activation of enzymes of redox transformations of glutathione.The redox potential of the glutathione system decreases and shifts towards oxidation, while the level of S-glutathionylated proteins increases. Thus, the value of the GSH/GSSG ratio and the protein glutathionylation intensity are the sensitive indicators of the redox potential in the brain tissue and can be used as markers of the extent of changes in the redox balance. The panthenol injection to animals leads to a decrease in the content of free radical oxidation products, violations of oxidative phosphorylation and restoration of thiol-disulfide balance in the brain. When panthenol is administered together with N-acetylcysteine and nanoselen, the corrective effect of panthenol is enhanced.

The possibility of using vitamin D to normalize the histological structure of the spleen changed under the influence of prednisolone in rats was studied. The animals were subjected to the intragastric administration of saline and either prednisolone (5 mg/kg b. w.) or its combination with vitamin D (800 IU/kg) daily for 3 weeks. The results of morphometric analysis of spleen slices reveal that the administration of prednisolone leads to a significant decrease in spleen sizes, white pulp volume, and in sizes of germinal centers of lymphatic follicles within the white pulp, and to an increase in the number of megakaryocytes in the red pulp. Vitamin D alleviates histological changes due to the prednisolone treatment, in particular the substantial restoration in sizes of germinal centers in the spleen has been found. The data obtained suggest the benefits of further studies of possible mechanisms of vitamin D to normalize immunological and hematopoietic functions in subjects subjected to glucocorticoid treatment.

Free-radical-mediated processes are involved in a variety of physiological events, while oxidative stress and related redox deregulation are implicated in various pathological events. Tripeptide glutathione plays an important role in the antioxidant defense of the brain, particularly in the maintenance of the optimal redox state in neurons and glial cells. We studied the combined effects of pantothenic acid derivatives, pantothenol and calcium pantothenate, and memantine, which is a glutamate receptor antagonist that is widely used for the treatment of dementia, on amnesia induced by scopolamine in rats. Scopolamine induced amnesia in rats; however, unexpectedly, this effect was even more expressed in the memantine-pretreated animals. Memory impairments were less manifested in the rats that were pretreated with memantine in combination with panthenol or calcium pantothenate. The detrimental scopolamine effect on memory was accompanied by significant depletions of glutathione and coenzyme A in the brain. While memantine recovered the glutathione status to some extent, it nevertheless further aggravated the scopolamine influence on coenzyme A levels. An alleviation of scopolamine-induced memory impairments that was observed after combined pretreatment with memantine and panthenol or calcium pantothenate was accompanied by a normalization of coenzyme A levels, while the effects on glutathione redox did not correlate with the behavioral data.

Indicators of oxidative stress (OS), systemic inflammation, metabolism and redox status of glutathione (GSH) were investigated and compared in patients with ST-segment elevation myocardial infarction on electrocardiograms (STEMI), and patients with unstable angina (UA). The elevated and decreased myeloperoxidase level, superoxide dismutase activity, and moderate increased plasma levels of interleukin-6, while maintaining the antioxidant potential, were found in Group 1. Disorders in pro-/antioxidant balance and systemic inflammatory response were manifested in UA. Increased GSH concentration (and total GSH) in erythrocytes has been established for STEMI patients and the decreased GSH for UA patients. Thus, a significant shift of erythrocytes redox to oxidization and increase (unlike STEMI patients) of glutathione peroxidase activity were recorded. Mechanisms of the pro- and antioxidant functions of red blood cells in acute coronary syndrome are considered. The role of red blood cell glutathione to provide more oxidized intravascular environment for S-glutathionylation and optimization of redox signaling in target cells is pronounced.

In this study, the effects of interoceptive stress on the indices of free-radical oxidation and the corticosterone contents in the brain and blood of male rats were examined. In addition, we studied the effects of the GABA-like nootropic drug calcium hopanthenate and panthenol on these indices after stress. Interoceptive stress was induced by intraperitoneal injection of bacterial lipopolysaccharide and the studies were performed 24 h after injection. The derivatives of pantothenic acid substantially attenuated corticosterone accumulation in the hippocampus after interoceptive stress induced by systemic immune challenge but did not prevent a stress-induced increase in the level of substances that reacted with thiobarbituric acid in the blood of the animals.

Изучали влияние дипептида “глицил-пролин” (3 мг/кг) на изменения содержания нейроактивных аминокислот (аспартата, глицина, аланина, таурина, ГАМК, глутамата), активности ферментов ГАМК-шунта (ГАМК-трансаминазы, дегидрогеназы янтарного полуальдегида, глутаматдекарбоксилазы) и метаболизма глутамата (глутаматдегидрогеназы, аланин- и аспартат- трансаминаз) в больших полушариях мозга крыс после 6 ч ишемии мозга при трех способах введения препарата: внутрибрюшинном, субконьюктивальном и интраназальном. Установлено, что гли-про проявляет наиболее выраженное корригирующее действие в отношении ишемических нарушений при интраназальном способе введения.

Objective. To study and compare indicators of systemic inlammation and red blood cells glutathione redox potential by chronic oxidative stress (OS) in patients with coronary heart disease (CHD), type 2 diabetes mellitus (T2DM) and the combination of both diseases.Design and methods. The study included 35 patients with CHD and newly diagnosed T2DM, 109 patients with CHD, 19 patients with T2DM, and 89 healthy individuals. Systemic inlammation indicators were measured including concentration of interleukin (IL) 6 and 8, myeloperoxidase (MPO) and OS in plasma, oxidized (GSSG) and reduced (GSH) glutathione, as well as their ratio and redox potential (E), the activity of glutathione reductase (GR) and glutathione peroxidase (GP) in red blood cells.Results. Signiicant growth of systemic inlammation (IL-6, IL-8, MPO) indicators on the weakening antioxidant defense enzyme has been registered in patients with a combination of CHD and T2DM, a 3,4-fold reduction of GSH level, 5,8-fold of 2GSH/GSSH in red blood cells and E values towards the oxidized state at 36,3 mV were noted. These changes were less pronounced in CHD group and were absent in T2DM group. All subjects showed a decreased activity of GP and increased level of glycosylated hemoglobin. The correlation of E and activity GR have been found in patients with carbohydrate metabolism disorder. High positive relationship between IL concentration and red blood cells GR, E indicators has been identiied in patients with CHD and T2DM. There is a need to control and correct cellular redox potential in patients with CHD and T2DM.Conclusion. The combination of CHD and T2DM contributes to the decrease of red blood cells redox status glutathione secondary to the increased systemic inlammatory response and chronic OS indicators.

В работе проведено исследование эффектов интероцептивного стресса на показатели свободнорадикального окисления и уровень кортикостерона в мозге и крови самцов крыс. Кроме того, изучали влияние ГАМК-подобного ноотропа гопантената кальция и пантенола на эти показатели при стрессе. Интероцептивный стресс индуцировали интраперитонеальной инъекцией бактериального липополисахарида; исследование проводили через 24 ч после инъекции. Производные пантотеновой кислоты существенно снижали селективное накопление кортикостерона в гиппокампе при интероцептивном стрессе, вызванном системной иммунной провокацией, но не предотвращали вызванное стрессом накопление продуктов, реагирующих с тиобарбитуровой кислотой, в крови животных.

We studied the effects of the glycyl-proline dipeptide (Gly-Pro) at a dose of 3 mg/kg on the contents of neuroactive amino acids, including aspartate, glycine, alanine, taurine, GABA, and glutamate, the activity of the enzymes of the GABA shunt, such as GABA-transaminase, succinic semialdehyde dehydrogenase, and glutamate decarboxylase, and enzymes of glutamate metabolism, such as glutamate dehydrogenase and alanine and aspartate transaminases in the neocortex of rats subjected to 6-h brain ischemia. We used three means of dipeptide administration, including intraperitoneal, subconjunctival, and intranasal. We found that the protective anti-ischemic effect of Gly-Pro was mostly expressed after intranasal administration.

Проведено исследование морфологических изменений и экспрессии маркеров астроглии и микроглии в сенсомоторной коре и гиппокампе крыс, перенесших хроническое невротизирующее воздействие. Установлено, что адаптация к хроническому стрессу сопровождается возникновением большого числа поврежденных нейронов как в слое V сенсомоторной коры, так в пирамидном слое гиппокампа, причем наиболее выражены эти изменения в поле СА3. Неврозоподобное состояние у крыс характеризуется увеличением экспрессии маркеров микроглии в гиппокампе, которое сохраняется в течение месяца после окончания стрессорного воздействия. Экспрессия астроцитарного маркера GFAP снижается в поле СА3 гиппокампа. Введение производного пантотеновой кислоты – пантенола, способствует стабилизации глиального ответа на хронический стресс, но его эффекты не длительны и не могут предотвратить активацию микроглии в постстрессорный период.

We studied the morphological changes and expression of the astroglial and microglial markers in the sensorimotor cortex and hippocampus of rats that were subjected to chronic stress. Chronic neurotization was associated with the appearance of a large number of damaged neurons in layer V of the sensorimotor cortex and the pyramidal layer of the hippocampus. These changes were considerably expressed in the CA3 field. A neurosis-like state in rats was accompanied by an increased expression of microglial markers in the hippocampus and this effect was evident even 1 month after the end of stress. The expression of the astroglial marker GFAP decreased in the CA3 hippocampal field. Treatment of animals with the pantothenic acid derivative panthenol stabilized the glial response to chronic stress; however, its effects were not long-lasting and did not prevent activation of microglia during the period after stress.

In vivo study of neuroprotective effects of D-panthenol has been performed on a rat model of middle cerebral artery occlusion (MCAO). Administration of panthenol (i.p., 200 mg/kg 24 h, 0.5 h before and 0.5 h, 24 h, 48 h after MCAO) significantly ameliorated neurological deficit 1–5 days after MCAO, reduced body weight loss and cortical damage induced by 30 min MCAO. The results are the first in vivo evidence for neuroprotective properties of D-panthenol and suggest that it has a good potential for treatment of neurological diseases.

We injected albino rats intraperitoneally with D-[ 3 H]-pantethine at a dose of 15 mg/kg of body weight (0.014 Ci/mmol) or perfused rat hippocampal slices with 47 μM (1.3 Ci/mmol) solution of this substance. Using reversed phase HPLC, we found that metabolic products of D-pantethine, including phosphopantothenate and pantothenate, were rapidly absorbed from blood serum and retained in the hippocampus. We also observed their limited biotransformation in the system of coenzyme A (CoA) biosynthesis with the formation of 4′-phosphopantetheine and CoA 10 min to 24 h after radionuclide administration. Hippocampal slices were perfused for 3–7 min. In the fractions that were eluted with 8.5% methanol, we found 4′-phosphopantetheine/pantetheine (64–47%), pantothenate (18–15%), phosphopantothenate (9–18%), and CoA (7–10%). The decrease in the radionuclide-retaining capacity in the slices was associated with a linear increase in the 4′-phosphopantothenate fraction. The role of 4′-phosphopantothenate in the stabilization of the CoA level and mechanisms of neuroprotection in the hippocampus, as well as the potential for the use of pantethine-based drugs in neurological practice are discussed.

Calcium homopantothenate (CHP, pantogam, or hopantenic acid) at a dose of 100 μg/kg was injected into white female Wistar rats weighing 180–210 g for 4 weeks. Aluminum chloride (two times at a dose of 190 mg/kg) and D-pantenol (seven times at a dose of 200 mg/kg) were injected to a subgroup of rats during the last 2 weeks of the experiment. CHP did not change the level of coenzyme A (CoA) in the hippocampus, brainstem, and cerebellum, while aluminum neurotoxicity resulted in a decrease in the CoA-SH level in the hippocampus and acyl-CoA level in the cerebellum. A decrease in glutathione level, redox status, and the total amount of glutathione and glutathione peroxidase activity was observed during aluminum neurotoxicity. Both treatments activated glutathione reductase and decreased the activity of glucose-6-phosphate dehydrogenase in the brainstem. These changes were eliminated by pantenol. The latter increased the redox status of CNS proteins in the case of the combination of treatment with CHP and aluminum neurotoxicity.

Parameters of the redox status of the synaptosomal glutathione system and the functional state of the synaptosomal membrane of rat brain were investigated after activation of free radical processes in vitro. A decrease in the content of reduced glutathione along with a simultaneous increase in that of its oxidized form and inhibition of Na+,K+-ATPase and acetylcholine esterase were observed for the model of oxidative stress. These parameters correlated with changes in the total phospholipids: cholesterol ratio and in the balance of thiol- and disulphide groups of proteins of synaptosomal membranes. Preincubation of synaptosomes with 0.5 mM sodium pantothenate prevented the negative effects of the Fenton reaction, but had no effect in the presence of the buthionine-sulfoximine inhibitor of glutathione biosynthesis. Our results provide evidence for the regulatory effect of the glutathione redox status on the structural and functional state of synaptosomal membranes, and for its possible modulation by the CoA biosynthetic precursor.