Features of regulating delta sleep-inducing peptide by free radical processes in tissue and erythrocyte membranes in intact animals and under stress

In activity the comparative analysis of metabolic effects delta--sleep inducing peptide (DSIP) in tissues and erythrocytes of intact rats and under cold stress is conducted. The regulation effect of DSIP in attitude of free radical processes will be realised through modulation the prooxidant--antioxidant balance: both for intact animal, and at stress. Exogenous DSIP increases the antioxidant system activity in tissues of brain, liver and blood in standard conditions and under cold stress. The anti-stress effect of DSIP is directed as on increase of power endogenic enzymatic antioxidant system activity, specially glutathione peroxidase activity, and not enzymatic of antioxidant protection. The DSIP renders different influence on activity of prooxidant enzymes: for intact animal boosts the myeloperoxidase activity in blood neutrophils, not rendering essential influencing on the xanthine oxidase activity in tissues of brain, liver and activates the myeloperoxidase activity, depresses the xanthine oxidase activity for rats at stress. The membranotropic effect of DSIP in the norm and under stress is connected to increase of stability of protein--lipid interplays. The membranostabilizing effect of DSIP in conditions of stress is characterized decrease of polarity of lipid phase and negative surface charge of erythrocyte membranes, modified in course of lipid peroxidation.     

Free radical mechanism of the cold stress development in rats

Development of cold stress in rats is characterized by sharp activation of lipid peroxidation accompanied by a considerable increase of the diene conjugates level and Schiff bases in tissues of brain, liver and in erythrocytes. There is a shift in the prooxidant--antioxidant balance of the organism in the form of amplification of xanthine oxidase prooxidant enzymatic activity in the brain and liver, and a decrease of myeloperoxidase activity in blood neutrophiles of rats. The attrition at cold stress, mainly, of enzymatic endocellular antioxidant system as the result of inhibition of superoxide dismutase, catalase, glutathione reductase activities in brain, liver and erythrocytes is indemnified by activation of non-enzymatic antioxidant mechanisms. In conditions of cold stress, destabilization of erythrocyte membranes of rats described by a decrease of the microviscosity of protein-lipid contact zones and reduction of degree of immersing of proteins in lipid membrane owing to exhibiting proteins from the hydrophobic zone of membranes, or their aggregate, increase of polarity of lipid phase and negative surface charge, is marked.     

Regulation of Free Radical Processes by Delta-Sleep Inducing Peptide in Rat Tissues under Cold Stress

An intraperitoneal injection of an exogenous delta-sleep inducing peptide (DSIP) at a dose of 12 g/100 g body weight shifted the prooxidant–antioxidant balance of free radical process (FRP) in tissues and erythrocytes of rats: the activities of antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase) and the concentrations of antioxidants (reduced glutathione in particular) increased. The DSIP stimulated the myeloperoxidase activity in blood neutrophils and had no effect on the activity of xanthine oxidase, a prooxidant enzyme, in the brain and liver. Cold stress displaced the prooxidant–antioxidant balance by increasing the xanthine oxidase activity in tissues and decreasing the myeloperoxidase activity in blood neutrophils; it also inhibited the enzyme antioxidant activities in tissues and erythrocytes that was neutralized by an increased ceruloplasmin activity in blood plasma and by an elevated level of antioxidants in rat blood and tissues. Preliminary administration of DSIP to animals exposed to cold stress restored the prooxidant–antioxidant balance: it normalized the myeloperoxidase activity in blood neutrophils, decreased the xanthine oxidase activity, and increased the activity of antioxidant enzymes in tissues and erythrocytes restoring the antioxidant level. The molecular regulation mechanism of free radical processes by DSIP in tissues under stressful conditions is discussed.     

Influence of delta-sleep-inducing peptide on the activity of proteolytic enzymes in the rat brain under hypokinesia]

Single administration of the delta-sleep-inducing peptide (DSIP) in a dose of 12 micrograms/100 g to intact animals makes the activity of neutral proteinases and cathepsin D higher in the rat brain and blood serum. Hypokinesia of different duration changes activity of neutral and acidic proteinases and induces accessibility of cathepsin D to the cytosol as a result of damage in lysosomal membrane. Injection DSIP induces a decrease A/B of cathepsin D to the control level under 1-h hypokinesia condition and normalizes the neutral proteolytic activity under 6-h hypokinesia condition.     

Effects of delta-sleep-inducing peptide during ischemic injury of the rat brain

An anti-ischemic effect of the delta-sleep-inducing peptide (DSIP) was found in rats. The DSIP effect was more obvious than that of the MK-801. The data obtained is discussed considering a possible use of the DSIP for brain stroke prophylaxis.     

c-fos Gene expression during emotional stress in rats: blocking by delta sleep-inducing peptide

An emotional stress induces an obvious immediate early gene c-fos expression in the brain limbic structures in the rats predisposed to emotional stress. Administration of the delta-sleep-inducing peptide (DSIP) was shown to inhibit the c-fos expression. It led to an obvious inhibition of the c-fos expression in paraventricular nuclei of the hypothalamus, medial and lateral parts of the septum of rats predisposed to emotional stress. This mechanism seems to play an important role in the DSIP anti-stress effects.     

The effect of the delta-sleep peptide on the adrenaline level in rat tissues under normal conditions and during cold stress

Adrenalin content in the brain, liver and adrenal glands under the effect of cold stress grows by 314, 500 and 56% as compared to the control. A single administration of the delta-sleep inducing peptide (DSIP) in a dose of 12 microgram/100 g to intact animals makes the adrenalin content in the brain higher 1, 3, 6 and 24h after administration; two and three days later the adrenalin content in the brain does not change. The amount of adrenalin in the liver of the same animals increases 1, 3, 6 h and 1, 2, 3 days after DSIP administration. Intraperitoneal administration of DSIP induces an increase of the adrenalin level in the adrenal glands of rats an hour and a day after administration. Two days later the level of adrenaline decreases by 41%; 3, 6 h and 3 days after DSIP administration the content of adrenaline remains unchanged. As a result of the DSIP administration in a dose of 12 micrograms/100 g to the animals in the state of cold stress, the content of adrenalin increases in the rat brain by 129, in the liver--by 300, adrenal glands--by 44% as compared with the control.     

Muramyl dipeptide-elicited production of PGD2 from astrocytes in culture

We used primary cultures of rat brain astroglial cells in order to investigate the interrelationship between PGD2 and other sleep-promoting substances such as muramyl dipeptide (MDP), lipopolysaccharide (LPS), delta-sleep-inducing peptide (DSIP), uridine, and interleukin 1 (IL-1). A large amount of PGD2 was released into the culture medium by stimulation with MDP, LPS, and IL-1 but DSIP and uridine failed to stimulate such release. These results suggest that PGD2 may be part of the series of biochemical steps involved in induction of sleep by MDP, LPS, and IL-1.     

Antiradical and antioxidant effect of arginine and its action on lipid peroxidation in hypoxia

Arginine (0.5 and 1 mu mol) decreased by 14-17% rates of superoxide anion formation in two systems: HADH-phenazine methosulfate and hydroxylamine autooxidation with nitrotetrazolium blue. These concentrations of arginine decreased by 40-50% formation of diene conjugates and Schiff bases in plasma when lipid peroxidation initiated by iron -ascorbate. Arginine administration before hypoxia decreased lipid peroxidation rate in plasma and liver microsomal membranes of rats.     

Action of the "delta-sleep peptide" on monoamine oxidase and acetylcholinesterase activity in subcellular fractions from different rabbit brain formations in vivo

Administration of delta-sleep-inducing peptide (DSIP) in vivo in a dose of 30 microgram/kg bw brings about MAO-A (substrate-serotonin) activation in synaptosome subfractions and cellular mitochondria from the brain structures (motor cortex, nucleus caudatus, thalamus). Activity of MAO-B (substrate-p-nitrophenylethylamine) and acetylcholinesterase was inhibited negligibly and specifically in subcellular fractions of the test brain structures. The results suggest that DSIP effects the regulatory or modulation function in the synapse. As one of the elements of sleep mechanisms this peptide induces a number of processes, particularly in serotonin metabolism.     

Appearance of parkinsonian syndrome after administration of delta sleep-inducing peptide into the rat substantia nigra

Bilateral microinjection of delta-sleep-inducing peptide (DSIP) (10.0 nmol) into the substantia nigra provoked hypokinesia and rigidity in rats observed during 4.0 hours. Injection of DSIP in dose of 5.0 nmol into the substantia nigra or into the nuclei caudati in dose of 10.0 nmol did not induce such symptoms. The enhanced slow-wave activity was recorded in caudate nuclei during hypokinesia and rigidity which demonstrated the formation of the generator of pathologically enhanced excitation (GPEE). The systemically cyclodol administration resulted in abolishment of rigidity and increase in locomotor activity. The conclusion is that bilateral intranigral DSIP injection caused acute parkinson syndrome in rats due to the formation of cholinergic GPEE in caudate nuclei. The hyperactive caudate nuclei act as the pathologic determinant which induces the parkinson syndrome.     

Age-related changes in the activity of free-radical processes in rat tissues and blood serum]

Activity of Cu,Zn-superoxide dismutase (SOD) in old rats' brain was found to be decreased by 46.8% as compared to young animals. The brain concentration of Schiff bases (SB) was decreased by 13.6% in old rats, whereas concentration of diene conjugates (DC), protein peroxidation (PP), and total antioxidative activity (TAA) was the same in old as well as in young rats. The liver level of the DC and TAA was also the same. The serum level of the PP, SB, and DC was increased whereas the activity of the SOD and TAA was decreased in old rats. The findings suggest occurrence of considerable age-related changes in free radical processes as well as the organ specifics of these changes in rats.     

Metabolic features of the adaptive effect of delta-sleep inducing peptide and piracetam under hyperoxic conditions.

Adaptive effects of delta-sleep inducing peptide (DSIP, 12 microgram/100 g body weight, single intraperitoneal injection) and piracetam (3 mg/100 g body weight, daily intraperitoneal injection for 3 days) are manifested via differential changes in neurotransmitter amino acids (GABA, glutamate, aspartate), modulation of transport ATPase activity, and decreased accumulation of lipid peroxidation products (conjugated dienes, malonic dialdehyde, Schiff bases) in various fractions of neuronal membranes (myelin, synaptic and mitochondrial membranes) in the sensomotor cortex of rat brain. Under hyperbaric oxygenation (0.3 MPa for 2 h), the combination of DSIP and piracetam enhanced the protective effect of each compound.     

Relation between the parameters of lipid peroxidation and the constant potential of the rat brain

Studies were carried out of the relationship between the constant potential level (CPL) and the intensity of lipid peroxidation (LP) of the brain membranes in rats with different motor asymmetry. Strong correlation was found between CPL and the content of LP products (diene conjugates, hydroperoxides and Schiff bases) in the hemispheres of the brain (R = 0.7-0.8).     

Antihypoxic and antioxidative properties of bemitil

The authors discovered antihypoxic properties of the bemitil (pretreatment injections 50 mg/kg intraperitoneally) in the experiments on rats with the circulatory or hypoxic hypoxia. There was limitation of pO decrease and diene conjugates and Schiff bases production increase with the drug in the circulatory hypoxia conditions. Bemitil restricted malondialdehyde accumulation in the rat brain homogenate under the activation of free radicals processes. In the mitochondrial suspension incubation similar effect of the medicine was accompanied with limitation of organelle degradation. Bemitil showed no antiradical activity.     

Lipid peroxidation rate and the antioxidant protection system during a three-day dry immersion experiment

The content of lipid peroxidation products (diene conjugates, malondialdehyde, Schiff bases) and antioxidant defense system indices (the main lipid antioxidant tocopherol and the level of general antioxidant activity) were measured in the blood serum of five male volunteers aged 25?C40 years in a three-day dry immersion experiment. During the immersion test, no deviations of indices from the background values were found. An increase in the tocopherol concentration within 2 h after the beginning of the experiment was the only exception. A significant increase in the concentration of lipid peroxidation products, particularly, diene conjugates, was observed 2 h after immersion completion during the reconditioning period. However, the tocopherol content was significantly lower than the background values. It is concluded that the subjects?? adaptation to simulated microgravity conditions displays no pronounced stress component, whereas bringing back to normal vital functions after exposure to immersion induces a pronounced stress reaction illustrated by a significant increase in the lipid peroxidation product levels against a background of a decrease in the functional activity of the antioxidant defense system.     

The effect of naloxone and delta-sleep inducing peptide on the homocarnosine content of brain tissue

The intraventricular and intravenous administration of naloxone was studied for its effect on the homocarnosine amount in cerebral hemispheres, striatum, hippocamp, hypothalamus, thalamus, cerebellum, medulla oblongata as well as in the spinal cord of rabbits. The intracysternal administration of naloxone decreases the homocarnosine amount in the striatum, hypothalamus, cerebellum and medulla oblongata. The intravenous administration of peptide exerts no statistically reliable effect on the homocarnosine content in the rabbit brain. The intraperitoneal administration of delta-sleep-inducing peptide increases sharply the homocarnosine content in the rat brain.     

Effect of leu-enkephalin and the delta-sleep-inducing peptide, M (DSIP), on endogenous noradrenaline release by brain synaptosomes in the rat

Fluorometry was employed to measure the noradrenaline (NA) content in rat brain synaptosomes depending on the duration of incubation, depolarization effects (40 mM KCl or 1.5 mM ouabain), composition of the synaptosomal fraction and concentration of the peptides. The 10-minute incubation in a potassium medium of a suspension of light synaptosomes was used as an optimal test-system for studying the peptide action. Leu-enkephalin inhibited the depolarization-induced NA release. The effect was abolished by naloxone. The delta-sleep-inducing peptide (DSIP) did not influence the neurotransmitter release at concentrations of 10(-8)-10(-5) M. A mixture of amino acids imitating the amino acid composition of the DSIP influenced spontaneous release of NA. This effect is discussed in connection with the physiological action of the peptide on its intraventricular injection.     

Level of delta sleep-inducing peptide (DSIP) in the brain of rats with different alcoholic motivation

Radioimmunoassay was used to measure the content of delta-sleep-inducing peptide (DSIP) in random-bred albino rats divided into groups according to the duration of ethanol anesthesia and the levels of 15% ethanol consumption under free-choice conditions. The concentration of the neuropeptide was assayed in intact brain, in the cortex of large hemispheres, medulla oblongata, thalamus and striatum. The short-sleeping rats manifested a statistically significant lowering of the DSIP content in intact brain homogenates, in the cortex of large hemispheres and striatum. On the contrary, thirty minutes after a single intraperitoneal injection of ethanol in a dose of 1 g/kg the DSIP content in the medulla oblongata, thalamus and striatum was found to be increased. The raising of the ethanol dose up to 2.5 and 4.5 g/kg was followed by a less significant increase in the neuropeptide content. Prolonged chronic alcoholization under free-choice conditions led after 12 months to the reduced DSIP content in the medulla oblongata, thalamus and striatum. The importance of DSIP for the pathogenesis of experimental alcoholism using rats with different levels of alcoholic motivation is discussed.     

Delta-sleep-inducing peptide (DSIP): An update

The isolation and characterization of delta-sleep-inducing peptide (DSIP) achieved from 1963 to 1977 were reviewed in 1984. The first reports describing sleep as well as extra-sleep effects of DSIP also were included in that work. Only two years later, much additional literature concerning DSIP has accumulated. Besides further sleep-inducing and/or-supporting effects of DSIP in animals, considerable work has been carried out to evaluate the potential use of the peptide for therapeutic purposes such as treatment of insomnia, pain, and withdrawal. Immunohistochemical as well as radioimmunochemical studies provided further insights into the natural occurrence of the nonapeptide and the distribution of DSIP-like material in the body, suggesting possible relations of the peptide to certain diseases. Various physiological functions of DSIP and a possible mechanism of action involving the modulation of adrenergic transmission remain to be established.