Effect of dalargin on the processes of memory and learning in the rat

In experiments on rats, the influence was studied of dalargin on the elaboration and preservation of various homogeneous and heterogeneous conditioned reflexes (CRs) elaborated in single and multiple pairings. The effect of dalargin on the processes of learning and memory was compared with the action of the peptide on the activity of hypothalamic neurones. Administration of dalargin delayed the elaboration of maze defensive CRs and practically did not affect the elaboration of two-way avoidance. The preservation of CR also deteriorated under the influence of dalargin. Administration of dalargin 10 min before the CRs testing did not prevent their reproduction. When using CRs elaborated in a single pairing, dalargin disturbed the preservation of the drinking CR and improved that of passive avoidance CR. Dalargin in this dose affected the emotional state of animals in the open field and did not significantly affect their motor activity. Dalargin suppressed impulse activity in 17 out of 22 tested neurones of the lateral hypothalamus, with maximum effect in 20-50 min after its administration. The obtained data show that the character of dalargin action on the elaboration of CR and mainly on its consolidation, depends on the character of the elaborated CR and is probably due to great extent to the effect of the peptide on the brain emotional mechanisms.     

Effect of opioid peptides on the lymphatic drainage of the pancreas in the rat and dog

The effect of dalargin and some other ligands of the opioid receptors on drainage function of the pancreatic lymphatic system was studied in rats and dogs. In rats, dalargin (30-1000 micrograms/kg, subcutaneously) accelerated the elimination of Evans blue from beneath the pancreatic capsule in a dose-related manner. The effect of dalargin was attenuated by naloxone. Specific agonists of mu-, delta- and sigma-opioid receptors had no dalargin-like activity. In dogs, dalargin (60-80 micrograms/kg, subcutaneously) after the administration of Evans blue into the pancreas increased its concentration in the truncus lymphaticus and slowed down its penetration into the blood. Thus, dalargin accelerates the elimination of Evans blue from the pancreas due to the intensification of lymphatic drainage. The effect of dalargin was mediated by subpopulation of opioid receptors with which their certain selective ligands have but slight interaction.     

Molecular mechanisms of antioxidant action of dalargin on the liver in experimental cholestasis]

Changes of antioxidant activity of dalargin in the liver after naloxone (100 micrograms/kg) administration were examined in experiment on 144 rats with cholestasis. It was found that dalargin inhibited the activity of xanthine oxidase by 32-37% in different time periods after the injection. Dalargin and naloxone, when used in combination, had no effect on the enzyme activity. Glutathione-S-transferase activity rose by 38.0% and 21.8% on hour 1 and 3 after the injection, respectively, while simultaneous injection of dalargin and naloxone induced no changes in the enzyme activity after 1 hour, though decreased it by 36.8% and 26.4% on hour 3 and 5, respectively. Dalargin inhibited lipid peroxidation by 29-35%, simultaneous injection of dalargin and naloxone raised lipid peroxidation by 109.2%, 80.7% and 25.7% after 1, 3 and 5 hours, respectively. Dalargin injection elucidated a marked tendency to lowering of blood release of the liver-specific enzymes histidase and urokaninase in line with enhancement of their activity in the liver. A combined injection of dalargin and naloxone promoted high release of histidase and urokaninase in blood and did not change histidase activity in the liver in all cases. Urokanidase activity elevated in 5 hours. It was noticed that dalargin raised leu-enkephalin levels in the liver 3.5-fold 1 h after the injection. The reduced dalargin antioxidant effect coupled with naloxone pretreatment demonstrated indirect action of the neuropeptide on the liver via neuron receptors of the liver.     

Effects of several regulatory peptides on the functional activity of the pancreas in acute experimental pancreatitis

The influence of regulatory peptides (somatostatin, calcitonin, and dalargin) on xanthine oxidase activity and lipid peroxidation level in pancreatic tissues as well as on the release of pancreatic enzymes (alpha-amylase, trypsin, lipase, and transamidinase) into blood was studied in 205 rats with experimental acute pancreatitis. Somatostatin and dalargin were shown to have obvious antioxidant effect seen by reduced xanthine oxidase activity and MDA level. All studied peptides stimulate reduced release of pancreatic enzymes. Particularly, reduction of dalargin and somatostatin is caused by inhibition of their synthesis as well as by pancreas protective effect of the peptides. Release of enzymes reduced by calcitonin is probably associated only with inhibition of secretory activity of the pancreas.     

Effect of the hexapeptide dalargin on ornithine decarboxylase activity in the duodenal mucosa of rats in experimental duodenal ulcer

The effect of an opioid antiulcerogenic hexapeptide dalargin on ornithine decarboxylase activity of duodenal mucosa has been studied in rats with experimental duodenal ulcers induced by cysteamine. The intraperitoneal injection of 12.5 micrograms/kg of dalargin inhibited ulcerogenesis and activated the enzyme. The effect of the peptide was antagonized by an opiate antagonist naloxone. 5000 micrograms/kg of dalargin failed to inhibit the ulcer formation or to activate ornithine decarboxylase. Since ornithine decarboxylase activation is a marker of intensified cell proliferation and tissue regeneration, our results suggest that the antiulcerogenic effect of dalargin is due to the enhancement of duodenal mucosa regeneration.     

Ultrastructure of cardiomyocytes in the peri-infarct zone during the treatment of experimental myocardial infarct in rats using the hexapeptide dalargin

The experiments on white rats with induced myocardial infarction have studied the influence of dalargin on the infarction size and peri-infarction zone ultrastructure. 24 hours later the decrease in the infarction zone size was detected in rats who had received dalargin in a dose of 50 and 100 micrograms/kg. In the peri-infarction zone the increase in glycogen quantity, the lower degree of lipid infiltration, the increase in mitochondrial number and mitochondrial energy effectiveness coefficient were noted, as compared to control animals. Sarcolemma of cardiomyocytes from the peri-infarction zone in rats on dalargin was impermeable for colloidal lanthanum. The decrease in the infarction size under the effect of dalargin is explained by its influence on the survival of cardiomyocytes in the peri-infarction zone.     

Pharmacological correction of experimental alcoholic embryopathy

In the experiments on the progeny of ethanol-exposed rats it was shown that consumption of 15% alcohol instead of water during pregnancy resulted in the worsening of shuttlebox avoidance learning and decrease in succinate dehydrogenase activity in the visual and sensorimotor cortex. These data are indicative of cerebral hypoxia during embryogenesis. The injection of synthetic opioid peptide dalargin during critical periods of development (at the end of embryogenesis and early ontogeny) prevented partially the disturbances of higher nervous activity and tissue breathing which were induced by alcohol intoxication. Possible mechanisms of positive dalargin effect are discussed.     

Effects of opioid receptor ligands on DNA synthesis and histamine contents in the gastric mucosa and blood of white rats

Using autoradiographic study with 3H-thymidine and spectrofluorometric method the authors studied the influence of opioid receptors' ligands on the DNA synthesis in the stomach epithelium histamine content and the blood. Leu-encephalin, B-endorphin, dalargin, napoxone were administered intraperitoneally to male rats. The dose of injection was 0.1 ml per 100 g body weight. It was observed that B-endorphin and dalargin 1.5-1.6 fold increased the quantity of DNA-synthesised nuclei in the epithelium of fundal stomach section. Leu-encephalin and dalargin increased the histamine concentration in the stomach, at the same time dalargin caused a rapid decrease of histamine concentration in the blood. Naloxone also decreased histamine concentration in the stomach. The obtained results are being discussed in connection with dalargin therapy of ulcerous diseases.     

The noradrenaline and serotonin content in symmetrical parts of the normal rat brain and during learning and peptide administration]

Content was compared of noradrenaline (NA) and serotonine (5-OT) in the right and left halves of the rats brain in norm, at elaboration of defensive conditioned reflexes of two-ways avoidance (CRTWA) and at administration of neuropeptides influencing the learning and memory--dezglycilargininvasopressin (DG-AVP), ACTH4-7 pro-gli-pro and dalargin. The conducted studies showed that in control animals the content of NA in the cortex of the right hemisphere was significantly higher than in the cortex of the left one. For the content of 5-OT in symmetric brain parts no significant differences were revealed. Under the elaboration of CRTWA the asymmetry of NA content was not eliminated. Systemic administration of DG-AVP, ACTG4-7 pro-gli-pro and dalargin practically did not change the content of 5-OT, but reduced the content of NA in the cortex and the rest of the brain, and the content of NA in the right and left cortex was equalized. The obtained data point to the asymmetric character of neuropeptides action and to greater resistance of 5-OT-ergic brain system to functional load and to administration of peptides in comparison with NA-ergic system.     

Comparative effect of opioid receptor ligands on cell division in the epithelium of the tongue in white rats

Beta-endorphin, leu-enkephalin, dalargin and naloxone influences on cell division have been studied in tongue epithelium of white rats. The preparations were administered at a dose of 0.1 ml per 100 g body weight as a 2.10(-9) M solution. Cell division was studied 24 hours after administration. beta-endorphin, leu-enkephalin, dalargin and naloxone caused a 1.5-1.7-fold increase in the number of DNA-synthesizing nuclei, which was accompanied by an adequate rise in mitotic index in experiments with dalargin.     

The use of 1H-NMR-spectroscopy for the study of peptide degradation by angiotensin-converting enzyme

A new method for continuous registration of enzymatic hydrolysis of peptides involving 1H-NMR spectroscopy was developed. The advantages of the method were demonstrated, using dalargin (Tyr-D-Ala-Gly-Phe-Leu-Arg) hydrolysis catalyzed by human kidney angiotensin-converting enzyme as an example. It was shown that the maximal activity of the enzyme towards dalargin is observed at pH 7.8; Km is 0.35 mM. The enzyme is inhibited by the substrate (Kd = 0.55 mM). Cl- do not influence the catalytic activity of the enzyme with respect to dalargin. The stereospecificity of the angiotensin-converting enzyme towards dalargin diasteriomers was studied.     

Effects of dalargin on proliferative processes and vertical migration of cells of the corneal epithelium during stress

We studied the effect of synthetic analogue of opioid peptides dalargin on kinetics of cell population of corneal epithelium during stress. It was found out that dalargin introduction before stress decreased the level of pathological mitoses induced by the acute stress action in white rats early in the morning. Dalargin prevented the activation of the proliferative process after the repeated stress activation, made stress milder. The stress induced vertical migration of the cells of the corneal epithelium normalized by dalargin. We believe these results show that dalargin has a good protective effect.     

Individual typologic characteristics of the open-field behavior of rats

By means of ethograms record and analysis, connection has been studied between the properties of rats behaviour organization in the open field, determining the level of behaviour entropy in this test, and the speed of conditioned reflexes formation in the Skinner chamber. According to behaviour entropy level the rats are significantly divided into four groups; the lowest speed of conditioned reflexes formation in the Skinner's chamber is observed in the animals of the first (low entropy) group, the highest--in the fourth (high entropy) group. The obtained data are discussed according to Pavlov's concepts on the characteristics of the basic nervous processes, determining individual-typological characteristics of the higher nervous activity of the animal. Conclusion is made that division according to the level of behaviour entropy in the open field test may serve as a safe method of express-estimation of the animals abilities to conditioned habits formation.     

Peptide therapy of sleep disorders in neurotic rats

As a result of chronic stress, anxiety appeared in the rats behaviour, motor activity increased, heart rate quickened, blood pressure raised, conditioned instrumental alimentary reflexes missed, the duration of deep phases of sleep lowered, time of falling asleep became longer, the number of awakening increased. The change in quantitative characteristics of sleep was accompanied by its worsening, especially of rapid sleep. Administration of substance P (SP) eledoisin hexopeptide (EH) (250 mcg/kg), 100-200 mcg/kg of delta sleep peptide and 10 mcg/kg of ethylcrotyl barbiturate improved the rats behaviour and sleep parameters. Calipnon accelerated falling asleep. Delta sleep peptide increased sleep duration. SP and EH restored not only the quantitative characteristics of deep phases of sleep but greatly improved their quality: lowered the blood pressure disrupted tachycardia, normalized the conditioned activity.     

Features of the formation and dynamics of short-term image memory in the rat

Direct and indirect methods of delayed reactions showed that short-term image memory of rats elaborated on the basis of conditioned signals in poorly developed and its duration is not more than 1 s. While testing this memory, changes in the behaviour were seen, pointing to the development of "hard state" in the higher nervous activity of animals. Short-term image memory based on complex perception was shown to be well developed in rats, and by many characteristics it approximated to the memory of highly organized animals. The ability to reproduce the short-term image memory on the basis of conditioned signals is supposed to be a complex form of psychonervous activity of animals, and in rodents it appears to be at an initial stage of development.     

Effect of a stable analog of leu-enkephalin, dalargin, on the cell division of the corneal epithelium in white rats

The effect of Leu-enkephalin analog--dalargin--on the corneal epithelium proliferation has been studied in white rats. 10 microliter dalargin per 1 kg body weight were administered intraperitoneally at 8 a.m. The mitotic index (MI), DNA synthesis cell index and label intensity (LI) were determined every 4 hours over a 24-hour period. The results obtained demonstrate that dalargin stimulates DNA synthesis in cells throughout the entire period of action. MI increased only 4, 8, 12 hours after dalargin administration. Mean daily DNA synthesis cell index and MI increased 2.1-fold and 3.1-fold, respectively after dalargin administration. It is suggested that dalargin activates the cell division processes by speeding up mitosis, shortening the premitotic period, accelerating the speed of the DNA synthesis and increasing cell proliferation pool.     

Effects of dalargin on hemodynamics of anesthesized rats during truncal vagotomy

I/v dalargin injection (20-25 g/kg) to narcotized rats in case of total myoplegia effectively protect hemodynamic changes under nociceptive stimulation. Bilateral truncal vagotomy partially decrease the protective effect of dalargin. Protective effect of the medicine results in activation of central and peripheral opioid receptors.     

Effect of microinjections of a selective blocker of M1-muscarinic receptors pirenzepine into the neostriatum on the rat motor activity

In simulated discrimination conditioned reflex of active avoidance (CRAA) in T-maze, the effect of bilateral microinjections of the muscarinic receptor M1 selective blocker pirenzepine on the CRAA formation and behaviour in the "open filed" test, was studied in rats. A sharp worsening of the CRAA learning and a significant increase in the motor activity were shown to occur in rats following the microinjections as compared with control rats. The change in the motor responses seems to account for the worsening of the CRAA learning. Another reason of the phenomenon could involve a disorder in perception of conditioned signals and their poor differentiation. The data obtained and the literature data suggest a complex character of changes induced by the blockade of the M1 muscarinic receptors of the neostriatum.     

Effect of the local administration of 5,7-DHT and 6-OHDA into the neocortex on the learning and exploratory behavior of rats in an open field

On Wistar rats characteristics were studied of investigating behaviour in the open field, of learning of conditioned food-reinforced reaction and also of BA and their metabolites content in various brain structures under local intracerebral injections of specific neurotoxins; 6-hydroxydopamine (6-OHDA) and 5,7-dihydroxytryptamine (5,7-DHT), abolishing correspondingly catecholaminergic and serotoninergic terminals. Bilateral injection of 6-OHDA in the neocortex led to a weakening of rats investigating activity in the open field and to an increase of the time of fulfillment of the forming of conditioned food-reinforced reaction. Administration of 5,7-DHT was accompanied by an increase of the investigating behaviour in the open field and a reduction of the duration of the forming of conditioned reaction. Administration of 6-OHDA to the neocortex caused a lowering of catecholamines level in the frontal area of the neocortex and the hippocampus. Analogous administration of 5,7-DHT elicited simultaneously with a deep level lowering of 5-HT and its metabolite in these structures, a change of catecholamines content which testifies to a lesser specificity of the neurotoxin 5,7-DHT in comparison with 6-OHDA. Structures lesion of serotoninergic and catecholaminergic systems of the frontal cortex and the hippocampus brought about by a local administration of 6-OHDA and 5,7-DHT in the neocortex was accompanied by differently directed changes in animals behaviour.     

The characteristics of higher nervous activity and of the brain benzodiazepine system in rats subjected to ethanol exposure in the prenatal period

Exposure to ethanol during pregnancy results in the alternation of 3H-diazepam binding to synaptosomal neocortical membranes from the rat offspring. In male experimental rats, 14 days of age, binding level diminished to 11%. In two-month-old control rats Scatchard plot was biphasic. It has been shown that prenatal exposure to ethanol leads to changes in the nature of binding in two-month-age experimental animals, as compared with the control ones. 3H-diazepam binding changes went along with behavioural deviations. In experimental rats locomotor activity was increased in the "open field" test, passive avoidance conditioned reflex retention was decreased and elaboration parameters of active avoidance conditioned reflex were changed, as compared with the control ones. The data obtained show that higher integrative functions were disturbed by prenatal alcoholization. Correlations between benzodiazepine receptor state and behaviour were studied.