Synthetic corticotropins and the GABA-receptor system: Direct and delayed effects.

The central effectors of the stress system are greatly interconnected and include, among others, a large group of peptides derived from proopiomelanocortin. In addition to natural corticotropins, a number of artificial molecules that contain some ACTH fragments in their structure are also referred to members of this family. Some of them possess a wide range of biological activity. The molecular mechanism underlying the biological activity of such peptides is partly based on allosteric modulation of various receptors. We analyzed the ability of some biologically active synthetic corticotropins (ACTH(4-7)PGP, ACTH(6-9)PGP, ACTH(7-10)PGP), and glyproline PGPL to affect the GABA-receptor system of rat brain. The effects of the peptides were studied in the isolated plasma membranes of brain cells, as well as after systemic peptide administration in the rat model of acute restraint stress. The delayed effect of stress or preadministration of each of the studied peptides on [3 H]GABA binding was different for its high- and low-affinity-specific sites. The studied peptides individually affected the binding of [3 H]GABA in their own way. Acute restraint stress caused a decrease in [3 H]GABA binding at its low-affine site and did not affected the high-affine site. Preliminary peptide administration did not influence this effect of stress.

α1-Adrenergic receptor regulates papillary muscle and aortic segment contractile function via modulation of store-operated Ca2+ entry in long-tailed ground squirrels Urocitellus undulatus.

The effect of phenylephrine (PE) on right ventricle papillary muscle (PM) and aortic segment (AS) contractile activity was studied in long-tailed ground squirrels Urocitellus undulatus during summer activity, torpor and interbout active (IBA) periods in comparison to rat. We found that PE (10 µM) exerts positive inotropic effect on ground squirrel PM that was blocked by α1-AR inhibitor-prazosin. PE differently affected frequency dependence of PM contraction in ground squirrels and rats. PE significantly increased the force of PM contraction in summer and hibernating ground squirrels including both torpor and IBA predominantly at the range of low stimulation frequencies (0.003-0.1 Hz), while in rat PM it was evident only at high stimulation frequency range (0.2-1.0 Hz). Further, it was found that PE vasoconstrictor effect on AS contractility is significantly higher in ground squirrels of torpid state compared to IBA and summer periods. Overall vasoconstrictor effect of PE was significantly higher in AS of ground squirrels of all periods compared to rats. Positive inotropic effect of PE on PM along with its vasoconstrictor effect on AS of ground squirrels was not affected by pretreatment with inhibitors of L-type Ca2+ channels, or Na+/Ca2+ exchanger or Ca2+-ATPase but was completely blocked by an inhibitor of store-operated Ca2+ entry (SOCE)-2-APB, suggesting the involvement of SOCE in the mechanisms underlying PE action on ground squirrel cardiovascular system. Obtained results support an idea about the significant role of alpha1-AR in adaptive mechanisms critical for the maintaining of cardiovascular contractile function in long-tailed ground squirrel Urocitellus undulatus.

ACTH(6-9)PGP Peptide Protects SH-SY5Y Cells from H2O2, tert-Butyl Hydroperoxide, and Cyanide Cytotoxicity via Stimulation of Proliferation and Induction of Prosurvival-Related Genes.

Stabilized melanocortin analog peptide ACTH(6-9)PGP (HFRWPGP) possesses a wide range of neuroprotective activities. However, its mechanism of action remains poorly understood. In this paper, we present a study of the proproliferative and cytoprotective activity of the adrenocorticotropic hormone fragment 6-9 (HFRW) linked with the peptide prolyine-glycyl-proline on the SH-SY5Y cells in the model of oxidative stress-related toxicity. The peptide dose-dependently protected cells from H2O2, tert-butyl hydroperoxide, and KCN and demonstrated proproliferative activity. The mechanism of its action was the modulation of proliferation-related NF-κB genes and stimulation of prosurvival NRF2-gene-related pathway, as well as a decrease in apoptosis.

Semax, synthetic ACTH(4-10) analogue, attenuates behavioural and neurochemical alterations following early-life fluvoxamine exposure in white rats.

Selective serotonin reuptake inhibitors (SSRI) are commonly used to treat depression during pregnancy. SSRIs cross the placenta and may influence the maturation of the foetal brain. Clinical and preclinical findings suggest long-term consequences of SSRI perinatal exposure for the offspring. The mechanisms of SSRI effects on developing brain remain largely unknown and there are no directional approaches for prevention of the consequences of maternal SSRI treatment during pregnancy. The heptapeptide Semax (MEHFPGP) is a synthetic analogue of ACTH(4-10) which exerts marked nootropic and neuroprotective activities. The aim of the present study was to investigate the long-term effects of neonatal exposure to the SSRI fluvoxamine (FA) in white rats. Additionally, the study examined the potential for Semax to prevent the negative consequences of neonatal FA exposure. Rat pups received FA or vehicle injections on postnatal days 1-14, a time period equivalent to 27-40 weeks of human foetal age. After FA treatment, rats were administered with Semax or vehicle on postnatal days 15-28. During the 2nd month of life, the rats underwent behavioural testing, and monoamine levels in brain structures were measured. It was shown that neonatal FA exposure leads to the impaired emotional response to stress and novelty and delayed acquisition of food-motivated maze task in adolescent and young adult rats. Furthermore, FA exposure induced alterations in the monoamine levels in brains of 1- and 2- month-old rats. Semax administration reduced the anxiety-like behaviour, improved learning abilities and normalized the levels of brain biogenic amines impaired by the FA exposure. The results demonstrate that early-life FA exposure in rat pups produces long-term disturbances in their anxiety-related behaviour, learning abilities, and brain monoamines content. Semax exerts a favourable effect on behaviour and biogenic amine system of rats exposed to the antidepressant. Thus, peptide Semax can prevent behavioural deficits caused by altered 5-HT levels during development.

A new derivative of acetylsalicylic acid and carnosine: synthesis, physical and chemical properties, biological activity.

PURPOSE: The aim of this study was to create and assess biological activity of a new compound based on carnosine and acetylsalicylic acid (ASA) that will comprise antioxidant effect with antiplatelet activity, while simultaneously preventing side effects on the gastrointestinal tract. METHODS: Salicyl-carnosine (SC) was synthesized by condensation of ASA and carnosine. Antioxidant activity was determined by spectrophotometric and chemiluminescence methods. Antiplatelet activity was carried out by the light transmission-aggregometry method using the inductor ADP. Chronic gastric ulcer in rats was modeled using glacial acetic acid. RESULTS: Using SOD-like activity, iron-induced chemiluminescence, BaSO4-activated respiratory burst, and evaluation of red blood cell structure stabilization during oxidative damage induced by sodium hypochlorite, it was shown that SC possesses antioxidant activity analogous, or better, than that of carnosine. Antiplatelet activity of SC was evaluated in the blood of healthy individuals, and was also shown to be comparable to, or exceeding that of ASA. Also SC demonstrates high resistance to hydrolysis by tissue and serum carnosinases. Most importantly, it was shown that SC has protected the gastric mucosa against the formation of stomach ulcerative lesions and promoted their epithelization, therefore overcoming the undesirable inherent side effects of ASA. CONCLUSIONS: SC preserves pharmacologically significant properties of ASA and carnosine while retaining an anti-ulcer activity and resistance to the carnosinase hydrolysis at the same time. These properties are particularly promising for the potential development of new anti-inflammatory and antithrombotic drugs. Graphical abstract .

Mechanisms for glyproline protection in hypercholesterolemia.

Comparative analysis of the hypocholesterolemic and antithrombotic action of small regulatory glyproline peptides (Pro-Gly-Pro, Arg-Pro-Gly-Pro and Pro-Gly-Pro-Leu) was performed on an experimental hypercholesterolemia model of rats. Repeated intranasal introduction of glyproline peptides to fat-diet-fed animals led to more active functioning of the anticoagulation system (the anticoagulant and fibrinolytic properties of the plasma increased and platelet aggregation decreased) and to normalization of the total cholesterol level as a parameter of lipid metabolism. The largest anticoagulant and hypocholesterolemic effect was detected for the Pro-Gly-Pro-Leu peptide. Hypothetical mechanisms of antithrombotic and hypocholesterolemic effects of glyproline peptides are presented.

Effects of behaviorally active ACTH (4-10) analogue - Semax on rat basal forebrain cholinergic neurons.

PURPOSE: It is well established that cholinergic neurons of the basal forebrain degenerate in Alzheimer's dementia. Although recent studies were concentrated on screening molecules that might reduce the concomitant cell loss, little is known about therapeutically promising molecules. We studied the effect of Semax (Met-Glu-His-Phe-Pro-Gly-Pro), a behaviorally active adrenocorticotropic hormone (4-10) analogue, on survival of cholinergic basal forebrain neurons in vitro. Semax is known to stimulate learning and memory and can be successfully used for treatment of ischemic stroke. METHODS: Primary cultures of neuronal and glial cells from basal forebrain of rats were used in all experiments. The stability of Semax in cell cultures was tested by HPLC analysis. Cell survival in neuronal cultures was quantitated using immocytochemical and cytochemical analyses as well as detection of choline acetyltransferase activity. RESULTS: We have shown that Semax may approximately 1.5-1.7 fold increase survival of cholinergic basal forebrain neurons in vitro. Moreover, Semax (100 nM) stimulated activity of choline acetyltransferase in dissociated basal forebrain tissue cultures. However, the numbers of GABA-ergic neurons, total neuron specific enolase neurons were not affected. In concentration from 1 nM to 10 microM, Semax did not affect proliferation of glial cells in primary cultures. CONCLUSION: Implications of these findings with respect to Alzheimer's disease remain to be clarified.

The seasonal peculiarities of force-frequency relationships in active ground squirrel Spermophilus undulatus ventricle.

The plasticity of calcium homeostasis is of crucial importance for the unique ability of the hibernators' heart to function under conditions of body temperature changing from 37 degrees C to near freezing point. However, the precise mechanism of calcium homeostasis regulation in these animals is largely unknown. Force-frequency relationship, as an indicator of participation of various sources of calcium (external and intracellular) in the activation of contraction, and post-rest potentiation as an index of the capacity of sarcoplasmic reticulum (intracellular calcium source) to store and release Ca(2+), were studied to analyse the role of different calcium-transporting systems in seasonal and temperature-induced changes in isometric twitch force of ground squirrel papillary muscles. The obtained results revealed significant functional differences during the annual cycle, which are indicative of an increased role of the sarcoplasmic reticulum in regulation of contractility in animals in transition to the hibernation period. Also, how myocardium during the hibernation period copes functionally with acute decreases in temperature was investigated.

Long-lasting changes of albino rats behavior and brain bioamines content after immunization against cholecystokinin-3 and -4.

Active immunization against cholecystokinin fragments 31-33 (CCK-3) and 30-33 (CCK-4) results in long-lasting changes of albino rats' behavior. CCK-3 and CCK-4 covalently linked to antigen-carrier evokes the suppression of the anxiety, decreases some signs of depression-like behavior and changes the level of bioamines and their catabolites in striatum at least for two months after immunization. These data can provide a perspective approach to the problem of long term correction of behavior.