Effect of Hydrogen Sulfide on Deformability of Rat Erythrocytes.

The influence of a hydrogen sulfide donor NaHS (2×10-5-10-3 M) on the rat erythrocyte deformability was analyzed by laser diffractometry. NaHS (6×10-5 M) increased, while NaHS (10-3 M) reduced erythrocyte deformability. The effect of NaHS (6×10-5 M) was similar to that of NO donor sodium nitroprusside (SNP, 10-7 M). However, simultaneous use of NaHS (6×10-5 M) and SNP induced less pronounced changes in erythrocyte deformability than their individual application. It is likely H2S, similar to NO, is involved in the regulation of erythrocyte deformability in the microvascular bed.

Changes in Sympathetic Innervation of the Heart in Rats with Experimental Myocardial Infarction. Effect of Semax.

The effect of peptide Semax on remodeling of cardiac sympathetic innervation was examined in rats with experimental myocardial infarction. In 28 days after ischemia/reperfusion injury, Semax diminished the growth of sympathetic innervation of ventricular septum, although it produced no effect on the density of ß1 and ß2 adrenoceptors.

[The role of ATP-dependent potassium channels and nitric oxide system in the neuroprotective effect of preconditioning]. / Rol' ATF-chuvstvitel'nykh kalievykh kanalov i oksida azota v zashchitnom effekte prekonditsionirovaniya mozga.

AIM: To study a role of ATP-dependent potassium channels (K+ATP) in the neuroprotective effect of ischemic (IP) and pharmacological (PP) preconditioning and evaluate the dynamics of blood nitric oxide (NO) metabolites in cerebral ischemia. MATERIAL AND METHODS: A model of ischemic stroke induced by the electrocoagulation of a middle cerebral artery (MCA) branch was used in male rats (n=86). Glibenclamide, a selective inhibitor of ATP-sensitive K+ channels, and diazoxide, a potassium channel activator, were used. IP and PP were performed 24 h before MCA occlusion. Blood concentrations of NO, NO3- and NO2-were measured 5, 24 and 72 h after occlusion. RESULTS: IP decreased a lesion area by 37% (p<0/05) and the preliminary introduction ofglibenclamide levelled the effect of IP. A protective effect of PP was similar to that of IP. A decrease in oxygenated R-conformers of Hb-NO and a reverse increase in non-oxygenated T-conformers as well as NO3- и NO2-were noted 5h after MCA occlusion. In the first 24 h after MCA occlusion, contents of NO3- and NO2- returned to normal values. There were changes in the concentrations of Hb-NO complexes as well, with the predominance of R-conformers and minimal contents of T-conformers. Moreover, the correlations between K+ATP channel blockade and the decrease in serum NO3- and NO2 were found (p<0/03). CONCLUSION: The neuroprotective effect of preconditioning is caused by the activation of K+ATP channels. An analysis of NO metabolite concentrations in the blood of rats with IP suggests that Hb-NO complexes belonging to R-conformers deposit and carry NO in tissues releasing NO accumulated via R→T transfer in conditions of ischemia.

Changes in Sympathetic Innervation of Rat Caudal Artery in Experimental Myocardial Infarction. Effect of Semax Peptide.

Activation of the sympathetic nervous system aggravates the course of myocardial infarction. Semax peptide moderated the degree of this activation and prevented the increase in the density of sympathetic endings in rat caudal artery in 28 days after ischemia or ischemia/reperfusion. The peptide reduced the density of α-adrenoreceptors in the caudal artery of rats with myocardial infarction. Semax produced no effect on ß-adrenoreceptors in both experimental models. The experiments on isolated segments of the caudal artery revealed reduced vascular responsiveness to electrical stimulation and norepinephrine infusion in rats treated with Semax after ischemia/reperfusion injury.

Effects of activated protein C on the size of modeled ischemic focus and morphometric parameters of neurons and neuroglia in its perifocal zone.

The effects of activated protein C (APC) on the quantitative parameters of neurons and neuroglia in the perifocal zone of infarction induced in the left hemispheric cortex were studied in two groups of rats. Group 1 animals served as control (control infarction). Group 2 rats were injected with APC (50 µg/kg) in the right lateral cerebral ventricle 3 h after infarction was induced, and after 72 h the infarction size was evaluated and the neurons and neuroglia in the perifocal zone were counted. APC reduced the infarction size 2.5 times in comparison with the control and reduced by 16% the neuronal death in the perifocal zone layer V, causing no appreciable changes in layer III, and did not change the size of neuronal bodies but increased (by 11%) the size of neuronal nuclei in layer III. The protein maintained the sharply increased count of gliocytes in the perifocal zone of infarction and promoted their growth. Hence, APC protected the neurons from death in the ischemic focus by increasing the gliocyte count and stimulating the compensatory reparative processes.

[EPR study of the influence of hypoxia on nitric oxide formation in the blood of Krushinskii-Molodkina rats].

The influence of hypoxia on nitric oxide formation in the blood of Krushinskii-Molodkina rats has been studied by electron paramagnetic resonance. It was found that nitric oxide synthesis in Krushinskii-Molodkina rats is increased compared with that in Wistar rats. A significant enhancement of the EPR signal of Hb-NO complexes in the animal blood was observed after hypoxia simulating the altitude of 5000 m above the sea level, in particular in the presence of sodium nitrite and the NO-synthase inhibitor L-nitroarginine. It was assumed that NO synthesis and nitro-/nitrite- reductase systems are activated under hypoxic conditions.

[Effect of ischemic preconditioning in ischemic insult by estimating the damage to low-density lipoproteins of rat blood by spectroscopic methods].

It has been shown by optical and spectral methods that ischemic preconditioning in ischemic insult induces a protective action on brain tissues, which is registered by an decrease in the damage to low-density lipoproteins after ischemic insult.

[Organ specificity in nitric oxide storage in the blood vessel walls during adaptation to hypoxia].

Addition of N-acetylcysteine induced relaxation of the coronary and basilar arteries thus indicating some basilar NO-stores in these vessels. The maximum capacity of the NO-stores was similar in the coronary and the basilar arteries. Following adaptation to hypoxia, however, the depot was much greater in the coronary artery wall. This seems to be connected with different degree of participation of the NO-dependent vasodiatation in implementation of the adaptive response to hypoxia in coronary and cerebral vascular systems.

[Nitric oxide is involved in the protective effects of short-term adaptaion to hypoxia in the course of stress-induced disorders in Krushinsky-Molodkina rats].

A possible involvement of nitric oxide in the protective effect of short-term adaptation of Krushinsky-Molodkina rats to mild hypoxia simulating 5000 m above sea level was studied. Nitric oxide proved to have a considerable protective effect on stress-induced disorders in Krushinsky-Molodkina rats as demonstrated using NO-synthase inhibitors and NO monitoring by electron spin resonance under different experimental conditions.

[Reduction of erythrocyte deformability in Krushinskii-Molodkina rats in acute hemorrhagic cerebrovascular circulation disorders].

The ability of erythrocytes to change their shapes in the shear flow under acute strokes of hemorrhagic type in rats of the Krushinsky-Molodkina line was studied. The rigidity of membranes and the internal viscosity of erythrocytes were investigated by the laser diffraction method. The method consists in obtaining diffraction images from a thin layer of a dilute suspension of erythrocytes moving in the shear flow and subsequent computer processing of these images. It was shown that strokes of hemorrhagical type in rats of the Krushinsky-Molodkina line cause a reduction in the ability of erythrocytes to change theirs shapes.

[Elevation of the vascular smooth muscle sensitivity to effects of constrictors after denervation and under decreased blood pressure].

Changes in contractile activity of saphenous artery in normotensive rats and in rats with regional hypotension have been investigated. The abdominal aorta was partially occluded in Wistar rats distally to the renal arteries. Four weeks later, a 5-7-mm segment of the femoral nerve in one hindlimb was resected to denervate the saphenous artery. After two weeks, the isometric contraction of innervated and denervated saphenous artery segments was studied. In normotensive rats, the denervation augmented vessel sensitivity to noradrenaline, phenylephrine, serotonin, and KCl (in the presence of phentolamine). Chronic hypotension also augmented vessel sensitivity to constrictor agonists, whereas denervation did not result in further increase of sensitivity. In glyoxilic acid-stained preparations obtained from hypotensive rats, a reduced intensity of fluorescence of adrenergic fibers was observed. It was assumed that the higher sensitivity of vascular smooth muscle in hypotensive rats is due to functional disturbances of sympathetic innervation.

[Neuroprotective activity of the proline-containing dipeptide noopept on the model of brain ischemia induced by the middle cerebral artery occlusion].

The influence of noopept (N-phenylacetyl-L-prolylglycine ethyl ester, GVS-111) on the extent of ischemic cortical stroke was investigated in experiments on white mongrel male rats with ischemia induced by a combination of the middle cerebral artery occlusion with ipsilateral common carotid artery ligation. Animals were treated with noopept (0.5 mg/kg, i.p.) according to the following schedule: 15 min and 2, 24, and 48 h after the occlusion. Test rats were decapitated 72 h after occlusion, brains were extracted and frozen, and thin brain slices were stained with 2,3,5-triphenyltetrazolium chloride. The slices were scanned and processed using Auc 1 computer program, which estimates the percentage of damaged area relative to that of the whole ipsilateral hemisphere. The conditions of coagulation the distal segment of middle cerebral artery were selected, which caused necrosis localized in the fronto-parietal and dorso-lateral regions of the brain cortex without any damage of subcortical structures. The extent of the brain damage in control group (treated by saline) was 18.6%, while that in the group treated with noopept was 12.2%, thus demonstrating a decrease in the infarction area by 34.5% (p < 05). The data on noopept efficacy on the model of the extensive ischemic injury of brain cortex show that this drug has good prospects for use in the neuroprotective treatment of stroke.

[Protective effect of peptide semax the rat heart in acute myocardial infarction].

Semax, a member of ACTH-derived peptides family, has been employed in the treatment of acute ischemic stroke in patients. It decreased neurological deficit and reduced NO hyperproduction in the rat brain, caused by acute cerebral hypoperfusion. We suggested that semax is also able to protect rat heart from ischemic damage in acute myocardial infaction (AMI). AMI was induced by left coronary artery occlusion, myocardial ischemic area averaged 30 % of left ventricle. In 2 hours after coronary occlusion, the AMI group developed 11 % reduced mean arterial blood pressure and 48 % increased diastolic blood pressure in left ventricle in comparison with sham-operated control group. However, infusion of either dobutamine, which directly stimulates myocardial contractility, or sodium nitroprusside and phenylephrine, that change vascular resistance and thus cardiac afterload, did not reveal distinctions in hemodynamic parameters between groups. These data indicate absense or only moderate cardiac dysfunction in rats with AMI and are consistent wih morphometrical and histochemical studies that did not detect any necrotic or apoptotic (TUNEL-test) changes in left ventricular cardiomyocytes in spite of development of distinct ischemic disturbances of mitochondria and nuclear in about 50 % of cardiomyocytes in 2 hours after AMI. Semax (150 microg/kg), given i. p. 15 min and 2 hours after coronary occlusion, caused no effect on cardiac function, but completely prevented ischemia-induced ultrastructural changes of cardiomyocytes. This protective effect was accompanied by the ability of peptide to blunt the increase in plasma concentrations of nitrates, observed in AMI group.

[Protective effect of peptide semax (ACTH(4-7)Pro-Gly-Pro) on the rat heart rate after myocardial infarction].

Semax, a member of ACTH-derived peptides family, was used in treatment of ischemic stroke in patients. It decreased neurological deficiency and reduced NO hyperproduction in the rat brain caused by acute cerebral hypoperfusion. We suggest that semax is also capable of protecting the rat heart from ischemic damage 28 days after myocardial infarction (MI) induced by left descendent coronary artery occlusion. Semax (150 microg/kg) was given i. p. in the operating day twice: 15 min and 2 hours after coronary occlusion, and once a day for the following 6 days. In 28 days after infarction, the MI group developed cardiac hypertrophy, cell growth was caused mainly by the increase of contractile filaments not supported by the appropriate mitochondrial growth that indicated an impaired energy supply of the cells. Moreover, cardiac hypertrophy was accompanied by decreased mean arterial blood pressure and cardiac contractile function and increased left ventricular end-diastolic pressure. Pharmacological change of cardiac afterload revealed that, in 28 days after MI, the rat heart was not able to change its contractile performance in response to either increase or decrease of systemic blood pressure, and as a result could not maintain its diastolic pressure. All these changes obviously reflect development of heart failure. Semax did not affect cardiac work but partially prevented end-diastolic pressure growth in left ventricle as well as ameliorated cardiomyocyte hypertrophy and disproportionate growth of contractile and mitochondrial apparatus, thus exerting beneficial effect on the left ventricular remodeling and heart failure development late after myocardial infarction.

Acute hypoxia during organogenesis affects cardiac autonomic balance in pregnant rats.

Changes in ECG parameters were studied in pregnant rats exposed to acute hypobaric hypoxia during the period of organogenesis (gestation days 9 to 10). Rats with low, medium, and high tolerance to hypoxia exhibited pronounced autonomic nervous system imbalance, which become apparent as a loss of correlation between various parameters of ECG signals recorded at rest and during exposure to some stress factors existing under normal conditions.

Reactivity of rat basilar artery to serotonin after short-term ischemia of hindbrain and during chronic vertebrobasilar insufficiency.

Contractile responses of the basilar artery to serotonin were examined in vitro on two models of circulation disturbances in the vertebrobasilar region of the brain. Two days after 30-min reversible occlusion of vertebral arteries, the sensitivity of the basilar artery to serotonin decreased, while chronic vertebrobasilar insufficiency had no effect on serotonin-induced contraction.

C-terminal Pro-Gly-Pro tripeptide in contrast to full-length neuropeptide semax exhibits no neuroprotective effect in experimental cerebral ischemia.

The C-terminal fragment Pro-Gly-Pro of semax does not modulate the development of symptoms of neurological deficiency and mortality in rats with incomplete global cerebral ischemia. Hence, previously revealed neuroprotective effects of semax are mainly determined by corticotropin ACTH4-7 fragment.

[Antistress and angioprotective influence of nitric oxide in epilepsy-prone rats of Krushinskii-Molodkina strain].

In rats of Krushinsky-Molodkina strain (KMR), the audiogenic stress induced epileptiform seizure and development of acute disturbances of cerebral circulation of hemorrhagic nature. The inhibitor of NO-synthase (L-NNA) increased the severity of clinical symptoms, mortality, and the intensity of intracranial hemorrhages. In contrast, L-arginine elevated the resistance of KMRs to acoustic stress. The intensity of nitrergic innervation was analyzed in preparations of the middle cerebral artery with the use of histochemical NADPH-diaphorase staining. In preparations of control KMRs, a net of NADPH-positive perivascular nerve fibers was found, while in experimental KMRs, in an hour after sound stimulation, such fibers practically were not revealed. Preliminary exposure of KMRs in hypoxic conditions (1 hour in hypobaric chamber at simulated altitude of 5000 m above the sea level) decreased the development of stress lesions. The protective effect of hypoxic training disappeared after the administration of NO-synthase inhibitor (L-NNA). The study demonstrated participation of nitric oxide (NO) in adaptive reactions of cerebral hemodynamics linked with the significant increase of cerebral blood flow.