Effect of ß-amyloid peptide 42 on the dynamics of expression and formation of Аß40, IL -1ß, TNF α, IL -6, IL -10 by peripheral blood mononuclear cells in vitro and its correction by curcumin.

The toxic effect of Аß-oligomers accompanies chronic inflammation, with cytokines as main mediators. Therefore, the cytokine link of inflammation becomes a new target on the way to restrain amyloidosis. The aim of the study was the effect of aggregated Аß42 on the dynamics of expression and formation of endogenous Аß40 and cytokines (IL-1ß, TNFα, IL-6, IL-10) by peripheral blood mononuclear cells in vitro and its correction by curcumin. A suspension of mononuclear cells isolated ex tempore using ficoll-urografin gradient from venous blood samples of healthy volunteers were used to study the effects of Аß42 (15 nM), curcumin (54 pM), and their combined action (at similar concentrations) in time dynamics: 0, 1, 3, 6 and 24 h incubation at 37 °C. Polymerase chain reaction with appropriate primers was used to determine the relative expression of mRNA for AßPP, TNFα, IL-1ß, IL-6, IL-10 and enzyme-linked immunosorbent assay ­ to determine the content of Аß40 and cytokines in mononuclear suspension during all periods of incubation. The individual dynamics AßPP and cytokine expression was shown under the action of the Aß42, which had influence on the content of Aß40, TNFα, IL-1ß, IL-6 and IL-10 in mononuclear suspension. Curcumin displayed the inhibitory effect on gene expression of AßPP, TNFα and IL6, which resulted in the decrease of the level of these two cytokines and Aß40. Thus, the dynamics of anti-inflammatory effect of curcumin in vitro for transcriptional and translational levels of cytokine's formation by mononuclear cells was shown in the work. Direct inhibitory effect of curcumin on the concentration of endogenous Aß40 during the 24 h incubation in conditions of toxic action of Aß42 aggregates was established.

Effect of curcumin on accumulation in mononuclear cells and secretion in incubation medium of Аß(40) and cytokines under local excess of Аß(42)-homoaggregates.

The aim of the work was to investigate accumulation of endogenous Aß40 and cytokines (IL-1ß, TNFα, IL-6, IL-10) in mononuclear cells and their secretion into incubation medium under Aß42-aggregates' toxicity and anti-inflammatory effects of curcumin. Mononuclear cells were isolated in Ficoll-Urografin density gradient from venous blood of healthy donors, resuspended and used for testing of homoaggregates of Aß42 (15 nM), curcumin (54 pM) and their combinations on various timescales (0, 1, 2, 3, 6 and 24 hours). Endogenous Aß40 and cytokines were detected in mononuclear cells and (separately) in incubation medium by ELISA. We demonstrated for the first time that homoaggregates of Aß42 cause rapid accumulation of endogenous Aß40 in mononuclear cells and accelerate its secretion into incubation medium. We found increased concentration of TNFα after 3 hours of incubation, and no changes in IL-1ß concentration due to secretion of these pro-inflammatory factors into incubation medium. The concentrations of IL-6 in mononuclear cells were increased under effects of Aß42 homoaggregates, and it was being secreted profoundly into incubation medium. Aß42 did not affect IL-10 secretion, yet caused an increase in its intracellular concentration after 1 hour of incubation, which was subsequently suppressed. Curcumin prevented the increase in Aß40 concentration in mononuclear cells and significantly decreased its secretion resulting from Aß42 toxicity. Curcumin negated the activating effect of Aß42 on pro-inflammatory cytokines, starting immediately for IL-1ß and on 3-6 hours for TNFα, which resulted in decreased extracellular concentrations of these cytokines. The polyphenol also potentiated repleni­shing of intracellular IL-6 and IL-10 concentrations and their secretion into incubation medium.

[Cytokines neuroinflammatory reaction to the action of homoaggregatic and liposomal forms of b-amyloid 1-40 in rats].

An injection model of preclinical stages of Alzheimer's disease has been reproduced in rats. It was accompanied by the decrease in the latent period of conditioned reflex avoidance, increasing levels of endogenous b-amyloid peptide 1-40 and activation of inflammatory cytokines (IL-1b, TNF-a, IL-6, IL-10) in the cerebral cortex, hippocampus and blood serum of experimental animals. We belive that changes identified at the biochemical level are prerequisite to modulate neuronal functions in rats induced by Ab40_Human administration. The toxic effect of exogenous b-amyloid 1-40 homoaggregates caused intense response of the cytokine system, while its liposomal form caused the soft information signal to the activation of innate immunity.

[Effect of valproic acid on SMN protein level in peripheral blood mononuclear cells of patients with spinal muscular atrophy and different SMN2 copy numbers].

OBJECTIVE: Spinal muscular atrophy (SMA) is currently untreatable hereditary disorder caused by few types of mutations in the SMN1 gene and respective lack of gene's product - survival motor neuron protein (SMN). Last decade studies have shown that phenotype of the disorder is substantially influenced by copy numbers of homologous SMN2 gene; also, an ability of valproic acid to increase the level of SMN in vitro and in vivo has been shown. We investigated an effect of valproic acid on SMN level in peripheral blood mononuclear cells derived from patients with SMA and their parents and sought for possible predictors for treatment efficacy. MATERIAL AND METHODS: We examined 10 children with SMA and 6 their parents as heterozygous carriers of the mutation using appropriate molecular-genetic techniques. Valproic acid was prescribed in 20mg/kg/day during 2 weeks. RESULTS AND CONCLUSION: There were no correlation between baseline SMN level and SMN2 copy number; both of the markers do not predict SMN level after the treatment with valproic acid. However, all of patients responded to valproic acid treatment with different grades of SMN level increase. Strong intrafamilial correlation was found for the SMN/Β2-microglobulin ratio.

[Intensive protein synthesis in neurons and phosphorylation of beta-amyloid precursor protein and tau-protein are triggering factors of neuronal amyloidosis and Alzheimer's disease].

Recently the studies of Alzheimer's disease have become particularly actual and have attracted scientists from all over the world to this problem as a result of dissemination of this dangerous disorder. The reason for such pathogenesis is not known, but the final image, for the first time obtained on microscopic brain sections from patients with this disease more than a hundred years ago, is well known to clinicists. This is the deposition of Abeta amyloid in the brain tissue of senile plaques and fibrils. Many authors suppose that the deposition of beta-amyloid provokes secondary neuronal changes which are the reason of neuron death. Other authors associate the death of neurons with hyperphosphorylation oftau-proteins which form neurofibrillar coils inside nerve cells and lead to their death. For creation of methods of preclinical diagnostics and effective treatment of Alzheimer's disease novel knowledge is required on the nature of triggering factors of sporadic isoforms of Alzheimer's disease, on cause-effect relationships of phosphorylation of amyloid precursor protein with formation of pathogenic beta-amyloids, on the relationship with these factors of hyperphosphorylation of tau-protein and neuron death. In this review we analyze the papers describing the increasing of intensity of biosynthesis in neurons in normal conditions and under the stress, the possibility of development of energetic unbalanced neurons and activation of their protective systems. Phosphorylation and hyperphosphorylation of tau-proteins is also tightly connected with protective mechanisms of cells and with processes of evacuation of phosphates, adenosine mono-phosphates and pyrophosphates from the region of protein synthesis. Upon long and high intensity of protein synthesis the protective mechanisms are overloaded and the complementarity of metabolitic processes is disturbed. This results in dysfunction of neurons, transport collapse, and neuron death.

[Characteristics of lipid composition of blood serum lipoproteins in insulin resistance].

The purpose of this investigation was to study lipid composition of apoB- and apoA-containing lipoproteines in conditions of insulin resistance pathology. It was shown that hypertriglycerolemia and hypercholesterolemia were determined by triglycerole and cholesterol surplus in the composition of both apoB- and apoA-containing lipoproteins. Phospholipid deficiency in the composition of all lipoproteins fractions were established. Possible mechanisms of formation and metabolism in circulation of blood lipoproteins with altered lipid composition in the blood flow at insulin resistance is discussed.

[The lipoprotein metabolism in arterial hypertension, type II diabetes mellitus, and metabolic syndrome].

The apoA- and apoB-containing lipoprotein (LP) fractions, activity of free and linked forms of lipoprotein lipases (LPL) of liver and extrahepatic tissues, and also activity of lecithin:cholesterol acyltransferase (LCAT) were examined in the blood serum of patients with arterial hypertension, type II diabetes mellitus (DM-II), and metabolic syndrome (MS). Patients with DM-II had the most pronounced changes of all investigated LP fractions compared with healthy persons and patients of other groups examined. Decrease of LCAT activity corresponded to declining level of apoA-containing LP in DM-II and MS. Based on these data obtained we suppose, that one of peculiarities of LP metabolic disorders in the patients is a releasing of a part of lipolitic enzymes, located on the capillary endothelium, into blood flow. Heparin may have an important role in LPL redistribution, as its concentration in blood cells was declined in all the patients.

[Depression of serum esterase and lipoprotein lipase activities in acute and longitudinal actions of ethanol].

Esterase (LCAT) and lipoprotein lipase (LPL) activities of blood serum, and a range of blood serum lipoproteins (LP) in acute and chronic ethanol intoxication were examined in healthy persons and patients with alcohol abuse. Ethanol inhibited LCAT and LPL activities, and increased apoA-containing LP in blood serum. These changes are considered as a basis for depression of a direct and reverse transportation of cholesterol in blood circulation under the action of ethanol.

[Changes in the lipoprotein metabolism in alcoholism do not recover during of prolonged remission].

Contents of lipoproteins (LP), activity of lipoprotein lipase (LP-lipase) and cholesterol esterification activity of blood serum were examined in patients with alcoholism at the periods of intoxication, alcohol abstinence and under conditions of prolonged remission. During remission a general pattern of apoB-containing LP was similar to the period of intoxication. Processes of transformation of this class LP significantly delayed in isolated blood serum in vitro. The magnitude of these changes almost corresponded to the period of intoxication. However activity of free and membrane-bound forms of LP-lipase. A high level of LPHD2a was characteristic for the conditions of intoxication and alcohol abstinence. In the remission the level of all the apoA-containing LP fractions was significantly lower than in the control. Besides, processes of their transformations in vitro were slowed down. Intensity of these changes resembled the period of intoxication. Thus, disorders of LP metabolism pointed out during the period of intoxication and alcohol abstinence do not recovered under conditions of a prolonged remission.

[A short-term effect of lovastatin on lipoprotein metabolism in patients with cerebrovascular lesions concomitant with diabetes mellitus II type].

Changes of lipoprotein (LP) metabolism after lovastatin therapy (20 mg/day for 2 months) have been studied in patients with cerebrovascular lesions (CBL) concomitant with diabetes mellitus type 2 (DM-2) and in CBL patients without DM-2. Initial effect of lovastatin appeared in decreasing chilomicrone content in both groups of patients. A distinct trend towards restoration of processes converting apoA-containing LP being accompanied by normalization of function of the peripheral lipoprotein lipase bound form was observed. The data obtained support an opinion about rationality of lovastatin treatment of dyslipidemias in CBL concomitant with DM-2 as well as of cardiovascular pathology.

[Lipoprotein metabolism in patients with diabetes of type II].

Changes in contents of blood serum lipoproteins (LP) and activity of enzymes of their transformation were investigated in patients with diabetes mellitus of type II. It was found out that mechanisms of increasing of apoB-containing LP in these patients were on the one hand in enhancing of the processes of their new-formation in the liver. On the other hand, retardation of their arrival to peripheral tissues might be observed, that as a whole resulted in continuous LPLD circulation in blood. Above-mentioned processes were associated with a reciprocal decreasing of the amount of all the apoA-containing LP fractions examined. To a certain extent mechanisms of disturbance of LP metabolism in the circulation bed were stipulated by changes of function of the enzyme catalyzing their transformation.

[Role of heparin in the formation of food lipemia].

Influence of endogenic and exogenic heparin in vivo on the basic forms of serum lipids content: cholesterol ethers, triacylglycerols, free fatty acids; as well as that glycosaminoglycan effect in vivo and in vitro on total lipoproteine lipase (LPL) activity and lecithin-cholesterol acyltransferase (LCAT) activity of human blood serum were investigated on food lipidemia model. The decrease of intercell reserve heparin content and increase of the background and post-heparin levels of blood serum LPL activity were indicated after two hours food load. The role of two factors, endogenic heparin being one of them, in the increase of postprandial LPL activity of blood serum were discussed. At the same time, some inhibition of blood serum LCAT activity two hours after food reception (evidently, as a result of endogenic heparin action) and to a considerable extent inhibition of cholesterol etherification under the action of exogenic heparin in vivo were ascertain. Heparin in vitro (50 U/ml of blood serum) did not influence LCAT and total LPL activities. It was summarised that endogenic heparin is a factor, taking part in lipolysis processes regulation.

[Lipid-mobilizing effect of reduced glutathione and its intensifying action on lecithin-cholesterol acyltransferase activity in blood serum of rats]. / Lipidmobilizuiushchii éffekt vosstanovlennogo glutationa i ego potentsiruiushchee deistvie na letsitin-kholesterol atsiltransferaznuiu aktivnost' v syvorotke krovi krys.

Effect of reduced glutathione (50 mg/100 g) on lipid distribution between organs (liver and kidney) and lecithin-cholesterol acyltransferase (LCAT) activity in blood serum of rats was investigated. The accumulation of common lipids as a result of speeding up the absorbtion of blood serum unsaturated fatty acids and relative decrease of lipids unsaturation in the liver and lipid content dynamics in kidneys owing to the intensification of two processes in this organ: the transport of polyene fatty acids in composition of blood serum lipoprotein lipids to kidney cells and peroxidation of membrane phospholipids were found out. The activating effect of GSH (in vivo and in vitro) on LCAT activity of rat blood serum was shown. It was summarised that GSH-intensification of blood serum etherification ability may be a basic component of reduced glutathione lipid mobilization effect.

[Investigation of lipoprotein metabolism in persons with abstinence]. / Issledovanie obmena lipoproteinov u abstinentov.

Lipoprotein (LP) metabolism was investigated in the blood serum of persons with abstinence after ethanol intake in a single dose of (0.7-1.0) g/kg. Content of LP was determined with the method of gradient gel-electrophoresis and use of software for quantitative analysis. A LP-lipase and cholesteroletherificazing activity of blood serum were also examined. It was found out that the patients with abstinence were characterized by the slowing down of LP metabolism as compared with donors, who represented general population. A single action of ethanol causes a short-time inhibition of LP metabolism. After 3 days processes of apoA-containing LP transformation were renewed. In contrast to this, one could observe an increase of the level of apoB-containing fractions accompanied by incompatibility between quantitative LP changes and activities of enzymes, which catalyzed their transformation. This evidences for the impairment of coordination of processes of cholesterol and triacylglyceroles transport by ethanol.

[Content and composition of lipoproteins of rat blood and liver and various parameters of oxidative stress during administration of cobalt chloride]. / Soderzhanie i sostav lipoproteinov krovi i pecheni krys i nekotorye pokazateli okislitel'nogo stressa pri vvedenii khlorida kobal'ta.

Cobalt chloride effect on rat liver and serum blood lipoproteins content and composition and on some characteristics of lipid peroxidation and oxidative stress was investigated. The activation of free-radical oxidation and oxidative stress development were judged from the dynamics of lipid peroxidation products accumulation, from cathepsin D unsedimental activity and from the alteration of microsomal cytochrome P-450 content and from activity of a number antioxidative enzymes. In order to evaluate the state of glutathione-defence system the activities of glutathione peroxidase, glutathione S-transferase, glutathione reductase and some NADPH-generating enzymes and reduced glutathione level alteration were studied in liver. The data obtained show that the cobalt chloride injection leads to the development of the oxidative stress and to activation of some antioxidant defence system, namely, glutathione-depending enzymes, and of microsomal cytochrome P-450 catabolism. The system blood lipoproteins (liver lipoproteins was found to participate in metabolism adaptation under oxidative stress and in maintenance of biological membranes structure and functioning.