[Influence of the rat diet enrichment with oat ß-gucans on the assimilation of B group vitamins, mineral elements and lipid metabolism].

Despite the widespread use of oat ß-glucans as ingredient of foods and dietary supplements, there is insufficient data on their effect on the metabolism of vitamins and minerals. The purpose of the study was to evaluate the effect of including oat bran with a high content of ß-glucans (ß-glucan) in the diet on the absorption of micronutrients and lipid metabolism in growing rats deficient in vitamins D, group B and trace elements (iron, copper, zinc). Material and methods. After the development of micronutrient deficiency (for 23 days), in order to assess the effect of oat bran (5%) with a high content of ß-glucans on the correction of the micronutrient status of growing male Wistar rats (with initial body weight of 70.7±0.7 g), the missing micronutrients were introduced in the semi-synthetic diet deficient in vitamins D, group B, iron, copper and zinc within 7 days either along with ß-glucan (1.47%) or without its addition. Indicators of micronutrient sufficiency (riboflavin serum concentration, daily urinary excretion of thiamine, riboflavin and 4-pyridoxic acid, measured by fluorometric methods; serum concentration and urinary excretion of calcium, magnesium, iron, zinc, copper, phosphorus, measured by the atomic absorption method or using standard methods on a biochemical analyzer) and the biochemical parameters of blood serum were compared with the parameters of rats adequately provided with all micronutrients throughout the experiment. Results. Replenishment of missing micronutrients in the diet of rats with deficiency in vitamins D and group B, iron, copper and zinc for 7 days led to the elimination of deficiency of vitamins B1, B2 and B6, regardless of the presence of ß-glucans in the diet. At the same time, against the background of the presence of ß-glucans in the feed, an increase in the absorption of iron was observed, as evidenced by an increase by 1.73 times in iron blood plasma level (р<0.05) and a tendency towards its urinary excretion decrease by 1.60 fold (р<0.10) compared to animals from the control group. Adding oat bran with ß-glucans to the feed did not lead to a decrease in blood plasma level of total cholesterol and low-density lipoproteins cholesterol. The levels of high-density lipoprotein cholesterol and triglycerides in rats of all three groups did not have statistically significant differences. Conclusion. The presence of ß-glucans in the diet had virtually no effect on the absorption of B vitamins and improved the absorption of iron.

[Pathogenetic mechanisms for the development of hematological disorders in induced fatty liver disease in Wistar rats and assessment of the regulatory effects of carnosine and α-lipoic acid].

The study of the relationship between hematopoiesis and metabolism is now particularly relevant in view of the high incidence of alimentary dependent diseases, including non-alcoholic fatty liver disease. In this regard, pathogenetic factors of this disease development are studied actively in order to choose adequate drug therapy and usage of bioactive substances with antioxidant properties. The aim of the study was to study the pathogenetic relationship of hematological disorders and imbalance of growth factors, leptin and ghrelin in male Wistar rats in the model of the initial stage of non-alcoholic fatty liver disease development and to assess the regulatory effect of minor bioactive substances - carnosine and α-lipoic acid. Material and methods. The studies were performed on male Wistar rats with initial body weight 150±10 g within 8 weeks. Animals were divided into 5 groups (n=8 in each). Rats of the control group received a complete modified diet AIN93M, in which soybean oil was replaced with sunflower oil and lard (1:1). Rats of the experimental groups consumed high-calorie choline-deficient diet (HCCDD), in which fat content was 45%, fructose content - 20% of the energy value of the diet. Rats of the 2nd group were fed HCCDD without any supplements, the 3rd group - with the addition of carnosine (75 mg/kg body weight), the 4th group - with the addition of α-lipoic acid (75 mg/kg body weight), the 5th group - with the combined addition of carnosine and α-lipoic acid in a total dose of 150 mg/kg body weight. Hematological values were determined on a hematological analyzer. The content of ghrelin and leptin, as well as growth factors GM-CSF and M-CSF in blood plasma and adipose tissue lysates, was determined by multiplex immunoassay using xMAP technology. Results. Rat intake of HCCDD resulted in decreased hemoglobin levels and red blood cell scores compared to controls. Diet enrichment with carnosine and α-lipoic acid did not have a reliable effect on these indicators. Carnosine intake had a protective effect on erythrocyte volume, a decrease of which was recorded in other experimental groups. HCCDD stimulated the growth of the absolute number of leukocytes in peripheral blood due to granulocytes and mononuclears. The enrichment of HCCDD with carnosine and α-lipoic acid led to a further increase in leukocytosis, the maximum level of which was observed in the group of rats fed HCCDD, simultaneously enriched with carnosine and α-lipoic acid (14.86±1.48×109/l compared to 8.67±1.23×109/l in control). All diets used in the research had no effect on the number of erythrocytes and platelets in the peripheral blood of rats. The use of both HCCDD alone and in combination with carnosine or α-lipoic acid intake had a negative effect on the level of growth factors GM-CSF and M-CSF in blood plasma and adipose tissue. The consumption of HCCDD caused an increase in leptin blood level (8.54±0.69 compared to 2.58±0.37 pg/ml in control, р<0.05), which was normalized by enriching the diet with carnosine and α-lipoic acid. Ghrelin blood level significantly decreased in all experimental groups compared to the control: by 30% in rats fed and by almost 50% when carnosine and α-lipoic acid were added to HCCDD. The intake of α-lipoic acid led to hormone level changes in adipose tissue lysates, leptin content decreased (2.31±0.11 vs 2.77±0.15 pg/ml), while ghrelin level significantly increased (0.35±0.14 vs 0.20±0.06 pg/ml), compared with the control group (р<0.05). Conclusion. The revealed interrelation of parameters of the cellular composition of peripheral blood and hemoglobin content with the changes in the content of GM-CSF, M-CSF, leptin and ghrelin in blood plasma and adipose tissue indicates the mutual influence of the studied CSF, leptin, ghrelin and added antioxidants (carnosine and α-lipoic acid) on the regulatory mechanisms of hematopoiesis in rats fed HCCDD.

[Effect of carnosine and α-lipoic acid on hepatocyte apoptosis and the cytokine profile in induced fatty liver disease in Wistar rats].

Non-alcoholic fatty liver disease (NAFLD) is now a common liver disease affecting about a third of the world's population. In this regard, the issue of studying the pathogenetic factors of the development of this disease in order to select adequate drug therapy and biologically active substances with antioxidant properties regulating the balance of pro- and anti-inflammatory cytokines is of particular relevance. The aim of the study was to assess the effect of minor biologically active substances - carnosine and α-lipoic acid on hepatocyte apoptosis and the cytokine profile in the experimental model of the initial stage of NAFLD. Material and methods. The studies were performed on male Wistar rats with initial body weight of 150±10 g. Animals were divided into 5 groups of 8 rats each. Within 8 weeks, rats of the 1st group (control) received a complete modified diet AIN93M, in which soybean oil was replaced with sunflower oil and lard (1:1). Rats of the experimental groups consumed high-calorie choline-deficient diet (HCCDD), in which fat content was 45%, fructose content - 20% of the energy value of the diet. Rats of the 2nd group were fed HCCDD without any supplements, the 3rd group - with the addition of carnosine (75 mg/kg body weight), the 4th group - with the addition of α-lipoic acid (75 mg/kg body weight), the 5th group - with the addition of carnosine and α-lipoic acid in a total dose of 150 mg/kg body weight. The study of rat hepatocyte apoptosis was performed by flow cytometry. Hepatocytes were stained with annexin V and vital dye 7-aminoactinomycin, followed by detection on an flow cytometer. The content of cytokines and chemokines (IL-1α, IL-10, IL-17А, M-CSF, MIP-1α, MIP-3α, RANTES) in the cytoplasmic fraction of liver tissue was determined by multiplex immunoassay. Results and discussion. On the model of the initial stage of development of NAFLD in male Wistar rats the еnrichment of HCCDD with carnosine and α-lipoic acid had demonstrated a protective effect on hepatocytes with a decrease in apoptosis intensity to the level in control rats. Under the influence of HCCDD, an increase in the content of M-CSF and MIP-1α and a decrease in the levels of MIP-3α and RANTES, stimulating the migration and differentiation of various immunoregulatory populations to the parenchyma at an early stage of the formation of fatty hepatosis, in the cytoplasmic fraction of liver tissue were detected. Moreover, a decrease in the level of proinflammatory cytokines IL-17A and IL-1α and an increase in IL-10 produced mainly by Treg-populations indicate the absence of pronounced inflammatory changes in the liver of male Wistar rats at the initial stages of development of fatty dystrophy. Conclusion. Enrichment of HCCDD with both carnosine and α-lipoic acid in Wistar rats had a protective effect on hepatocytes with a decrease in apoptosis to a level in control rats. The increase in the IL-10/IL-17A ratio indicates the activation of anti-inflammatory mechanisms due to the functional predominance of Treg-cells over Th1/Th17 lymphocytes.

[Effects of rutin on protective capacity in rats].

The purpose of the study was to determine effects of rutin dietary administration on the activity of xenobiotic-metabolizing enzymes and antioxidant status. The study has been carried out on 3 groups of male Wistar rats (n = 8 in each), with initial body weight 100-120 g. Animals of the control group (1st group) received standard semi-synthetic diet, the experimental groups--the same diet with rutin in the amount of 40 mg/kg b.w. (2nd group) or 400 mg/kg b.w. (3rd group). The duration of the experiment was 2 weeks. In rat liver the activity of quinone reductase (QR), heme oxygenase-1 (HO-1), paraoxonase-1 (PON-1), glutathione-S-transferase (GST), ethoxyresorufin-O-dealkylase (EROD) activity of CYP1A1, methoxyresorufin-O-dealkylase (MROD) activity of CYP1A2, tes- tosterone 6ß-hydroxylase (6ß-TG) activity of CYP3A, total antioxidant activity (AOA) and malondialdehyde (MDA) content have been investigated. The expression of genes CYP1A1, CYP1A2 and CYP3A has been measured by reverse transcription-polymerase chain reaction (RT-PCR). The stability of lysosome membranes was estimated by the change of unsedimentable activity of lysosomal enzymes--arylsulfatase, ß-galactosidase and ß-glucuronidase. Rutin administration led to dose-dependent increase in the activity of antioxidant enzymes. In rats of the 3rd group received high-rutin diet the activity of QR, HO-1, PON-1 and GST increased by 68, 29, 17 and 22%, respectively, compared to the control (1st group); MDA level and AOA have not changed. Activity of EROD and MROD in liver microsomes of rats treated with rutin at a dose of 40 mg/kg b.w. (2nd group) increased by 33 and 58%, respectively, with a moderate increase in mRNA level of CYP1A1 and CYP1A2. Increasing the dose of rutin up to 400 mg/kg b.w. (3rd group) resulted in the decrease of the degree of EROD and MROD activation by 18 and 15%, respectively, compared to the 2nd group. Rutin had no significant effect on the activity of 6ß-TG and on the expression of CYP3A1 gene. Rutin dietary administration led to dose-dependent reduction of the unsedimentable activity of lysosomal enzymes, indicating the strengthening of the stability of lysosomal membranes. Thus, the obtained results showed that in healthy, intact rats high doses of rutin in the diet moderately but statistically significantly activate enzyme systems responsible for the protective and adaptive capacity of the organism.

Effect of polyunsaturated fatty acids ω-3 on the induction of activity and expression of CYP1A1 and CYP1A2 genes in the liver of rats under the influence of indole-3-carbinol.

Supplementation of the ration with eicosapentaenoic and docosahexaenoic ω-3 polyunsaturated fatty acids (PUFA) in doses of 0.3 and 1 g/kg body weight for 4 weeks had no effect on ethoxyresorufin O-dealkylase (EROD) activity and expression of the CYP1A1 gene in male Wistar rats, but caused a dose-dependent increase in methoxyresorufin O-dealkylase (MROD) activity of CYP1A2 (by 28 and 73%, respectively) without significant changes in CYP1A2 mRNA expression. ω-3 PUFA had no effect on the indole-3-carbinol-induced (20 mg/kg body weight over the last 7 days of the experiment) EROD activity and expression of CYP1A1 mRNA. The indole-3-carbinol-induced MROD activity was shown to increase by 6.2 times in rats not receiving ω-3 PUFA and only by 3.9 and 2.7 times in animals receiving ω-3 PUFA. The indole-3-carbinol-induced expression of CYP1A2 mRNA slightly increased in animals receiving ω-3 PUFA. Our results suggest that the effect of ω-3 PUFA on the induced and basal activity of CYP1A2 is not related to modulation of CYP1A2 gene expression.

[Effects of omega-3 polyunsaturated fatty acids on antioxidant capacity in rats].

Dietary administration of omega-3 polyunsaturated fatty acids (EPA + DHA, 1,2:1) at dose 0.3 or 1 g/kg bw during 4 weeks led to minor (by 14% and 17%, p<0.05) decrease of serum antioxidant capacity and serum level of vitamin E (by 30% and 31%, p<0.05) and the activity of paraoxonase-1 (by 14% at 0.3 g/kg bw, p<0.05). The activity of antioxidant enzymes in liver increased in a dose-dependent manner. At higher dose of omega-3 polyunsaturated fatty acids a 45% increase in the activity of paraoxonase-1 (p<0.05), 21% -heme oxygenase-1 (p>0.05), 68% - quinone reductase (p<0.05), 19% - glutathione S-transferase (p<0.05) compared to the control group was found. The direct relationship between activities of enzymes and increase of MDA level in liver (by 47 and 107%, p

[Effect of dietary fat on vitamin status of rats].

The influence of low (1%) and high (31%) diet fat content (sunflower-seed oil and lard 1:1 at a ratio of 1:1) on vitamin A, E, B1 and B2 status of growing Wistar rats (8 rats per group) with initial body weight 80-100 g has been investigated. The semi-synthetic diet contained vitamin mixture in doses covering the physiological requirement of these animals. The increase of fat content (31%) in the diet due to the presence of vitamin E in sunflower-seed oil automatically lead to 1,7-fold increase consumption of this vitamin compared to the control group. Diet fat content did not affect the level of vitamins B1 and B2 in rat liver. Excessive intake of fat and vitamin E for 6 weeks did not influence on the content of blood plasma vitamin E and rat liver vitamin A occurs at the same time, while significant 1,9 fold elevation of liver vitamin E level and 26 per cent increase of blood plasma vitamin A concentration. The almost complete exclusion of fat from the diet had no effect on blood plasma level of alpha-tocopherol and retinol, but resulted in a significant decrease of vitamins A and E content in rat liver by 40 per cent, indicating a deterioration of sufficiency with these fat-soluble vitamins. The analysis of the results obtained in this investigation and literature data have suggested that under excessive as well as under decreased consumption of fat there is a risk of the development of polyhypovitaminosis. Vitamin complex supplementation is required to prevent a possible worsening of vitamin status under diets with modified fatty component.

[Effects of dietary fat level on the xenobiotic metabolism enzymes activity and antioxidant enzymes in rats].

Male Wistar rats received fat-free diet or diets containing 5, 10 and 30% of fat (sunflower oil + lard, 1:1) for 4 weeks. The direct relationship between dietary fat level and ethoxyresorufin O-dealkylase activity of CYP1A1, methoxyresorufin O-dealkylase activity of CYP1A2, pentoxyresorufin O-dealkylase activity of CYP2B1 and testosterone 6beta-hydroxylase activity of CYP3A was found. Activities of key enzymes of phase II xenobiotic metabolism (total activity of glutathione transferase, activity of UDP-glucuronosyle transferase) and antioxidant enzymes (catalase, glutathione peroxidase, glutathione reductase, paraoxonase-1 and heme oxygenase-1) also increased with higher dietary fat level.

[Studies of effects of aluminum oxide nanoparticles after intragastric administration].

Growing Wistar rats received intragastrically nanoparticles (NPs) of aluminum oxide (Al2O3) daily during 28 days at doses of 1 or 100 mg per kg body mass. There were studied body mass of animals, relative mass of internals, rate of protein macromolecules absorption in the gut, oxidative damage of DNA, pool of tissue thiols, activity of hepatic enzymes of xenobiotic detoxication system, biochemical and hematological blood indices, stability of lysosome membranes, condition of antioxidant defense system, apoptosis of hepatocytes. Conducted experiments didn't reveal any marked toxic action of Al2O3 NPs on rats after 28 days of administration both in high and low dose. Among effects probably related to NPs influence on animals there were lowering of relative liver and lung masses, decrease of hepatic thiol pool, activity of CYP1A1 isoform in liver and glutathione reductase in erythrocytes, increase of diene conjugates of fatty acids in blood plasma. Said shifts were small in magnitude, didn't come out of margins of physiological norm and didn't show any distinct relation to NPs dose. However considering great importance of this nanomaterial as probable environmental contaminant the studies of it's toxicity must be continued in conditions of low doses (less than 1 mg per kg body mass) for long period of time.

[Effects of green tea extract and its components on antioxidant status and activities of xenobiotic metabolizing enzymes of rats].

Dietary administration of green tea extract (GTE) or epigallocatechin gallate (EGCG), quercetin (Qu) or caffeine (Cf) in doses equal to their concentration in GTE led to an increase of serum and liver antioxidant capacity and strengthening stability of microsomal and lysosomal membranes in rats. The antioxidant efficiency of EGCG and Qu was considerably higher than that of GTE. There were significant differences in the effects of EGCG, Qu and GTE on the activities and expression of mRNA for CYP1A1, CYP1A2 and CYP3A1. But feeding both GTE and Cf to rats results in similar elevated activities of CYP1A1, CYP1A2, UDP-glucuronosyl transferase and glutathion transferase. Our results suggest that Cf is the main contributor to GTE effects on activities of xenobiotic metabolizing enzymes.

[The study of apoptosis of hepatocytes of rats for estimation safety and efficacy of application biologically active substances].

The magnitude of apoptosis of hepatocytes of rats that were within 2 weeks on a balanced complete semi-synthetic diet supplemented with extract of green tea, epigallocatechin gallate (EGCG), quercetin, coffein (1-4th experimental groups accordingly) is studied. The contents EGCG, quercetin and coffein in rats of 2-4th experimental groups correspond to their content in diet of rats 1 experimental group, and equally contents in tonic beverage. It is determined that application in diets extract of green tea and its compounds does not damage effect on hepatocytes of rats. The use of diets with the inclusion of extract of green tea, quercetin and coffein provides the clear tendency to stabilize the plasma membrane of hepatocytes comparison with rats of control group (rats fed the same diet but without the inclusion of the above-mentioned biologically active substances).

Effects of combined treatment with resveratrol and indole-3-carbinol.

Male Wistar rats received a semisynthetic diet with resveratrol (100 mg/kg), indole-3-carbinol (20 mg/kg), or a mixture of these compounds in the same doses for 1 week. Activities of ethoxyresorufin dealkylase (EROD), methoxyresorufin dealkylase (MROD), pentoxyresorufin dealkylase (PROD), and 6ß-testosterone hydroxylase (6ß-TH) and the content of mRNA for CYP1A1, CYP1A2, and CYP3A1 were elevated in the liver of rats receiving indole-3-carbinol. These changes were accompanied by an increase in activity of phase II xenobiotic metabolism enzymes (quinone reductase, hemoxygenase-1, glutathione transferase, and UDP glucuronosyl transferase). Resveratrol did not modify activity of these enzymes. After combined treatment with the test compounds, resveratrol suppressed the indole-3-carbinol-induced increase in activities of EROD, MROD, PROD, and 6ß-TH, and expression of the corresponding genes. Combined treatment was characterized by potentiation of the antioxidant effects of these compounds.

[Effect of soybean isoflavones on antioxidant status in rats fed diet with oxidized linseed oil]. / Vliianie izoflavonov soi na antioksidantnyi status krys, poluchavshikh ratsion s okislennym l'nianym maslom.

The maintenance of male Wistar rats on semi-synthetic diets containing oxidized flaxseed oil for 4 weeks caused decrease in serum total antioxidant activity, significant suppression of microsomal UDP-glucuronosyl transferase activity and benzpyrene hydroxylase activity, increase in reduced glutathione level and catalase activity of liver. Supplementation of diets with 0.05% of soy isoflavones has resulted in normalization of investigated characteristics.

[Medical-biological evaluation of genetically-modified sugar beet line 77 (chemical composition and toxicologico-biochemical studies)]. / Mediko-biologicheskia otsenka geneticheski modifitsirovannoi sakharnoi svekly linii 77 (khimicheskii sostav i toksikologo-biokhimicheskie issledovaniia).

The rats were fed with the Genetically Modified Sugar Beet line 77 (Monsanto Ko, USA) 10 g/rat/day for 1 month. Their blood, urea and liver were investigated to measure total protein and glucose levels, aminotransferase and alkaline phosphatase activities, pH, creatinine level as well as hepatic enzyme activity of the I and II phases of xenobiotic metabolism and whole and non-sedimentated lysosomal enzyme activities and activity of antioxidant system.

[Medico-genetic evaluation of sugar sand, obtained from genetically-modified sugar beet line 77 (toxicologico-biochemical studies)]. / Mediko-biologicheskaia otsenka sakhara-peska, poluchennogo iz geneticheski modifitsirovannoi sakharnoi svekly linii 77 (toksikologo-biokhimicheskie issledovaniia).

The rats were fed with the Suger from the Genetically Modified Suger Beet line 77 (Monsanto Ko, USA) 3 g/rat/day for 3 months. Their blood, urea and liver were investigated to measure total protein and glucose levels, aminotransferase and alkaline phosphatase activities, pH, creatinine level as well as hepatic enzyme activity of the I and II phases of henobiotic metabolism and whole and non-sedimentated lysosomal enzyme activities and activity of antioxidant system.