CCl4-induced Hepatitis Research Articles

Induction of MTHFD2 in Macrophages Inhibits Reactive Oxygen Species-mediated NF-κB Activation and Protects against Inflammatory Responses.

The one-carbon metabolism enzyme methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) is critical for cancer cell proliferation and immune cell phenotypes, but whether it can contribute to macrophage inflammatory responses remains unclear. In this study, we show that MTHFD2 was upregulated by LPS in murine macrophages upon activation of the TLR4-MyD88-IKKα/β-NF-κB signaling pathway. MTHFD2 significantly attenuated LPS-induced macrophage proinflammatory cytokine production through its enzymatic activity. Notably, ablation of myeloid MTHFD2 rendered mice more sensitive to septic shock and CCl4-induced acute hepatitis. Mechanistically, MTHFD2 restrained IKKα/β-NF-κB activation and macrophage inflammatory phenotype by scavenging reactive oxygen species through the generation of NADPH. Our study reveals MTHFD2 as a "self-control" mechanism in macrophage-mediated inflammatory responses.

Hepatoprotective effect of 2,6-dimethylpyridine N-oxide (Ivin) in experimental model of CCl(4)-induced hepatitis of rats

The effect of concomitant pesticides and plant growth regulators on humans is still not well understood. N-oxide-2,6-dimethylpyridine (Ivin) is the plant growth regulator known to reduce the acute toxicity of pesticides, but its protective mechanisms need to be investigated. The aim of the study was to assess the hepatoprotective ability of 2,6-dimethylpyridine N-oxide (Ivin) using a model of acute CCl4-induced hepatitis. Male Wistar Han rats received two subcutaneous CCl4 injections (0.8 ml/100 g b.w.). Oral Ivin (13 or 0.13 mg/kg) and hepatoprotector “Silybor-35” (5 mg/kg) as reference substances were administered orally one hour pre- and 2 hours post-CCl4 injection. The biochemical assay of blood plasma, estimation of lipid peroxidation products in the liver tissue and histological liver analysis were done. The results of functional tests and histomorphological studies of liver tissue demonstrated that Ivin exhibited a pronounced hepatoprotective effect, more pronounced when it was administered at a low 0.13 mg/kg dose. Calculation of the hepatoprotection efficiency index for Ivin showed that it was comparable to that for “Silybor-35”. Keywords: hepatoprotective effect, Ivin, СCl4-induced hepatitis of rats

Bioluminescence imaging of mouse monocyte chemoattractant protein-1 expression in inflammatory processes.

Monocyte chemoattractant protein-1 (MCP-1) plays a crucial role in various inflammatory diseases. To reveal the impact of MCP-1 during diseases and to develop anti-inflammatory agents, we establish a transgenic mouse line. The firefly luciferase gene is incorporated into the mouse genome and driven by the endogenous MCP-1 promoter. A bioluminescence photographing system is applied to monitor luciferase levels in live mice during inflammation, including lipopolysaccharide-induced sepsis, concanavalin A-induced T cell-dependent liver injury, CCl 4-induced acute hepatitis, and liver fibrosis. The results demonstrate that the luciferase signal induced in inflammatory processes is correlated with endogenous MCP-1 expression in mice. Furthermore, the expressions of MCP-1 and the luciferase gene are dramatically inhibited by administration of the anti-inflammatory drug dexamethasone in a septicemia model. Our results suggest that the transgenic MCP-1-Luc mouse is a useful model to study MCP-1 expression in inflammation and disease and to evaluate the efficiency of anti-inflammatory drugs in vivo.

STUDY OF THE EFFECT OF CRYOPRESERVED PLACENTA EXTRACT ON THE PROCESSES OF CYTOLYSIS AND LIPID PEROXIDATION IN CCL4-INDUCED LIVER DAMAGE

Abstract. The search for new strategies for the correction of exogenous toxic liver lesions is due to the steady increase in the incidence of hepatitis and cirrhosis among the working population, which is an important medical and social problem.
 The aim is to determine the effect of cryopreserved placenta extract (CEP) on the state of the liver of rats with tetrachloromethane (CCl4)-induced damage by indicators of lipid peroxidation (LP) and markers of cytolysis.
 Materials and methods. Experimental studies were conducted on 28 male rats. Acute CCl4-induced hepatitis was reproduced by a single injection of 50.0% CCl4 oil solution. KEP was administered 1 time per day for 5 days before the introduction of CCl4. The material for the study was whole blood and liver homogenates, in which the content of reactants with thiobarbituric acid (TBA-RP), catalase activity, superoxide dismutase (SOD) activity, alanine aminotransferase (AlAt) and aspartate aminotransferase (AsAt) activity, as well as γ-glutamyl activity were determined. γ-glutamyl transpeptidases (γ-GTP) and alkaline phosphatase (AP) according to standard methods.
 Results and discussion. The study showed that the content of TBA-RP in liver homogenates was lower (p<0.01) by 35.6% in rats that were prophylactically injected with CEP compared to rats with simulated CCl4-induced hepatitis without treatment (control group). An increase in the level of catalase (p=0.02) with the use of CEP was established by 33.8% and an increase in the activity of SOD (p<0.01) by 45.5% compared to the indicators of rats in the control group. It is also shown that the level of AlAt after administration of CEP decreased (p<0.001) by 56.0%, the level of AsAt decreased (p<0.001) by 48.6%, the level of γ-HTP decreased by 37.8% compared to the rats with untreated CCl4-induced hepatitis.
 Conclusions. Prophylactic five-day administration of CEP leads to the leveling of CCl4-induced LP activation and signs of cytolysis syndrome.

Regenerative effect tissue engineering scaffold based on fibrous permeable TiNi-based alloy

The aim of this study was to explore the in vivo applicability of intracorporeal constructions with hepatocytes cultured in the fiber TiNi-based scaffold (FTNS). We also sought to determine whether application of hepatocytes cultured in the FTNS was superior to simple hepatocyte injection in terms of restoration of liver functions and survival time in a CCl4-induced hepatitis rat model. In investigation 200 animals were assigned to explore the experimental groups as follows: Group I – control; Group II – animals with CCl4-induced hepatitis; Group III – animals with toxic hepatitis followed by implantation of cell-free FTNS (sham-surgery); Group IV – animals with toxic hepatitis followed by infusion of hepatocytes only (20 × 106 cells/ml); Group V – animals with toxic hepatitis followed by implantation of hepatocytes cultured in the FTNS. In terms of restoration of liver function, Group V showed a significant reduction of serum alanine aminotransferase level compared to Group II. Group IV also showed a significant decrease in ALT level at 15 days. However, the level of ALT increased at 30 days, and the level was similar with results of the Group II. This probably was caused by a short-term effect of hepatocyte injection only. The change patterns of serum protein, lactate, albumin, fibrinogen, and total bilirubin levels were similar to the results of ALT. The survival time of animals was significantly longer in Group V. These findings showed possible abilities of the FTNS as a scaffold to support the hepatocellular metabolism. Using 12 animals, image analysis of SEM data of in vivo hepatocyte evolution in the FTNS was carried out. In the SEM analysis, hepatocytes demonstrated good adhesion and proliferation in the pore space of the FTNS. Moreover, mature tissue comprising both colloidal and fibrous components filled the pore body by 95% in 28 days. Overall, this study sheds a light on how the implantable «auxiliary liver» by engrafting the cultured FTNS substitutes the missing hepatic function without the need to replace the whole liver.

Nitric oxide-mediated effects of L-ornithine-L-aspartate in acute toxic liver injury

This study was aimed to investigate nitric oxide-dependent mechanisms of L-ornithine-L-aspartate (LOLA) action in acute toxic liver injury in rats. Acute hepatitis was induced in Wistar rats using 50% oil solution of tetrachloromethane (CCl4) intragastrically (2 g/kg) twice in a 24 hour interval. Intraperitoneal treatment with LOLA (200 mg/kg) was started 6 hours after the second CCl4 administration and maintained for 3 consecutive days. L-Nω-Nitroarginine Methyl Ester (L-NAME) was used intraperitoneally (10 mg/kg). In CCl4-induced hepatitis, LOLA restores the structure of hepatocytes and prevents aminotransferases, alkaline phosphatase and gamma-glutamyl transferase elevation. It decreases total bilirubin concentration but does not affect increased cholesterol level. LOLA augments urea concentration, total protein level in blood and liver as well as serum and liver content of nitrite anions. LOLA enhances activity of catalase, glutathione S-transferase, manganese superoxide dismutase, increases reduced glutathione level and total antioxidant capacity and decreases thiobarbituric acid reactive substances level. The concomitant use of L-NAME inhibits the action of LOLA to enhance nitrite anions synthesis both in serum and liver, to delay the recovery of hepatocytes, to counteract LOLA effect against blood total protein reduction, to prevent the decline in aminotransferases, alkaline phosphatase,, gamma-glutamyl transferase and glutathione S-transferase activity and to reduce catalase activity and reduced glutathione level. Therefore, in CCl4-induced hepatitis, LOLA effectively prevents cytolysis and cholestasis, improves liver metabolism and protects against oxidative stress. Partially, these changes occur in nitric oxide-mediated mechanism since the use of L-NAME declines most of LOLA effects.

Acute toxicity, hepato-curative activity of extracts of a combination of plants on CCL4-induced hepatotoxicity in rats and antiradical activity

Traditional healers combine four medicinal plants (Cochlospermum tinctorium, Terminalia macroptera, Leptadenia hastata and Commiphora Africana to treat hepatitis in Burkina Faso. The aimed was to evaluate the hepato-curative activity of lyophilized aqueous decoction (LAD) and hydroethanolic macerate (LHM) of plant extracts on CCl4-induced hepatitis in rats. We assessed the acute toxicity and scavenging activity of the 2, 2-Diphenyl-1-picrylhydrazyl (DPPH). Hepato-curative activity study included nine groups with five rats each. We used rats as followed: group 1 as neutral controls, group 2 as negative controls, and the other groups were experimental groups. Rats in groups 2-9 received a single dose (1 mL/kg) of CCl4 in intraperitoneal injection to induce hepatitis. We fed orally the rats for seven consecutive days with sylimarin in group 3, LAD and LHM respectively in groups 4-6 and groups 7-9 by 400, 200 and 100 mg/kg/day. This study revealed LAD and LHM had a LD50> 2000 mg/kg and both showed radical-scavenging properties with IC50= 5.95 and 8.66 µg/mL respectively. All experimental rats regardless of the treatment group showed a significantly reduced plasma transaminases level as compared to negative controls. LAD and LHM at 400, 200 mg/kg significantly reduced alkaline phosphatase and gamma-glutamyl transferase. Histologically, treated rats showed normal to almost normal liver in a dose-dependent manner as compared to the controls. Conclusion: LAD and LHM decreased liver enzyme and allowed a dose-Dependent liver damage recovery after CCl4-induced hepatitis in rats.

Biostimulants improve the hepatoprotection of Ammi visnaga seed yield extract against carbon tetrachloride induced acute hepatitis in mice through modulation of MAPK

BackgroundThe liver is a vital organ that must be maintained healthy. Therefore, hepatoprotective agents deserve a big concern and exploring. ObjectivesWe aimed to enhance the yielded seed extract of Ammi visnaga by kinetin (KN) seed priming or asparagine (ASN) foliar spraying to explore the possibility of enhancing the hepatopritective effect against CCl4-induced acute hepatitis in mice through modulation of MAPK pathway. MethodsA. visnaga seeds were primed with 300 mg/L KN and other seeds were sprayed foliarly with 200 mg/L ASN. The phytochemical and GC/MS analyses were conducted. Ethanolic extract was used to explore its hepatoprotective potential in mice that were grouped to five groups; normal control, CCl4 control, CCl4 + untreated A. visnaga seeds, CCl4 + KN A. visnaga seeds, and CCl4 + ASN A. visnaga seeds. Acute hepatitis was induced by CCl4. Liver enzymes, oxidative stress biomarkers, inflammatory cytokines, and MAPK proteins were assessed. ResultsKN- and ASN- treated seeds increased number of inflorescences, total phenolic compounds, antioxidant capacity over untreated seeds, but ASN surpassed KN. ASN produced new compounds that were not detected in the untreated seeds. Therefore, ASN remarkably decreased hepatotoxicity markers, oxidative stress biomarkers, inflammatory cytokines, and phosphorylated MAPK proteins with restoring of normal hepatic architecture compared to the untreated and KN seeds. ConclusionsFoliar application of ASN was superior to KN priming in increasing seed yield, enhancing the metabolomics profile of the produced seed yield, and improving the corresponding pharmacological activity mainly through suppression of MAPK pathway.

HEPATOPROTECTIVE EFFECT OF CRYOPRESERVED PLACENTA EXTRACT ON THE TETRACHLOROMETHANE LIVER INJURY

Introduction. Toxic lesions of the hepatobiliary system are an important medical and social problem due to the steady increase in the frequency of chemical hepatosis, which develops as a result of accumulation of various xenobiotics in the body. Tetrachloromethane (CCl4) is one of the most studied hepatotoxins. Biochemical and pathomorphological changes in the liver lesions modeled by the administration of CCl4 are analogous to acute hepatitis of various etiologies in humans. As a potential hepatoprotective agent, cryopreserved placenta extract (CPE) attracted our attention. The objective is to characterize the effect of cryopreserved placenta extract on the functional state of the liver of rats with tetrachloromethane-induced damage. Materials and Methods. In vivo study was conducted on 28 male rats weighing 200–220 g. Acute CCl4-induced hepatitis was reproduced by injecting a 50.0% oil solution of CCl4 at a dose of 10 ml/kg body weight of the animal once, which caused acute fatty liver dystrophy. Animals were sacrificed 24 hours after the administration of CCl4. CPE was administered once per day for five days before CCl4 administration. The research material was blood and liver homogenates. The concentration of bilirubin in blood was determined by the spectrophotometrical method. The content of adenosine monophosphoric, adenosine diphosphoric, and adenosine triphosphoric acids was studied in the deproteinized homogenate by chromatographic method. Results and discussion. It was established that the development of CCl4-induced hepatitis is accompanied by the formation of a cholestatic syndrome and a disorder of energy metabolism in liver tissues. This was indicated by an increase (p < 0.001) in the level of total bilirubin by 4.7 times and a decrease (p < 0.001) in the energy charge by 42.6% compared to the indicators of intact rats. Prophylactic five-day administration of CPE led to a decrease (p < 0.001) in the level of total bilirubin by 32.1% and a decrease (p < 0.001) in the levels of direct and indirect bilirubin by 10.6% and 65.1%, respectively, compared to the indicators of animals with experimental toxic hepatitis. This indicated the weakening of the cholestatic syndrome and, indirectly, the weakening of inflammatory infiltration against the background of cytolytic processes in the liver. CPE had an energy-stabilizing effect on hepatocytes of rats with simulated tetrachloromethane liver injury. This was indicated by an increase (p = 0.02) of the energy charge by 18.2% compared to the indicators in the control group. Conclusions. The study showed anticholestatic and energy-stabilizing activity of the cryoextract of placenta used as a prophylactic measure in the model of tetrachloromethane-induced hepatitis.

Monooxygenase System and NO Metabolism in Liver Microsomes of Rats with Toxic Hepatitis and the Effect of Sesquiterpene Lactones.

We analyzed changes in activities of enzymes of phases I and II of xenobiotic biotransformation and parameters of NO metabolism in liver microsomes of rats with toxic CCl4-induced hepatitis after a 14-day course of sesquiterpene lactones from Artemisia leucodes (10 mg/kg). It was found that toxic hepatitis was associated with significant inhibition of NADPH-cytochrome c-reductase, benzo(a)pyrene hydroxylase, and NADPH-diaphorase, reduced cytochrome P-450 content, and enhanced induction of nitrate/nitrite reductase with accumulation of NO metabolites in the liver. Administration of sesquiterpene lactones stimulated activity of the studied components of the cytochrome P-450 system and promoted recovery of the NOergic system components; the effects were most pronounced in 7 and 14 days after treatment.

Novel near-infrared fluorescence probe with large Stokes shift for monitoring CCl4-induced toxic hepatitis

Toxic hepatitis which is induced by chemical substance is a serious threat to human health. More and more studies have shown that peroxynitrite (ONOO−) is related with the development of toxic hepatitis. So it is important to find a tool to study ONOO− change during the diagnosis and therapy of toxic hepatitis. Herein, a series of novel near-infrared (NIR) fluorescence dyes (DDM-R) with long emission wavelength (740–770 nm) and large Stokes shift (~200 nm) are developed. Among the dyes, DDM-OH with great spectral performance and facilely modified feature is used to construct probe DDM-ONOO-. The probe have the preference of high sensitivity and excellent selectivity for ONOO−. In addition, DDM-ONOO- was applied in detecting exogenous and endogenous ONOO− in cells and further used in detecting ONOO− of CCl4-induced toxic hepatitis in cells by fluorescence imaging, 3D quantification analysis, flow cytometry. More importantly, by visualizing ONOO−, the probe was used to monitor the diagnosis of CCl4-induced toxic hepatitis in mice and evaluate the therapeutic efficacy of hepatoprotective medicines (NAC, SM, DDB). The results show that the probe will provide a powerful tool for the diagnosis and treatment of toxic hepatitis.

Correction of Toxic Liver Damage with a Multicomponent Herbal Extract in an Animal Experiment

Background: Digestive diseases constitute a significant part in the overall structure of human diseases. Herbal cholagogues are indicated for a treatment of chronic liver, gall bladder, and bile ducts diseases. The aim of the work is to determine the choleretic effect of a new multicomponent plant extract. Materials and Methods: Multicomponent plant extract was obtained from the following types of plant materials: 300 g of immortelle flowers (Helichrysum arenarium L.), 100 g of tansy flowers (Tanacetum vulgare L.), 100 g of rose fruits (Rosa sp.), 100 g of leaves of common nettle (Urtica dioica L.), 50 g of mint leaves (Mentha piperita L.), 50 g of licorice roots (Glycyrrhiza glabra L.). The extract was standardized by the total flavonoid content. It was calculated and expressed in terms of luteolin and isosalipurposide standards (total flavonoids content: not less than 4% and 15% respectively). The animal experiments being done in 80 nonlinear male rats with initial body weight 180-200 g. In order to study a choleretic effect of multicomponent herbal extract, naive rats recieved the single experimental dose of 250 mg/kg. Pharmacotherapeutic activity was studied in white rats with CCl4-induced hepatitis. Results: Studies indicate a pronounced choleretic effect of the studied plant extract, that is comparable with the effect of “Allochol” in intact rats experiments. The course administration of a per os (peroral) multicomponent plant extract in a dose of 250 mg/kg to white non-linear rats with tetrachloromethane liver damage has a choleretic effect: it increases the rate of bile secretion, stimulates the synthesis and secretion of cholates with bile, and also the excretion of cholesterol and bilirubin. Conclusion: The obtained research results argue the feasibility of using a multicomponent plant extract containing biologically active substances of phenolic nature in the prevention and comprehensive treatment of liver diseases.

Repair of Erythrocyte Membranes by the Lipid Fraction from Brown Seaweed Sargassum Pallidum after Experimental Ccl4-Induced Toxic Hepatitis

The influence of the lipid fraction from the aqueous EtOH extract of brown seaweed Sargassum pallidum thallus and the commercial reference drug Essentiale® on the physiological characteristics of rat erythrocytes and the lipid composition of erythrocyte membranes after CCl4-induced toxic injury was studied. CCl4 intoxication (50% solution in olive oil, s.c., 2 mL/kg for 4 d) was associated with changes of all studied physiological and biochemical erythrocyte parameters that did not return to normal after withdrawing the toxicant. Intragastric administration to rats through a probe of the lipid fraction from S. pallidum extract at a dose of 80 mg of total lipids per kg of mass for 7 d after CCl4 intoxication gave membrane protection that manifested as a restoration of the characteristic erythrocyte dimensions (average volume and diameter), extension of the threshold to their hemolysis stability, and retention of the lipid constituent ratio in the membrane. The lipid fraction from brown seaweed S. pallidum was comparable to Essentiale® for restoring erythrocyte physiological characteristics and the lipid composition of their membranes after CCl4-induced damage. The pronounced protective effect of the S. pallidum lipid fraction for repairing erythrocyte membranes was evidently determined by the combined action of the marine lipids in it.

Inhibition of oxidative stress and NLRP3 inflammasome by Saikosaponin-d alleviates acute liver injury in carbon tetrachloride-induced hepatitis in mice.

NLRP3 inflammasome activation results in severe liver inflammation and injury. Saikosaponin-d (SSd) possesses anti-inflammatory and hepatoprotective effects. This study aimed to determine the protective effects of SSd on carbon tetrachloride (CCl4)-induced acute liver injury in mice, and whether oxidative stress and NLRP3 inflammasome activation participate in the process.The CCl4 mice model and controls were induced. The mice were treated with SSd at 1, 1.5, or 2.0 mg/kg in a total volume of 100 µl/25 g of body weight. Liver injury was assessed by histopathology. Oxidative stress was determined using mitochondrial superoxide production (MSP), malondialdehyde (MDA) content, and superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) activities. NLRP3, ASC, and Caspase 1 were determined by real-time PCR and western blot. IL-1β and IL-18 levels were determined by ELISA.Significantly elevated oxidative stress was induced in the liver by CCl4, as demonstrated by histopathology and increases of MDA and MSP levels and decreases of SOD, GPx, and CAT activities (all P < 0.01). SSd significantly decreased the MDA and MSP levels and increased the activities of SOD, GPx, and CAT (all P < 0.05). The mRNA expression of NLRP3, ASC, and Caspase 1, and the protein expression of Caspase 1-p10, NLRP3, ASC, IL-1β, and IL-18 were significantly increased after CCl4 induction (all P < 0.01). These changes were reversed by SSd (all P < 0.05).Suppression of the oxidative stress and NLRP3 inflammasome activation were involved in SSd-alleviated acute liver injury in CCl4-induced hepatitis.

Xymedone conjugate with para-aminobenzoic acid. Estimation of hepatoprotective properties

A salt-like conjugate of xymedone with para-aminobenzoic acid in a series of pyrimidine derivatives was synthesized, and its hepatoprotective properties were studied. The compound studied exhibits a cytoprotective effect at a concentration of 25 μmol L-1 enhancing the viability of normal human hepatocyte cells of the Chang Liver line by 2.1 times against the background of the impact of the d-galactosamine toxicant, which was shown by experiments in vitro. The cytotoxicity of the compound (IC50) is 20.7 mmol L-1. The data indicating the hepatoprotective effect most pronounced at the early stages of therapy were obtained in experiments in vivo performed according to the therapeutic scheme on the model of CCl4-induced toxic hepatitis. The ability of the conjugate to exert reparative and protective effects was found, since the surface area of destructive-degenerative and necrotic injuries revealed in hematoxylin- and eosin-stained sections on the third day of intraperitoneal administration of the studied compound in a dose of 0.7 mg kg-1 decreased by 1.5 times. The areas of detection of lipid inclusions in frozen sections stained with Sudan black decreased by four times on the third day at a dose of 1.7 mg kg-1, whereas the decrease was 3.2 times on the seventh day at a dose of 0.7 mg kg-1 compared to the control group of animals administrated with saline. According to the biochemical parameters, positive effects on the secretory and synthetic functions of the liver, bilirubin metabolism, and metabolism of iron and magnesium were observed during the treatment of animals with the conjugate.

Hepatoprotective Activity of Betulin and Dipterocarpol Derivatives

The regioselective synthesis of betulin 3,28-bis- and 28-monoacylates (nicotinate and isonicotinates) as well as of 3β-hemisuccinyl dipterocarpol was carried out using the dicyclohexylcarbodiimide method. The morphological and biochemical blood tests on a model of CCl4-induced rat hepatitis showed that the activities of the betulin bisacylates 3β,28-bis-О-nicotinate and 3β,28-bis-О-isonicotinate were higher than that of monoacylate 28-О-isonicotinate. 3β-Hydroxydipterocarpol was found to be the most active among triterpenoids of the dammarane type.

Hepatoprotective Effect of Melatonin in Toxic Liver Injury in Rats.

Background and objectives: toxic liver injury results in nitrooxidative stress. Melatonin is a potent free radical scavenger, an inducible nitric oxide synthase (iNOS) inhibitor and an activator of antioxidant enzymes. The aim of this study was to investigate the hepatoprotective effect of exogenous melatonin on animals with acute toxic hepatitis. Material and methods: 36 healthy Sprague-Dawley male rats were split into three equal groups and given carbon tetrachloride (CCl4), 2 g/kg (CCl4 group) or the same dose of CCl4 and melatonin, 10 mg/kg (CCl4/melatonin group) or saline (control group). The effect of melatonin on prooxidant and antioxidant system indexes, NO and NOS levels in serum and liver, data of mitochondrial chain functions and cytolysis in liver were evaluated in all three groups. Results: melatonin significantly decreased activities of AST, ALT, ceruloplasmine and thiobarbituric acid reactive substance (TBARS) in serum. Catalase activity was lowered in serum but not in the liver. Hepatic TBARS, lipid hydroperoxides and glutathione concentrations were decreased, while superoxide dismutase, mitochondrial cytochrome oxidase and succinate dehydrogenase activities increased. Melatonin inhibited synthesis of stable NO metabolites in serum: NO2-by 37.9%; NO3-by 29.2%. There was no significant difference in content NO2-in the liver, but concentration of NO3-increased by 32.6%. Melatonin significantly reduced iNOS concentrations both in serum (59.7%) and liver (57.8%) but did not affect endothelial isoform enzyme activities neither in serum, nor in liver. The histopathological liver lesions observed in the CCl4/melatonin group were less severe than those seen in the CCl4 group. Conclusions: we demonstrated an ameliorating effect of melatonin on prooxidants and antioxidants, NO-NOS systems balance, mitochondrial function and histopathological lesions in the liver in rats with CCl4-induced hepatitis.

TO THE MECHANISM OF SEASONAL VARIATIONS IN CARBON TETRACHLORIDE TOXICITY

Introduction. Seasonal biorhythms can modify the toxic effect of xenobiotics. Natural fluctuations in hormone levels in the blood which can modify the metabolic processes in the liver can be the base of one of the mechanisms in this phenomenon. The study of the effector role of testosterone in seasonal chrono-reactivity can reveal one new link in the pathogenesis and will improve health risk assessment, especially in patients taking hormone therapy. The aim of the study was to reveal the role of testosterone in the mechanisms of chrono-reactivity the toxic effect for carbon tetrachloride in warm-blooded animals. Material and Methods. Studies were conducted on adult male Wistar rats, divided into negative control groups, a model with CCl4-induced toxic hepatitis, and experimental in which CCl4 exposure was carried out following a preliminary 7-day course of testosterone administration. Direct and total bilirubin, cholesterol, alkaline phosphatase, AlAT and AsAT, hydroperoxide and malondialdehyde were determined by standard methods. The experiment was repeated twice: both in winter and spring. Results. The amplitude of the damage to the structures and functions of hepatocytes from a single injection of the same poison dosage in winter and spring manifested in varying degrees, revealing the modulatory role of biological rhythms. Under premedication with testosterone, a similar tendency was observed in both series of the experiment (winter, spring): the toxic effect of CCl4 increased. This was recorded by the following biochemical changes: bilirubin increased by 55%, cholesterol - 19%, alkaline phosphatase - 12%, AlAT - 17%, AsAT - 35%, hydroperoxide - 29%, and malonic dialdehyde slightly (relative to groups with model toxic hepatitis). Conclusion. The male sex hormone regulating the metabolic activation of cytochromes in hepatocytes performing biotransformation is one of the effector links in the seasonal chrono-reactivity phenomena. The discovery of the whole mechanism will allow developing an effective system of chronoprophylaxis and reducing the risks of toxic poisoning in individuals during periods of maximum chrono-reactivity.

THE SEASONAL VARIATIONS OF CARBON TETRACHLORIDE TOXICITY

Biological rhythms are genetically fixed as a universal form of adaptation to cyclically changing environmental factors. Many organs including the liver and body systems function in an oscillatory mode. Its central role in the process of detoxification may determine the variability of toxic effects in the metabolism of xenobiotics. The including of chronobiological patterns in the procedures of toxicological and hygienic studies will allow more accurate assessing of the properties of tested substances. The aim of the study was to identify variations in the seasonal chrono-reactivity of warm-blooded animals to the hepatotoxic effects of carbon tetrachloride. In studies there were used Wistar rats, divided into experimental (CCl4-induced toxic hepatitis) and control groups in the winter, spring, summer, and autumn. Standard laboratory diagnostic methods studied key indices characterizing the functional activity of the liver and the antioxidant status of the organism (direct and total bilirubin, cholesterol, alkaline phosphatase, ALAT, ASAT, hydroperoxide, malonic dialdehyde, catalase in the liver tissues). According to the results of the conducted research, seasonal biorhythms have a modulating effect on the organism of warm-blooded animals, and the amplitude of the detected fluctuations was observed to be wider in terms of modeled pathology. The calculation of the variation values of numerical indices shows fluctuations to take place in the range of 10-60% in control group, and within the range of 60-300% in the pathology. The most labile index was catalase. Since the mechanism of carbon tetrachloride toxicity is mediated through the initiation of lipid peroxidation, this may explain the observed phenomena of seasonal chronoreactivity. Thus, taking into account the modulating role of biorhythms in protocols for toxicity testing can optimize the currently used procedures of human health risk assessment.

Protective Effect of Antrodia cinnamomea Extract against Irradiation-Induced Acute Hepatitis

Radiotherapy for treatment of hepatocellular carcinoma causes severe side effects, including acute hepatitis and chronic fibrosis. Complementary and alternative medicine (CAM) has emerged as an important part of integrative medicine in the management of diseases. Antrodia cinnamomea (AC), a valuable medicinal fungus originally found only in Taiwan, has been shown to possess anti-oxidation, vaso-relaxtation, anti-inflammation, anti-hepatitis, and anti-cancer effects. In this paper we evaluate the protective effects of ethanol extract of Antrodia cinnamomea (ACE) against radiotoxicity both in normal liver cell line CL48 and in tumor-bearing mice. In CL48, ACE protects cells by eliminating irradiation-induced reactive oxygen species (ROS) through the induction of Nrf2 and the downstream redox system enzymes. The protective effect of ACE was also demonstrated in tumor-bearing mice by alleviating irradiation-induced acute hepatitis. ACE could also protect mice from CCl4-induced hepatitis. Since both radiation and CCl4 cause free radicals, these results indicate that ACE likely contains active components that protect normal liver cells from free radical attack and can potentially benefit hepatocellular carcinoma (HCC) patients during radiotherapy.