Content of Mineral Elements in Rat Liver Tissue with Acquired Insulin Resistance and Obesity in Combination with Iodine

The purpose of the study was to determine the content of mineral elements in the hepatocytes of rats with insulin resistance, obesity, iodine deficiency, insulin resistance in combination with iodine deficiency and obesity in combination with iodine deficiency. Materials and methods. The study was performed on 90 white nonlinear rats weighing 120-180 g, which were divided into five experimental groups: rats with insulin resistance (1st experimental group, n = 15), animals with iodine deficiency (2nd experimental group), animals with insulin resistance in combination with iodine deficiency (3rd experimental group, n = 15), obese animals (4th experimental group, n = 15), obese animals in combined with iodine deficiency (5th experimental group, n = 15). The control group consisted of 15 intact rats. The content of Fe, Ca, Cu, Mg, Zn, Mn, Cr in the liver homogenate was determined by atomic absorption spectrometry on a SPECORD M 40 spectrophotometer (Germany). Results and discussion. In animals with insulin resistance, there was a decrease in the content of copper in the liver by 26.9%, in animals with iodine deficiency the content of this trace element increased by 20.5%, and in the group of animals with insulin resistance in combination with iodine deficiency it increased by 10.1%. The iron content in the group of animals with insulin resistance is higher in relation to the control by 33.7%, in the group of animals with iodine deficiency – by 38.5%, in animals with insulin resistance in combination with iodine deficiency – by 40.8%. Regarding the content of Calcium, in the liver homogenate of animals with insulin resistance it is higher compared to the control by 24%, in animals of the second experimental group –by 26.4%, in animals of the 3rd experimental group – by 22%. The Magnesium content in animals with insulin resitance is lower compared to the control by 19%, in animals with iodine deficiency – by 25.5%, in animals with insulin resistance in combination with iodine deficiency – by 29%. As for Zinc, no significant fluctuations in the content of this trace element were detected. In animals of the 1st, 2nd and 3rd experimental groups it is lower compared to the control by 10%, 15% and 11.1%, respectively. The Manganese content in animals with insulin resistance is lower compared to the control by 13.6%, in animals with iodine deficiency – by 18.2%, in animals with insulin resistance in combination with iodine deficiency – by 14.8%. With regard to chromium, we found a probable decrease in the concentration of this trace element in animals of the group with insulin resistance by 48%, in animals of the 2nd experimental group – by 57.5%, in animals of the 3rd experimental group – by 58%. In the group of animals with obesity and obesity in combined with iodine deficiency we found an increase in copper content compared to the control by 27.4% and 36.2%, respectively. The iron content in animals with obesity exceeds the control by 34.8%, in the group of animals with obesity in combined with iodine deficiency – by 38.4%. Regarding the content of Calcium, in animals with obesity it is higher by 25.7%, and in animals with obesity in combined with iodine deficiency – by 28.4% compared to the control. Magnesium content in animals with obesity is lower by 27.3% compared to the control group, and in animals with obesity in combined with iodine deficiency – by 28.4%. Regarding Zinc, no significant fluctuations in the content of this trace element were detected. In animals of the 4th and 5th experimental groups it is lower compared to the control by 18.4% and 23.5%, respectively. The content of Manganese in the group of animals with obesity decreased by 14.8%, and in the group with obesity in combined with iodine deficiency the content of this trace element decreased by 16.2% compared to the control. With regard to chromium, we found a probable decrease in the concentration of this trace element in animals of the group with obesity by 58.1% and in the group of animals with obesity in combined with iodine deficiency by 56.2%. Conclusion. The obtained results demonstrate changes in the content of mineral elements in groups of animals with insulin resistance, iodine deficiency, insulin resistance in combination with iodine deficiency, obesity and obesity in combination with iodine deficiency

The state of pro- and antioxidant systems in rats with DMH-induced colon carcinogenesis on the background of extracorporeal detoxification

Aim: Cancer is one of the leading causes of death in the world. The aim of this research was to study the indices of pro- and antioxidant systems in rats with dimethylhydrazine (DMH)-induced colon carcinogenesis on the background of the enterosorbent AUT-M use. Materials and methods: The study was performed on 70 white male rats weighing 200–250 g. Adenocarcinoma of the colon was simulated by subcutaneous injection of the DMH (Sigma-Aldrich Chemie, Japan) at a dose of 7.2 mg/kg once a week during 7 months. Enterosorbent AUT-M was administered intragastrically daily for 21 days after simulation of carcinogenesis at a dose of 1 ml of suspension per 100 g of animal body weight. The state of the pro- and antioxidant systems was studied by the content of oxidative modification of proteins products (OMP), the activity of superoxide dismutase (SOD), catalase (CAT), contents of ceruloplasmin (CP) and reduced glutathione (GSH). Results: It was found that DMH-induced colon carcinogenesis in rats is accompanied by disorders in the antioxidant defense system and activation of free radical oxidation processes. Enterosorbent AUT-M provides a significant reduction in the content of OMP370 and OMP430 in both blood serum and liver homogenate of rats. Moreover, the use of enterosorbent AUT-M demonstrated a significant increase in the activity of SOD, CAT, content of GSH and a decrease in CP content in investigated tissues. Conclusion: The use of enterosorbent AUT-М demonstrated prominent potential suppression for oxidative stress and positive effect on antioxidant defense system in rats with DMH-induced colon carcinogenesis.

Study of hepatoprotective effect of bearberry leaves extract under insulin resistance in rats

The aim of our study was to evaluate the antidiabetic and hepatoprotective efficacy of dry extract from bearberry leaves enriched with arginine in dexamethasone induced IR. Materials and methods. IR was induced in rats by low dose intraperitoneally injections of dexamethasone. Dexamethasone-induced IR in rats was treated by bearberry leaves extract enriched with arginine. Thus, animals were randomized into several groups including intact animals and animals, which administered reference compounds and medications. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamine transferase (GGT) were determined in blood serum and liver homogenate, in addition, in blood serum we measured lactate dehydrogenase (LDH) activity and lactate level and glycogen content liver tissue. Also, for the purpose of our experiment, in liver tissue were determined: thiobarbituric acid reactive substances (TBARS), diene conjugates (DC), and reduced glutathione (GSH) content; and superoxide dismutase (SOD), glutathione peroxidase (Gpx), and catalase (CAT) activities. All indices were determined using generally accepted unified methods or commercially available kits. Results. Long-term dexamethasone administration led to an increase in AST, ALT and GGT overall activity in the liver homogenate and serum; this could be the result of increased permeability of hepatocyte plasma membranes, as well as their enhanced synthesis in the liver. Studied extract ameliorate these indices of liver injury. Evaluation of indices that reflected oxidative stress and the antioxidant system status in liver confirmed oxidative stress development in IR rats` liver. Administration of arginine enriched bearberry leaves extract decrease TBARS and DC content in liver tissue, at the same time, improve SOD, Gpx, and CAT activities and increase GSH content. Conclusions. Bearberry leaves dry extract enriched with arginine inhibit oxidative stress development, improve membrane integrity, and normalize some indices of carbohydrate metabolism, particularly glycogen content in liver and lactate level in blood.

Amelioration of Radiation-Induced DNA Damage and Immunosuppression by Tanacetum parthenium leaf extract and synthetic parthenolide in Swiss Albino Mice

Radiotherapy potentially offers protection from recurrence of tumor that also causes normal tissue damage and creates major concern. Another important factor is long-term immune suppression in patients treated with radiotherapy. Therefore, crucial need for the survival of surrounding normal cells of tumor by radiation‑protecting agents is the prime focus of this study. Aqueous extract (AE) and ethanolic extract (EE), Tanacetum parthenium extracts100 mg/kg each and parthenolide (PAR) 4mg/kg body weight were orally administered prior to sub-lethal radiation dose exposure. Mice were used for the evaluation of radiation-mediated chromosomal aberrations in bone marrow cells and DNA break by comet assay in the blood lymphocytes of mice. The pro-inflammatory levels were determined by cytokine estimations namely interleukin‑2, interferongamma and tumor necrosis factor-alpha performed in the liver homogenate using ELISA kits. Thus the results demonstrated ameliorating, radio-mitigating and immune-stimulatory efficacy of AE, EE and PAR against radiation ‑induced DNA damage and immunosuppression by regulating cytokine.

INFLUENCE OF THICK EXTRACT FROM MAITAKE MUSHROOMS ON SIGNS OF INFLAMMATORY PROCESS IN EXPERIMENTAL TOXIC HEPATITIS

Background. The priority of the contemporary pharmaceutical industry is to create effective, safe and inexpensive drugs to ensure the highest quality of care and optimal use of available raw materials. Objective. The aim of our study was to investigate anti-inflammatory properties of the Maitake mushrooms thick extract in the experiment on rats with paracetamol(acetaminophen)-induced hepatitis. Methods. 60 white male rats, weighing 180-210 g, randomized into 10 groups of 6 animals in each, were used for the experiment. Paracetamol hepatitis was simulated by acetaminophen intragastric administering in a dose of 1250 mg/kg 1 time per day (for 2 days) as a suspension in 2% starch gel solution. Maitake mushrooms thick extract, which was administered intragastrically 2 hours before the administration of acetaminophen and daily after the lesion in a dose of 150 mg/kg of the animal’s body weight, was used for the toxic lesion correction. “Silibor” was selected as the comparison drug, which was administered according to the same scheme as the investigated extract in a dose of 20 mg/kg of the animal’s body weight. Euthanasia was conducted on the 3rd, 7th and 10th day of the experiment with sodium barbamyl. Liver homogenate and animal serum were used for the studies. The development of inflammatory processes was studied by the content of pro-inflammatory and anti-inflammatory cytokines, as well as C-reactive protein in the serum of rats with toxic hepatitis and after the application of Maitake mushroom extract and the comparison drug. Results. It was found that the introduction of acetaminophen to animals for the acute hepatitis simulation is accompanied by changes in the cytokine profile, i.e. an increase in the level of IL-6 and a decrease in the level of IL-4 in the serum of rats. Inflammatory development is evidenced by the content of C-reactive protein increase in the blood of the affected animals. The application of Maitake mushroom extract facilitated bringing the studied indicators almost to the level of intact control. Conclusions. Reduction of inflammation signs in rats with the simulated paracetamol hepatitis under the influence of Maitake mushrooms thick extract confirms its anti-inflammatory properties. Objective. The aim of our study was to investigate anti-inflammatory properties of the Maitake mushrooms thick extract in the experiment on rats with paracetamol(acetaminophen)-induced hepatitis. Methods. 60 white male rats, weighing 180-210 g, randomized into 10 groups of 6 animals each, were used for the experiment. Paracetamol hepatitis was simulated by acetaminophen intragastric administering in a dose of 1250 mg/kg 1 time per day (for 2 days) as a suspension in 2% starch gel solution. Maitake mushrooms thick extract, which was administered intragastrically 2 hours before the administration of acetaminophen and daily after the lesion in a dose of 150 mg/kg of the animal’s body weight, was used for the toxic lesion correction. "Silibor" was selected as the comparison drug, which was administered according to the same scheme like the investigated extract in a dose of 20 mg/kg of the animal’s body weight. Euthanasia was conducted on the 3rd, 7th and 10th day of the experiment with sodium barbamyl using. Liver homogenate and animal serum were used for the studies. The development of inflammatory processes was studied by the content of pro- and anti-inflammatory cytokines, as well as C-reactive protein in the serum of rats with toxic hepatitis and after the application of Maitake mushroom extract and the comparison drug. Results. It was found that the introduction of acetaminophen to animals for the acute hepatitis simulation is accompanied by changes in the cytokine profile, namely, an increase in the level of IL-6 and a decrease in the level of IL-4 in the serum of rats. The inflammatory process development is evidenced by the content of C-reactive protein increasing in the blood of affected animals. The application of Maitake mushroom extract helped to bring the studied indicators closer to the level of intact control. Conclusions. The application of the Maitake mushrooms thick extract as a corrective factor at the simulated paracetamol hepatitis confirms its anti-inflammatory properties. KEYWORDS: Maitake mushrooms, paracetamol, hepatitis, inflammatory processes, thick extract, anti-inflammatory properties.

Hepatoprotective Effects of Calotropis procera (Ait.) R. Br Root Bark Extracts against Diethylnitrosamine Induced Liver Injury in Rats

Background: Diethylnitrosamine (DEN) is a hepatotoxin whose metabolic activation by liver cytochromes P450 is responsible for the necrosis, mutagenicity and carcinogenicity of liver cells. The purpose of this study was to evaluate the protective effects of Calotropis procera roots bark against DEN induced hepatocellular damage in rats. Material and Methods: Hepatoprotective activity of the ethanolic extract of Calotropis procera root bark were evaluated by induction of liver injury with DEN in Wistar male rats distributed in six groups of six. Serum hepatic markers, alanine amino transferase (ALAT), aspartate amino transferase (ASAT), alkaline phosphatase (ALP), total protein and albumin were evaluated and the enzymes antioxidant activities, superoxide dismutase (SOD) and catalase, as well as the level of malonedialdehyde (MDA) were determined in the liver homogenate. Histological analysis was carried out on sections of rat livers. Phytoconstituents have also been studied. Results: Pretreatment of rats with the extract showed a significant decrease in ALAT, ASAT and ALP while there was an increase in total protein and albumin compared to rats treated only with DEN. It also showed a significant increase in SOD and catalase and a decrease in MDA levels suggesting the hepatoprotective effect of the extract. Observation of liver sections confirmed the results of the biochemical parameters which would attest that the extract is hepatoprotective. Phytoconstituents such as sterols, triterpenes and phenolic compounds have been demonstrated. Conclusion: Ethanolic extract of Calotropis procera roots bark has shown hepatoprotective effects that could be due to its content in sterols and triterpenic and phenolic compounds.

ANALYSIS OF OXIDATIVE STRESS MARKERS IN CHRONIC CONSUMPTION OF MONOSODIUM GLUTAMATE ON LIVER OF WISTAR ALBINO RATS

Objective: The study was intended to explore whether Monosodium glutamate (MSG) induces oxidative stress on the liver of Wistar albino rats when fed chronically at three different doses, namely, low, mid, and high doses identical to human consumption doses in growing countries. Methods: The acclimatized Wistar albino rats (n=24) were randomly selected and grouped into four groups, namely Control, Low dose MSG (180 mg kg), Mid dose MSG (360 mg/kg), and High dose MSG (720 mg/kg). The animals were orally administered MSG for 120 days. After completion of the experimental period (120 days), euthanized animal liver was homogenized to investigate the oxidative stress marker enzymes such as Superoxide Dismutase (SOD), Glutathione Peroxidase (GPx), Catalase (CAT), and Myeloperoxidase (MPO). Results: The MPO showed a significant increase (p<0.05) in liver homogenate of all MSG induced groups when compared to control group. The SOD, CAT, and GPx activity deteriorated (p<0.05) in monosodium induced groups contrasting to the control group. Conclusion: The effects of MSG on oxidative stress markers on liver homogenate in the current study exhibited erratic abnormal changes in oxidative stress markers of monosodium induced groups which contemplate the harmful effects of MSG consumed chronically. The further studies should confirm the genetic basis of oxidative stress damage and transform the safety regulations of MSG consumption throughout the world.

Hepatic Ischemia-Reperfusion Impairs Blood-Brain Barrier Partly Due to Release of Arginase From Injured Liver

Aim: Hepatic ischemia-reperfusion (HIR) induces remote organs injury, including the brain. The homeostasis of the brain is maintained by the blood-brain barrier (BBB); thus, we aimed to investigate whether HIR impaired BBB and attempted to elucidate its underlying mechanism.Methods: Cell viability of human cerebral microvascular endothelial cells (hCMEC/D3) was measured following 24 h incubation with a serum of HIR rat undergoing 1 h ischemia and 4 h reperfusion, liver homogenate, or lysate of primary hepatocytes of the rat. The liver homogenate was precipitated using (NH4)2SO4 followed by separation on three columns and electrophoresis to identify the toxic molecule. Cell activity, apoptosis, proliferation, cell cycle, and expressions of proteins related to cell cycle were measured in hCMEC/D3 cells incubated with identified toxic molecules. HIR rats undergoing 1 h ischemia and 24 h reperfusion were developed to determine the release of an identified toxic molecule. BBB function was indexed as permeability to fluorescein and brain water. Endothelial cell proliferation and expressions of proteins related to the cell cycle in cerebral microvessels were measured by immunofluorescence and western blot.Results: Toxic molecule to BBB in the liver was identified to be arginase. Arginase inhibitor nor-NOHA efficiently attenuated hCMEC/D3 damage caused by liver homogenate and serum of HIR rats. Both arginase and serum of HIR rats significantly lowered arginine (Arg) in the culture medium. Arg addition efficiently attenuated the impairment of hCMEC/D3 caused by arginase or Arg deficiency, demonstrating that arginase impaired hCMEC/D3 via depriving Arg. Both arginase and Arg deficiency damaged hCMEC/D3 cells by inhibiting cell proliferation, retarding the cell cycle to G1 phase, and downregulating expressions of cyclin A, cyclin D, CDK2, and CDK4. HIR notably increased plasma arginase activity and lowered Arg level, increased the BBB permeability accompanied with enhanced brain water, and decreased the proliferative cells (marked by Ki67) in cerebral microvessels (marked by CD31) and protein expressions of cyclin A, cyclin D, CDK2 and CDK4 in isolated brain microvessels. Oral supplement of Arg remarkably attenuated these HIR-induced alterations.Conclusion: HIR leads to substantial release of arginase from the injured liver and then deprives systemic Arg. The Arg deficiency further impairs BBB via inhibiting the proliferation of brain microvascular endothelial cells by cell cycle arrest.

Effects of high-fat diets on inflammation and antioxidant status in rats : comparison between palm olein and olive oil

Palm olein (PO) and olive oil (OO) are widely consumed in the world. PO is considered harmful to health, whereas OO is considered healthy. The aim of the study was to compare the effects of consumption of these oils on antioxidant status and inflammation in rats. This was an experimental study in male wistar rats fed a diet containing 30% of each oil. Rats had free access to food and water. After being fed for 12 weeks, animals were sacrificed and liver and aortic blood were collected. Plasma was used for the determination of interleukin-6 (IL-6) and oxidative stress parameters (Superoxide dismutase -SOD; Gluthation peroxidase - GPx; Thiobarbituric acid reactive substances - TBARS; Thiol groups and isoprostane). The inflammation and oxidative stress status as well as the expression of several genes/proteins were also analyzed in liver homogenate. No significant differences were observed between PO and OO in plasma and liver levels of the studied inflammation and oxidative stress parameters. This study showed that the consumption of PO induces an antioxidant status superimposable to that of OO. Key words : Palm olein - Olive oil - Oxidative stress - Inflammation - High fat diet

Hepatotoxicity induced by intravenous administration of PEGylated nano-graphene oxide in albino mice

Graphene oxide (GO) has been intensely investigated in recent years due to its biocompatibility and its role in drug delivery. Its conjugation with polyethylene glycol (PEG) further improves its solubility in physiological solutions, which is important for enhancing efficacy of drug delivery. The present study aimed to assess the hepatotoxicity of PEG-nGO in mature mice. Liver function tests such as Alanine transferase (ALT), Alkaline phosphatase (ALP) were performed in the liver homogenate of the control and treated groups after intravenous administration of a single dose (5 mg/kg) of PEG-nGO through the tail vein. Total Glycogen content and lactate dehydrogenase (LDH) activity was measured. For histology studies, liver slices were fixed in 10% formalin and stained with H&E and photographed. The liver function test indicated a significant increase in ALT and ALP activity following 1 to 2 h of treatment with PEG-nGO, which recovers to normal levels at 4 h. Total glycogen contents were mobilized from the liver in the first hour in response to stress, which again regain normality after 4 h. The LDH assay showed maximum necrosis and apoptosis of hepatocytes at 1 h. Histology studies further indicated that infiltration of inflammatory cells and vacuolization of cytoplasm occurred mostly at 1 h. PEG-nGO treatments caused maximum damage and toxicity to the liver during the first 2 h. Following this, the liver tissues recover substantially which indicated that the low dose toxicity of PEG-nGO to the liver was transient and reversible.