Liver Of Wistar Rats Research Articles

Protective potential of Momordica. charantia against CCl4 induced change in liver and kidney serum biomarkers in Wistar rats

Protective potential of Momordica. charantia against CCl4 induced change in liver and kidney serum biomarkers in Wistar rats

The effect of aqueous extract of Lonchocarpus cyanencens leaf on the histology of the liver of Wistar rats

The effect of aqueous extract of Lonchocarpus cyanencens leaf on the histology of the liver of Wistar rats

The biochemical effects of Kola Nitida on the Liver of Lipid-Induced Wistar Rats

This study investigates the biochemical effects of Kola Nitida on the liver of Wistar rats subjected to a high lipid diet. A High lipid diet induces oxidative stress, inflammation, and pancreatic dysfunction. This study evaluated whether Kola Nitida, known for its bioactive compounds such as flavonoids, caffeine, and theobromine, could protect the liver from lipid-induced damage. The study was conducted within 6 months (April – September 2024). The effect of bioactive compounds from kola nut extracts on liver functions of Wistar rats was investigated using standard methods. Twenty Wistar rats were used for the study. They were randomly divided into five groups 1, 2, 3, control, and toxicity made up of 4 rats each. The control group received water food, Group 1 received kola nitida extract 100mg/kg body weight water food, group (B) received kola nitida extract 200mg/kg body weight water food, and Group (C) received kola nitida extract 300mg/kg body weight water food by oral administration for 14 days. The results showed that caffeine from kola nuts had effects on the liver functions of the Wistar rats. Administration of kola nitida (vera) significantly decreased aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), GGT, Total bilirubin (TB), Total protein (TP), Albumin (ALB), activities, consumption of HFD increased total bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP)activity. Body weight, organ weight, and histopathological study were significantly reduced. This study seems to suggest that consumption of HFD and reversal with kolanut (kola nitida) will significantly cause positive effects on the biological and histological liver functions of humans.

Phytochemical analysis, antioxidant activity and modulatory effect of aqueous leaf extract of Pistacia lentiscus L on oxaliplatin-induced oxidative stress in isolated rat liver mitochondria

This study evaluates the antioxidant effects of aqueous leaf extract of Pistacia lentiscus (ALEPL) and its potential to counteract oxaliplatin (OXA)-induced mitochondrial oxidative stress in rat livers, a common side effect of chemotherapy in cancer treatment. Bioactive compounds were identified using High-Performance Liquid Chromatography coupled with Tandem Mass Spectrometry (HPLC-MS and MS), with Fourier-Transform Infrared Spectroscopy (FTIR) and Atomic Absorption Spectrophotometry (AAS) for chemical and mineral analysis. ALEPL showed notable antioxidant activity, with IC50 values of 4.30 ± 0.27 μg/mL for DPPH (2,2-Diphenyl-1-picrylhydrazyl) scavenging, 13.64 ± 0.51 μg/mL for reducing power, 32.62 ± 5.32 μg/mL for hydroxyl radical scavenging, and 205.08 ± 25.77 μg/mL for superoxide anion radical scavenging. In ex vivo experiments, mitochondria isolated from Wistar rat livers were treated with OXA and ALEPL in a dose-dependent manner. ALEPL pretreatment effectively restored mitochondrial antioxidant enzyme activities, increased glutathione (GSH) levels, and reduced lipid peroxidation (MDA) caused by OXA. These findings suggest that ALEPL has the potential to act as a natural antioxidant to support cancer treatment by mitigating chemotherapy-induced oxidative stress. Future studies could explore its application as an adjuvant in clinical settings to enhance the efficacy of chemotherapy while reducing its side effects.

Biochemical Analysis of Persea americana Extract for Mitigating Cadmium-Induced Liver Damage in Rats

This research investigated the effects of methanol seed extract of Persea americana Mill on the liver of Wistar rats exposed to cadmium toxicity. The liver, known for its critical role in metabolism and interaction with the gastrointestinal tract, is predisposed to damage caused by drugs and other harmful chemicals. According to research, P. americana is rich in secondary metabolites with hepatoprotective properties. The seeds of P. americana were obtained from its fruits, procured from a local seller. The seeds were cut, air-dried under shade until crisp, pounded into powder, and extracted in methanol for 72 hours. Exactly 20 rats were subdivided into four groups, each containing five rats. The normal control group was given normal saline. The second group received 20 mg/kg bw CdSO4, the third group received 20 mg/kg bw CdSO4 with 400 mg/kg bw extract, and the fourth group received 20 mg/kg bw CdSO4 with 100 mg/kg bw silymarin. The results showed a significant difference in AST, ALT, ALP, and GGT levels in group 1 compared to the second group, which exhibited an increase. The treated groups demonstrated a considerable decrease in these marker enzymes relative to the induced group. There was no statistically significant difference in the protein levels among the groups. However, albumin levels decreased in group 2 compared to the other groups. MDA levels increased significantly in group 2, while SOD and CAT levels showed a significant decrease in group 2. The plant extract exhibited a promising hepatoprotective effect on the liver of animals induced with cadmium (CdSO4).

Green synthesis of silver nanoparticles containing Cichorium intybus to treat the sepsis-induced DNA damage in the liver of Wistar albino rats

Abstract Sepsis is a severe reaction of the body to an infection, presenting a critical medical crisis. It represents an imbalance between the body’s anti- and pro-inflammatory reactions. The occurrence of sepsis, which leads to multiple-organ failure and increased mortality, is marked by dysfunction in the lungs, heart, kidneys, and liver. The involvement of reactive oxygen species is believed to contribute to the progression of sepsis. Data suggest potential advantages of phenolic compounds derived from plants in combating sepsis. Plant polyphenols can be antioxidants by scavenging free radicals, chelating metals, and binding to proteins. In this research, silver nanoparticles (AgNPs) were produced by the aqueous extract of Cichorium intybus leaf for the purpose of treating sepsis-induced DNA harm. The recent study focused on the biological aspect including the cytotoxicity properties on normal (HUVEC) cell line. The AgNPs were analyzed by transmission electron microscopy (TEM), scanning electron microscopy (SEM), fourier-transform infrared spectroscopy, X-ray powder diffraction, and UV-Vis. The TEM and SEM images of AgNPs exhibited the average size of 35.29 nm with spherical morphology. In the in vivo study, the animals were categorized into four groups: sepsis-induced, sham, AgNPs-20, and AgNPs-100. AgNPs treatment resulted in a significant decrease in tissues damage (p < 0.01). The sepsis-induced group showed a significantly elevated malondialdehyde (MDA) level in comparison to the sham group (p < 0.01). Nevertheless, the groups that received AgNPs experienced a decrease in MDA levels and an increase in glutathione and superoxide dismutases levels (p < 0.01). Additionally, the rats treated with AgNPs exhibited a reduction in the IL-1β mRNA expression levels (p < 0.01).

The Effect of Copper Nanoparticles on Liver Metabolism Depends on the Type of Dietary Fiber.

A diet enriched with copper nanoparticles (CuNPs) exhibits a wide range of effects on liver metabolism, both positive and negative. Dietary fibers are the key components that may affect the absorption of minerals, including copper, and change their impact on organisms. Therefore, this study investigated whether and how supplementation with different sources of dietary fiber (cellulose, pectin, inulin, and psyllium) affects the function of CuNPs in the liver of male Wistar rats. The results showed that CuNPs at different doses had varying effects on lipid metabolism and inflammation in the liver. Specifically, higher doses of CuNPs were associated with increased lipid accumulation and the activation of pro-inflammatory mechanisms. However, combining CuNPs with dietary fibers, such as psyllium and inulin, was beneficial in mitigating the effects of the examined nanoparticles, leading to reduced fat, cholesterol, and triglycerides in the liver. Combining psyllium with CuNPs showed the most substantial effect on liver metabolism and inflammation parameters. Furthermore, hepatic histology analyses showed that adding psyllium to the diet with CuNPs reduces changes associated with fat accumulation and mononuclear cell infiltration. The observed beneficial changes in the liver may have been related to a reduction in the gene expression level of sterol regulatory element-binding protein 1 and peroxisome proliferator-activated receptor gamma and cyclooxygenase-2. In conclusion, enriching the diet with dietary fibers such as psyllium can regulate the action of CuNPs, thereby improving lipid metabolism and reducing inflammation in the liver.

Role of Callistemon citrinus Leaf Phytosomes Against Oxidative Stress and Inflammation in Rats Fed with a High-Fat-Fructose Diet.

Phytosomes are used as vehicles that carry plant extracts. They exhibit biological activities and possess better bioavailability, bioabsorption, and lower toxicity than drugs. Obesity is an inflammatory state in which oxidative stress is present, which triggers severe effects on the body's organs. This study aimed to evaluate the impact of the extract and phytosomes of Callistemon citrinus on oxidative stress and inflammation in the liver and heart of Wistar rats fed with a high-fat-fructose diet. Phytosomes containing the extract of leaves of C. citrinus were prepared. The antioxidant, pro-inflammatory enzymes, and biomarkers of oxidative stress were evaluated. Among the groups, only the high-fat-fructose group presented an increase in the COX-2, 5-LOX, and MPO inflammatory enzymes, while the XO enzyme exhibited decreased activity. The groups were fed a hypercaloric diet for 15 weeks while orlistat, C. citrinus extract, and phytosomes were administered at three different concentrations, exhibiting enzyme activities similar to those of the control group. It was also observed that the lowest concentration of phytosomes had a comparable effect to the other concentrations. Callistemon citrinus extract can modulate the activities of enzymes involved in the inflammation process. Furthermore, small doses of phytosomes can serve as anti-inflammatory agents.

Ameliorative Effect of Curcuma longa Ethanolic Extract on the Histology, Hepatic Glycogen Content and Some Biochemical Parameters of the Liver in Streptozotocin-Induced Hyperglycemic Wistar Rats

Introduction: Diabetes mellitus is a life threatening disease that requires immediate medical treatment. Numerous medicinal plants have been known for their anti-diabetic effects and Curcuma longa (turmeric) is one of them. This attribute is possible due to the pharmacological properties of its bio-constituents. In this study, the ameliorative potential of ethanolic extract of Curcuma longa was investigated on the liver of hyperglycemic adult Wistar rats. The objectives were to determine its effects on liver histology, hepatic glycogen content and some liver biochemical parameters. Methods: Forty (40) rats weighing between 120-180g were used for the research. Sixteen (16) rats were used for its acute toxicity study, while twenty-four (24) rats were grouped into four. The groups were labelled A to D and contained six rats each. Group A (normal control) rats were given food pellets and distilled water only. Group B (diabetic control) rats were given 65mg/kg BW of streptozotocin IP. Group C served as the standard group and was given 65mg/kg of streptozotocin IP + 250mg/kg of metformin IP, while Group D (test group) rats were given 65mg/kg BW of streptozotocin IP + 500mg/kg of Curcuma longa extract. Curcuma longa extract was administered through the oral route with the aid of an oral gavage. The administration lasted for twenty-eight (28) days, after which the rats were made to fast overnight with access to only water. The rats were then anaesthetized and whole blood obtained through cardiac puncture for biochemical analysis. Liver tissue was obtained for histological and glycogen content analyses. Results: The results showed severe destruction of hepatocytes with widely spread cytoplasmic vacuolation, loss of hepatic lobules and hepatic inflammation in the diabetic group, whereas it showed prominent nuclei, hepatocytes and distinct sinusoidal spaces in the normal control group. The metformin group showed an intact architecture of the liver tissue with indicating regeneration in the liver, while the Curcuma longa extract group showed a preserved liver cytoarchitecture. There was a significant decrease (p<0.05) in the fasting blood glucose level and liver enzyme activities in the standard and test groups compared to the diabetic group (p<0.05). There was also improvement in glycogen content of the standard and test groups in comparison with the diabetic group (p<0.05). Conclusion: Curcuma longa possesses hepato-protective properties and a strong anti-diabetic potential, therefore it can be considered as an alternative drug in the management of diabetes mellitus and its complications.

In Vivo Study of Physiological, and Histological Effects of Mercury Oxide on Liver and Kidney in Male Wistar Rats

In Vivo Study of Physiological, and Histological Effects of Mercury Oxide on Liver and Kidney in Male Wistar Rats

Pregnancy alters fatty acid metabolism, glucose regulation, and detoxification of the liver in synchrony with biomechanical property changes

Pregnancy places a metabolic burden on the body including the liver, which is responsible for ensuring adequate nutrition for the maternal and fetal systems. To gain a better understanding of liver adaptation, this study investigates metabolic shifts occurring in livers of pregnant rats. Metabolic capacities of the livers of pregnant and non-pregnant female Wistar rats were assessed using comprehensive metabolic models. Kinetic metabolic models were generated for each animal based on protein abundance data from proteomics analysis allowing for a subject-specific assessment of hepatic metabolic functions. Data are available via ProteomeXchange with identifier PXD050758. Additionally, tissue stiffness, viscosity, and water diffusion obtained from magnetic resonance imaging and elastography were correlated with metabolic capabilities to study the relationship between metabolic function and biophysical properties. Proteome profiling revealed differences in protein expression in the livers of pregnant and non-pregnant animals. Functional analysis showed significant variations in metabolic capacities. Livers of pregnant rats had reduced capacities in carbohydrate and fatty acid metabolism, along with altered urea synthesis. Additionally, there were associations between metabolic functions and biophysical properties highlighting potential links between changes in liver structure and metabolic capacities during pregnancy. In summary, our work reveals extensive hepatic metabolic changes in pregnant rats. The liver adapts its metabolic capacities to ensure whole-body metabolic homeostasis but may struggle to counteract nutritional challenges, such as hypoglycemia. The study, employing a personalized approach combining proteomics, kinetic modeling, and advanced imaging, sheds light on the intricate interplay between hepatic adaptations and medical imaging markers, providing a foundation for further investigations into the implications for maternal and fetal health.

Propolis ameliorates renal, liver, and pancreatic lesions in Wistar rats.

This study aimed to evaluate the potential of ethanolic extract of propolis on the secondary lesions of the liver, renal, and pancreatic that were derived by primary colorectal cancer, and comparison of the ethanolic extract of propolis with the vitamin E. The groups included the control, ethanolic extract of propolis, vitamin E, dimethylhydrazine, dimethylhydrazine + ethanolic extract of propolis, and dimethylhydrazine + vitamin E. After 13 weeks of treatment, the blood and tissue samples were taken from all the rats, and alanine transaminase, aspartate transaminase, alkaline phosphatase, uric acid, creatinine, blood urea nitrogen, insulin, amylase, and lipase indices along with the tissue pathological examination of the kidney, liver, and pancreas were evaluated. Ethanolic extract of propolis effectively alleviated the colorectal cancer-induced secondary lesions in the liver by significantly lowering the alanine transaminase significantly. Ethanolic extract of propolis significantly decreased uric acid in rats; and also significantly elevated the pancreatic insulin. In addition, inflammation and cell necrosis indices in all these tissues were significantly reduced when ethanolic extract of propolis was consumed compared to the dimethylhydrazine group. It seemed ethanolic extract of propolis showed high antioxidant, anticancer, and anti-inflammatory potentials, and can be used practically to reduce the side lesions of colorectal cancer.

Uncoupling as initiating event in mitochondrial dysfunction after diuron exposure

Diuron, a herbicide derived from urea, has been shown to induce urinary bladder urothelial tumors in rodents, leading the U.S. Environmental Protection Agency (USEPA) to designate it as a ‘known/likely’ human carcinogen. In our laboratory, a series of studies investigating the carcinogenic mode of action (MoA) of Diuron have consistently revealed its cytotoxic effects on the urinary bladder urothelium. Prolonged exposure to relatively high doses of Diuron results in urothelial necrosis, regenerative hyperplasia, and eventually, the development of tumors. The hypothesis posited is that Diuron and its metabolites exert toxicity by causing damage to mitochondria, a phenomenon referred to as mitotoxicity. Our research focuses on evaluating how Diuron and its metabolites affect mitochondria isolated from both the urothelium and the liver, the primary organ for Diuron biotransformation. In this context, we present and discuss data pertaining to mitochondria isolated from the liver of Wistar rats exposed to Diuron or its metabolites 3-(3,4-diclorofenil)-1-metilureia (DCPMU) or 3,4-dichloroaniline (DCA) at concentrations ranging from 0.5 to 500 µM in vitro. The findings indicate that, at concentrations of 100 and 500 µM, the tested chemicals induce uncoupling of oxidative phosphorylation, as evidenced by the dissipation of mitochondrial membrane potential and basal oxygen consumption. Notably, at 500 µM, DCA causes mitochondrial swelling, a morphofunctional indicator of severe organelle damage. These outcomes underscore the classification of Diuron and its metabolites, DCA and DCPMU, as mitotoxic to liver cells, given the pronounced mitochondrial dysfunction they induce.

Mitigating the effects of time in the heart and liver: The variable effects of short- and long-term caloric restriction

Caloric restriction (CR) is known for its anti-aging benefits, partly due to reduced oxidative stress and enhanced antioxidant defense. However, CR outcomes vary based on its intensity, timing, and duration. This study explored CR's effects on antioxidant activity in the heart and liver of male Wistar rats during aging. We investigated two CR paradigms: long-term CR (LTCR), started early in life, and short-term CR (STCR), initiated in middle or old age for 3 months. Contrary to previous findings of short-term CR deleterious effects of on the nervous system, our results revealed increased levels of key antioxidants after STCR. More specifically, we found an increase in GSH-Px and GSH under STCR that was particularly pronounced in the liver, while an increase in CAT and GR activities was observed in the heart of the STCR groups. Catalase was characterized as an enzyme particularly responsive to CR, as its activity was also increased in both the liver and heart after long-term caloric restriction. Our results highlight a significant tissue-specific response to CR and contribute to our understanding of the dynamic effects of CR, which in turn has implications for refining its therapeutic potential in combating age-related decline.

TOXICITY TEST OF Eusideroxylon zwageri BARK EXTRACT BASED ON LIVER HISTOPATHOLOGY HYDROPIC DEGENERATION AND NECROSIS

Background: Ironwood bark extract contains antioxidant properties such as flavonoids, phenolics, and proanthocyanidins, which can stabilize ROS in the body and help accelerate wound healing. The liver plays a role in nutrient metabolism, detoxification, and ROS production. The lack of antioxidants to neutralize excess ROS makes the liver vulnerable to damage. Purpose: This study aimed to determine that there was no toxic effect of giving ironwood bark extract (Eusideroxylon zwageri) doses of 1.250 mg/kg, 2.750 mg/kg, and 4.750 mg/kg on the liver of Wistar rats based on histopathological features of hydropic degeneration and necrosis. Methods: Pure laboratory experimental quantitative (true experimental) with a post-test-only research design with control group design and One Way Anova statistical test. Treatment was given to 4 groups, namely group (K) was given aquadest control, group (T1) was given ironwood bark extract at a dose of 1.250 mg/kgBW, group (T2) was given ironwood bark extract at a dose of 2.750 mg/kgBW, and group (T3) given ironwood bark extract at a dose of 4.750 mg/kgBW. Results: The results showed no significant difference between the four treatment groups based on the average percentage of hydropic degeneration and necrosis. Conclusion: There was no toxic effect of giving ironwood bark extract doses of 1.250 mg/kgBW, 2.750 mg/kgBW, and 4.750 mg/kgBW on the liver of Wistar rats based on histopathological appearance of hydropic degeneration and necrosis for 14 days.

Chronic aluminium chloride exposure induces redox imbalance, metabolic distress, DNA damage, and histopathologic alterations in Wistar rat liver.

Aluminium, a ubiquitous environmental toxicant, is distinguished for eliciting a broad range of physiological, biochemical, and behavioural alterations in laboratory animals and humans. The present work was conducted to study the functional and structural changes induced by aluminium in rat liver. Twenty five adult male Wistar rats (150-200 g) were randomly divided into five groups; control group and four Al-treated groups viz: Al 1 (25mg AlCl3/kg b.wt), Al 2 (35mg AlCl3/kg b.wt), Al 3 (45mg AlCl3/kg b.wt), and Al 4 (55mg AlCl3/kg b.wt). Rats in the aluminium-treated groups were administered AlCl3 for 30 days through oral gavage. Aluminium significantly increased the serum levels of liver function markers (ALT, AST, and ALP), phospholipids, and cholesterol. The activities of hepatocyte membrane (ALP, GGT, and LAP) and carbohydrate metabolic (G6P, F16BP, HK, LDH, MDH, ME, and G6PDH) enzymes were significantly altered by AlCl3 administration. Prolonged Al exposure induced oxidative stress in the liver, as evident by significant hepatocellular DNA damage, increased lipid peroxidation, and decreased non-enzymatic and enzymatic antioxidants. The toxic effects observed in this study were AlCl3 dose-dependent. Histopathological examination of liver sections revealed enlargement of sinusoidal spaces, derangement of the hepatic chord, loss of discrete hepatic cell boundaries, congestion of hepatic sinusoids, and degeneration of hepatocytes in Al-intoxicated rats. In conclusion, aluminium causes severe hepatotoxicity by inhibiting the hepatocyte membrane enzymes and disrupting the liver's energy metabolism and antioxidant defence.

The Effect of Red Dragon Fruit Juice (Hylocereus Polyrhizus) on Liver Histopathological Images in Wistar Rats Exposed to Cigarette Smoke

Cigarette smoke contains toxic components, carbon monoxide, hydrogen cyanide, carbon dioxide, and tar, which can trigger the formation of ROS (Reactive Oxygen Species) and reduce antioxidant reserves, which impact oxidative stress and are known to cause damage to liver cell membranes. Red dragon fruit (Hylocereus polyrhizus) is a fruit that is rich in antioxidants that inhibit ROS and prevent damage to liver cells by cigarette smoke. This study aims to assess the effect of administering red dragon fruit juice on liver microscopic images in Wistar rats exposed to cigarette smoke. It was an experimental study with a post-test-only control group design. Thirty-five male Wistar rats (Rattus novergicus) were randomized into five groups. Four groups were exposed to cigarette smoke (K-, P1, P2, and P3), and the fifth group was used as a control (K). All the P-groups were given red dragon fruit juice 2 g/2.5 ml, 4 g/2.5 ml, and 8 g/2.5 ml, respectively, for twenty-eight days. On the 29th day, the livers of Wistar rats were taken for preparation and microscopic observation. The research data were analyzed using the Kruskall-Wallis and Mann-Whitney post hoc tests to determine group differences. The average liver cell damage in K, K-, P1, P2, and P3 were 1.23, 2.51, 1.57, 1.51, and 1.43, respectively. There was a significant difference in damage between the K- group and the P1 (p = 0.002), P2 (p = 0.004), and P3 (p = 0.002) groups. The administration of red dragon fruit juice can protect the liver cells of rats exposed to cigarette smoke.

The Protective Activity of Kaempferol Glycosides from Soya Leaf Extract for Experimentally Induced Non-alcoholic Fatty Liver Disease in Wistar Rats

Non-alcoholic fatty liver disease (NAFLD) ranks as one of the most important chronic liver conditions globally, characterized by excessive fat deposition in the liver, central obesity, insulin resistance and in general with characteristics of the metabolic syndrome. Thus, it is vital to treat NAFLD as well as its accompanying metabolic disorders. The present study is to evaluate Kaempferol glycosides activity from Glycine.max leaves extract in experimentally induced NAFLD in rats. The study evaluated the KG activity of G.max extract from leaves on a the HFD and Fructose induced rat model, utilizing Albino Wistar rats with daily doses of 200-250g. Parameters included body weight, liver weight, fasting blood glucose, fasting insulin, insulin resistance, lipid profiles, and histopathological investigations. The doses for KGE were 300mg/kg as medium and 400mg/kg as high. From the research results it was determined that medium dose (300mg/kg) and high dose (400mg/kg) demonstrated a substantial effect when compared to HFD and fructose induced positive control. The study demonstrated that Kaempferol glycosides from Glycine.max leaves extract can prevent and cure HFD and fructose-induced non-alcoholic fatty liver in Albino Wistar rats. The extract's action was substantial at medium and high doses, compared to HFD and fructose-induced positive controls. This shows that Kaempferol glycosides could be a promising natural therapeutic agent against NAFLD, hyperlipidaemic diseases, and metabolic syndromes with less adverse effects compared to conventional medications.

Buchholzia Coriacea Up-Regulates Anti-Oxidant Genes and Represses Inflammatory and Apoptotic Genes in Post Administration of Sub-Chronic Dose of Acetaminophen Induced Hepato-Renal Toxicity in Male Wistar Rats

The aim of this research was to evaluate the possible renal and hepato-protective effects of ethanolic extract of Buchholzia coriacea seed and N-acetylcysteine, on gene expression in the liver and kidneys of Wistar rats subjected to post-treatment with sub-chronic dose of acetaminophen.

Technique of PCR analysis of MT2A genes in different animal species

Metallothioneins are a family of low-molecular-weight cysteine-rich proteins that participate in the metabolism of essential metals and protect against heavy metal toxicity. Metallothioneins are represented by different isoforms in many eukaryotic and prokaryotic species. The promoter regions of metallothionein genes contain sequences of elements that regulate expression in response to metals (MRE), which makes it an urgent task to investigate polymorphisms in the promoter regions of metallothionein genes. Objective. To develop and optimize the technique of PCR analysis of orthologous metallothionein genes in various biological species using the example of the MT2A isoform of rats and pigs. Methods. Analysis of the primary structure of MT2A isoform genes was performed using the NCBI and Ensembl databases. The design ofprimers for PCR analysis was carried out using the Primer3 program. Biomaterial samples were used for the study: livers of Wistar rats and blood of large white pigs of intrabreed type ULW-1. DNA was isolatedfrom rat liver using the NeoPrep DNA Magnet plant DNA kit, and from pig blood using Chelex 100 reagent. Amplification of DNA fragments was carried out using PCR followed by electrophoretic separation of the amplicons in a 2 % agarose gel. Results. In the course of the study, the primary structure of the MT2A genes of rats and pigs was analyzed. Primers were designed to cover the promoter regions of the MT2A genes of these two animal species. The expected sizes of the amplicons are 583 bp and 512 bp for rats andpigs, respectively. The conditions for the synthesis of PCR amplicons were selected, the optimality of which is confirmed by the presence of the corresponding PCR amplicons on the electrophoregram. Conclusions. The developed PCR analysis system for the MT2A genes of rats and pigs allows to study polymorphisms in the promoter regions that contain MRE sites. Considering the importance of MRE sequences for the regulation of metallothioneins’ expression under the influence of metals, as well as the participation of metallothioneins in various physiological and pathological processes, it can be expected that polymorphisms in the studied areas may have associations with adaptation potential, resistance to heavy metals, economic qualities of organisms, etc. In the future, research involving the MT2A gene as a candidate for the development of molecular genetic markers and their use in the practice of marker-associated selection is promising. Keywords: metallothioneins, polymorphism, Rattus norvegicus, Sus scrofa domesticus, polymerase chain reaction, promoter.