Toxic copper level increases erythrocyte glycolytic rate, glutathione production and alters electrolyte balance in male Wistar rats.

BACKGROUND: Copper is a micronutrient vital to several cellular energy metabolic processes and drives erythropoiesis. However, it disrupts cellular biological activities and causes oxidative damage when in excess of cellular needs. This study investigated the effects of copper toxicity on erythrocyte energy metabolism in male Wistar rats. METHODS: Ten Wistar rats (150-170 g) were randomly divided into 2 groups: control (given 0.1 ml distilled water) and copper toxic (given 100 mg/kg copper sulphate). Rats were orally treated for 30 days. Blood, collected retro-orbitally after sodium thiopentone anaesthesia (50 mg/kg i.p.) into fluoride oxalate and EDTA bottles, was subjected to blood lactate assay and extraction of red blood cell respectively. Red blood cell nitric oxide (RBC NO), glutathione (RBC GSH), adenosine triphosphate (RBC ATP) levels, RBC hexokinase, glucose-6-phosphate (RBC G6P), glucose-6-phosphate dehydrogenase (RBC G6PDH), and lactate dehydrogenase (RBC LDH) activity was estimated spectrophotometrically. Values (Mean±SEM, n = 5) were compared by Student's unpaired T-test at p < 0.05. RESULTS AND CONCLUSION: Copper toxicity significantly increased RBC hexokinase (23.41 ± 2.80 µM), G6P (0.48 ± 0.03 µM), G6PDH (71.03 ± 4.76nmol/min/ml) activities, ATP (624.70 ± 57.36 µmol/gHb) and GSH (3.08 ± 0.37 µM) level compared to control (15.28 ± 1.37 µM, 0.35 ± 0.02 µM, 330.30 ± 49.58 µmol/gHb, 54.41 ± 3.01nmol/min/ml and 2.05 ± 0.14 µM respectively, p < 0.05). Also, RBC LDH activity (145.00 ± 19.88mU/ml), NO (3.45 ± 0.25 µM) and blood lactate (31.64 ± 0.91 mg/dl) level were lowered significantly compared to control (467.90 ± 94.23mU/ml, 4.48 ± 0.18 µM and 36.12 ± 1.06 mg/dl respectively). This study shows that copper toxicity increases erythrocyte glycolytic rate and glutathione production. This increase could be connected to a compensatory mechanism for cellular hypoxia and increased free radical generation.

Amelioration of thyroid dysfunction by magnesium in experimental diabetes may also prevent diabetes-induced renal impairment.

BACKGROUND: Diabetes mellitus has been reported to cause thyroid dysfunction, which may also impair renal function. Magnesium has been reported to exert ameliorative effects in diabetes mellitus. This study investigated thyroid and renal functions in experimental type-2-diabetic Wistar rats. METHODS: Experimental type-2-diabetes was induced using short duration high-fat (30%) diet feeding followed by single-dose streptozotocin (35 mg/kg i.p.). Fifty rats were randomly divided into five equal groups consisting of control, diabetes untreated, diabetes treated with either magnesium (250 mg/kg) or metformin (250 mg/kg) and diabetes treated with both metformin and magnesium simultaneously.All treatments were carried out orally for 14days post-diabetes induction. Body weight and blood glucose was monitored using the tail tipping method before diabetes induction and thereafter on days 1,7,14 post-treatment respectively. Thereafter, blood samples were collected by cardiac puncture after light anesthesia into plain and EDTA sample bottles. Total protein, albumin, globulin (plasma) and insulin (serum) were assayed in all samples obtained. Thyroid stimulating hormone (TSH), triiodothyronine, thyroxine was also evaluated (n = 5/group) in serum while blood urea nitrogen (BUN), creatinine was assessed (n = 5/group) in plasma. Kidney homogenates were obtained per group and analyzed for renal superoxide dismutase (SOD), reduced glutathione (GSH) and lipid peroxidation (MDA). Kidney histology was also evaluated per group using both Haematoxylin and Eosin and periodic acid Schiff stains. RESULTS: Body weight, blood glucose, insulin, renal MDA was increased in diabetic untreated compared to other groups. Reductions (P < 0.05) in TSH, triiodothynine, Renal SOD and GSH levels where observed in diabetic untreated compared to other groups. Renal histology in diabetic untreated showed glomerula sclerosis, fused messengial cells and either collapsed tubular lumen or lumen with eosinophilic renal cast. These pathologies where partially reversed in the other experimental groups. CONCLUSION: This study suggests that thyroid and renal impairment may be present in experimental type-2-diabetes. Treatment with oral magnesium may cause a partial restoration of thyroid function that may impede the development of renal dysfunction.

Pathophysiology of iron overload-induced renal injury and dysfunction: Roles of renal oxidative stress and systemic inflammatory mediators.

Iron-overload has been recognized as a risk factor for organ dysfunction and damage resulting in diseases such as liver and heart disease, diabetes mellitus, and neurodegenerative diseases. This study investigated renal function and some systemic inflammatory indices in iron-overloaded male Wistar rats. Thirty animals were equally distributed into 3groups and treated daily i.p. with either normal saline (0.2 ml; control), iron (as ferrous sulphate) (15 mg/kg) or iron (30 mg/kg) for 21days respectively. Post-treatment, blood samples were obtained from each animal by cardiac puncture after light anaesthesia into plain sample bottles. Iron, ferritin, transferrin, creatinine, urea, albumin, total protein, interleukin-6 (IL-6), prostaglandins-E2 and tumor necrosis factor-α (TNF-α) were analysed in serum. Kidney homogenates were obtained per group and analysed for superoxide dismutase (SOD), total antioxidant capacity (TAC), reduced glutathione (GSH), lipid peroxidation (MDA) and nitric oxide (NO). Kidney histology was evaluated per group using both Haematoxylin and Eosin and periodic acid Schiff stains. Iron-overload caused a graded increase (p < 0.05) in serum iron, ferritin, transferrin, creatinine, urea, IL-6, TNF-α, TAC, MDA and NO levels as well as a reduction in albumin levels, renal SOD and GSH in groups 2 (iron 15 mg/kg) and 3 (iron 30 mg/kg) respectively compared to control. Histological evaluation of the kidney showed structural and tubular aberrations consistent with renal damage via inflammatory processes in iron overloaded rats. Our present study suggests that iron-overloading causes renal dysfunction by triggering the evolution of several inflammatory mediators which lead to a cascade of systemic and renal inflammatory processes that alter renal structure and function.

Effect of methanol extract of musa sapientum leaves on protein glycation and erythrocyte antioxidant status in alloxan-induced diabetic Wistar rats.

BACKGROUND: Increased generation of free radicals from protein glycation has been associated with compromised integrity of erythrocytes in diabetes. Musa sapientum has been reported to possess anti-diabetic properties and this study investigated the effect of methanol extract of Musa sapientum on protein glycation and erythrocyte integrity. METHODS: Forty-two male Wistar rats (180-200g) were randomly grouped into seven: 1 (control), 2 (diabetic untreated), 3 (normal extract-treated (250 mg/kg)), 4 (normal metformin-treated (150 mg/kg)), 5 (diabetic extract-treated (250 mg/kg)), 6 (diabetic metformin-treated (150 mg/kg)), 7 (diabetic insulin-treated (1 IU/kg)). Diabetes was induced with single intraperitoneal injection of 120 mg/kg alloxan. Animals were treated for 14 days and blood (3 mls) was collected from retro-orbital plexus to determine serum fructosamine level, erythrocyte superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities. Glycated haemoglobin (HbA(1c)) level was estimated using a conversion formula. Animals were sacrificed thereafter by cervical dislocation and pancreatic tissues were excised and stained with haematoxylin and eosin for histological study. Statistical significance at P < 0.05 was analyzed by one-way ANOVA and Newman-Keuls' post-hoc test. RESULTS: Diabetic rats treated with extract, metformin and insulin had significant reduction in serum fructosamine level by 62.64%, 74.63% and 56.05% respectively while HbA(1c) level reduced by 45.06%, 50.62% and 40.57% respectively. Activities of erythrocyte SOD and GPx were increased in the extract-treated group. Histological studies showed regeneration of islet cells in the diabetic extract-treated rat which was comparable to normal. CONCLUSION: The extract inhibited protein glycation, regenerated the islet cells and improved erythrocyte antioxidant status in diabetic rats.