Effect of alcohol and kolanut interaction on brain sodium pump activity in Wistar Albino rats.

Effect of alcohol-kolanut interaction on Sodium Pump activity in wistar albino rats was studied. Thirty wistar albino rats were divided into six groups of five (5) rats per group and used for the study. The control group (1) received via oral route a placebo (4ml of distilled water). Groups 2 to 6 were treated for a period of 21 days, with (10% v/v) of alcohol (group 2), 50mg/kg body weight of kolanut (group 3), 50 mg/kg body weight of caffeine (group 4), 4ml of 10% v/v of alcohol and 50 mg/kg body weight kolanut (group 5), 4ml of 10% v/v of alcohol and 50 mg/kg body weight of caffeine in 4.0ml of the vehicle via gastric intubation respectively. A day after the final exposure, the brain of each rat was harvested and processed to examine several biochemical parameters, i.e., total ATpase, ouabain-insensitive ATpase, ouabain sensitive ATpase (Na(+)-K(+)ATPase), non-enzymatic breakdown of ATP and inorganic phosphate (Pi) released. The results showed that the essential enzyme of the brain responsible for neuronal function, Na(+)-K(+)ATPase, was inhibited by alcohol-kolanut co-administration relative to control, resulting in a decrease in Na(+)-K(+)ATPase activity, ATP production, ion transport and action potential, leading to loss of neuronal activities.     

Effect of caffeine-coconut products interactions on induction of microsomal drug-metabolizing enzymes in Wistar Albino rats.

Effect of caffeine-coconut products interactions on induction of drug-metabolizing enzyme in wistar albino rats was studied. Twenty rats were randomly divided into four groups: The control group (1) received via oral route a placebo (4.0ml of distilled water). Groups 2 to 4 were treated for a 14-day period with 50 mg/kg body weight of caffeine, 50 mg/kg body weight of caffeine and 50 mg/kg body weight of coconut water, and 50 mg/kg body weight of caffeine and 50 mg/kg body weight of coconut milk in 4.0ml of the vehicle via gastric intubation respectively. One day after the final exposure, the animals were anaestheticized by inhalation of an overdose of chloroform. The blood of each rat was collected by cardiac puncture while the liver of each rat was harvested and processed to examine several biochemical parameters, i.e., total protein and RNA levels, protein/RNA ratios, and activities of alanine and aspartate amino transferase (ALT and AST, respectively). The results showed that while ingestion of coconut milk and coconut water increased the values of protein and protein/RNA ratios, it decreased alanine and aspartate amino transferase (ALT and AST) activities. These effects, in turn, enhanced the induction of the metabolizing enzymes and a resultant faster clearance and elimination of the caffeine from the body, there by reducing the toxic effect on the liver.     

Trichosanthes dioica Fruit Ameliorates Experimentally Induced Arsenic Toxicity in Male Albino Rats Through the Alleviation of Oxidative Stress

The present work was focused to evaluate the ameliorative property of aqueous extract of Trichosanthes dioica fruit (AQ T. dioica fruit) against arsenic-induced toxicity in male Wistar albino rats. AQ T. dioica fruit was administered orally to rats at 50 and 100 mg/kg body weight for 20 consecutive days prior to oral administration of sodium arsenite (10 mg/kg) for 10 days. Then the rats were sacrificed for the evaluation of body weights, organ weights, hematological profile, serum biochemical profile, and hepatic and renal antioxidative parameters viz. lipid peroxidation, reduced and oxidized glutathione, glutathione-S-transferase, glutathione peroxidase, glutathione reductase, superoxide dismutase, catalase, and DNA fragmentation. Pretreatment with AQ T. dioica fruit at both doses markedly and significantly normalized body weights, organ weights, hematological profiles, and serum biochemical profile in arsenic-treated animals. Further, AQ T. dioica fruit pretreatment significantly modulated all the aforesaid hepatic and renal biochemical perturbations and reduced DNA fragmentation in arsenic-intoxicated rats. Therefore, from the present findings, it can be concluded that T. dioica fruit possessed remarkable value in amelioration of arsenic-induced hepatic and renal toxicity, mediated by alleviation of arsenic-induced oxidative stress by multiple mechanisms in male albino rats.     

Cellular and biochemical aspects of growth retardation in rat fetuses induced by maternal administration of selected anticancer agents.

Single ip injections of 600 mg/kg 5-(3,3-dimethyl-1-triazeno)-imidazole-4-carboxamide (DIC) and 900 mg/kg 5-[3,3-bis(2-chlorethyl)-1-triazeno]-imidazole-4-carboxamide (BIC) were given to pregnant Wistar rats at day 12 and the animals were killed 4 h after injection and at days 13-17 of gestation. Fetal tissues were used to determine total DNA, RNA, and protein and the data used to derive cell number and cell weight, RNA, and protein/cell. Both compounds reduced total fetal body weight, DNA, RNA, and protein but reduction of RNA by BIC was not statistically significant. These effects were observed 4 h after injection, increased with age (days 13-17), and were 3-4 times greater for DIC than BIC. By using the value of 6.2 mumug DNA/cell, cell number and per-cell values for weight, RNA, and protein, and weight: DNA, RNA:DNA, and protein:DNA ratios were computed. The per-cell values and ratios in the DIC-exposed animals were 8-44% greater and in BIC-treated animals 0-11% greater than control animals of the same gestational age. Percentage of body water was the same in the experimental and control animals. The differences in DNA, RNA, and protein are believed to be related to drug-induced growth retardation incident to total fetal DNA reduction resulting in diminished cell number.     

Early embryonic development in the rat following in utero exposure to alcohol and caffeine

The influence of both alcohol and caffeine on early embryonic development was investigated in pregnant rats. Compared to the corresponding controls, a high incidence of resorptions and abnormal embryos was induced following treatment of the animals with alcohol (0.015 ml/g body weight, 12.5% v/v, i.p.) on gestational days 6 through 12 and with caffeine (25 mg/kg body weight, i.v.) on gestational day 10. In addition, embryonic growth was severely affected. Reduction of placental blood circulation and impairment of cellular proliferation may account for the observed deleterious effects on the embryo.     

Effects of caffeine and bombesin on ethanol and food intake

The methylxanthine caffeine and ethyl alcohol are widely used and powerful psychotropic drugs, but their interactions are not well understood. Bombesin is a brain-gut neuropeptide which is thought to function as a neurochemical factor in the inhibitory control of voluntary alcohol ingestion. We assessed the effects of combinations of intraperitoneal (i.p.) doses of caffeine (CAF, 0.1-50 mg/kg) and bombesin (BBS, 1-10 micrograms/kg) on 5% w/v ethanol solution and food intake in deprived rats. Deprived male and female Wistar rats received access to 5% ethanol or Purina chow for 30 minutes after i.p. injections. In single doses, CAF and BBS significantly decreased both ethanol and food consumption, at 50 mg/kg and 10 micrograms/kg, respectively. CAF and BBS combinations produced infra-additive, or less-than-expected inhibitory effects on ethanol intake, but simple additive inhibitory effects on food intake. This experimental evidence suggests a reciprocal blocking of effects of CAF and BBS on ethanol intake but not food intake. Caffeine, when interacting with bombesin, increases alcohol consumption beyond expected values. Caffeine could affect the operation of endogenous satiety signals for alcohol consumption.     

Homeostasis changes induced by the action of ethanol on the materno-fetal complex in rats. IV. Late maternal effects following acute intoxication during the preimplantation period

The possible late maternal effect on biochemical homeostasis of acute administration of ethanol during the preimplantation period was studied on pregnant albino female rats (Wistar strain). Females were injected i.v. with a 33.16% (v/v) solution of ethanol (4.80 ml/kg body weight) on day 2 and 4 of pregnancy, the animals being sacrificed on day 20. The effects on hepatic DNA biosynthesis and on some serum metabolites (proteins, lipids and some non-protein nitrogen compounds) were determined in the treated and in an untreated control group. In the treated group a non-significant increase of hepatic DNA concentration was found. Homeostasis changes of serum proteins involved: a slight increase of total serum protein content, hypoalbuminemia and hyperglobulinemia (non-significant values). Lipid metabolites showed also non-significant changes: increase of total lipids and decrease of cholesterol. Uric acid and urea (non-protein metabolites) concentration was increased. This increase, in spite of its significance level, must be taken into account and may be due to the presence of dead fetuses and to the consecutive renal lesions.     

Boron ameliorates arsenic‐induced DNA damage,proinflammatory cytokine gene expressions,oxidant/antioxidant status,and biochemical parameters in rats

Arsenic, an element found in nature, causes hazardous effects on living organisms. Meanwhile, natural compounds exhibit protective effects against hazardous substances. This study evaluated the effects of boron against arsenic‐induced genotoxicity and altered biochemical parameters in rats. Thirty‐five male Wistar albino rats were equally divided into five groups, and the experimental period lasted 30 days. One group was used as the control, and another group was treated with 100 mg/L arsenic in drinking water. The other groups were orally treated with 5, 10, and 20 mg/kg boron plus arsenic (100 mg/L via drinking water). Arsenic caused changes in biochemical parameters, total oxidant/antioxidant status, and DNA damage in mononuclear leukocytes. Moreover, it increased IFN‐γ, IL‐1β, TNF‐α, and NFκB mRNA expression levels in rat tissue. However, boron treatment improved arsenic‐induced alterations in biochemical parameters and increases in DNA damage and proinflammatory cytokine gene expressions.     

Modulatory role of lipoic acid on adriamycin-induced testicular injury

The present study investigated the protective efficacy of dl-alpha-lipoic acid (LA) on adriamycin (ADR)-induced oxidative damage in rat testis. Adult male albino rats of Wistar strain were administered ADR (1 mg/kg body weight, i.v.), once a week for 10 weeks. ADR injected rats showed increased oxidative stress with a concomitant decrease in cellular thiols. The mRNA level for phospholipid hydroperoxide glutathione peroxidase (PHGPx) was also significantly decreased by ADR administration. Transmission electron microscopic (TEM) observations of testicular germ cells revealed abnormal ultrastructural changes in ADR treated rats. Treatment with lipoic acid (35 mg/kg body weight, i.p.) 1 day prior to ADR administration, effectively reverted these abnormal changes towards normalcy. These findings indicate a cytoprotective role of LA in this experimental model of testicular toxicity.     

Protein-energy malnutrition during pregnancy alters caffeine's effect on brain tissue of neonate rats

We studied whether protein-energy malnutrition changed brain susceptibility to a small dose of caffeine in newborn rats. Since we had demonstrated previously that caffeine intake during lactation increased the brain neuropeptide on newborns, we investigated further the effects of the prenatal administration of caffeine on TRH and cyclo (His-Pro). From day 13 of gestation to delivery day, pregnant rats in one group were fed either a 20% or a 6% protein diet ad libitum, and those in the other group were pair-fed with each protein diet supplemented with caffeine at an effective dose of 2 mg/100 g body weight. Upon delivery, brain weight, brain protein, RNA, DNA and the neuropeptides thyrotropin-releasing hormone (TRH) and cyclo (His-Pro) were measured in the newborn rats. A 6% protein without caffeine diet caused reductions in brain weights and brain protein, RNA and DNA contents, but did not alter brain TRH and cyclo (His-Pro) concentrations in the newborn animals. In the offspring from dams fed a 6% protein diet, caffeine administration significantly elevated brain weights and brain contents of protein, RNA and DNA. In contrast, these values were similar between noncaffeine and caffeine-supplemented animals in a 20% protein diet group. Brain TRH and cyclo (His-Pro) concentrations were not changed by caffeine administration. These data suggest that caffeine augments protein synthesis in the newborn rat brain when malnourished, but that the same dose of caffeine did not affect protein synthesis in brains of newborn rats from normally nourished dams. Therefore, the present findings indicate that the nutritional status of mothers during pregnancy has important implication in the impact of caffeine on their offspring's brains.     

Lung mechanics and connective tissue proteins in diabetic Bio-Breeding/Worcester Wistar rats

We studied lungs of spontaneously diabetic Bio-Breeding/Worcester (BB/W) Wistar rats which resemble human insulin-dependent diabetes mellitus. Compared with the age-matched control group, the body weight of the diabetic rats tended to be smaller and lung wet and dry weight were similar, but lung dry weight, relative to body weight and to lung wet weight, was significantly larger. Both air and saline lung volumes were reduced in the diabetic rats, and volume-pressure (V-P) curves expressed as a percent of maximal lung volume were significantly shifted downward and to the right of those in the control group over the midportion. Total DNA and RNA contents were similar in both groups, whereas protein content and concentration and protein/DNA and RNA/DNA ratios were significantly reduced in the diabetic rats. In contrast, content and concentration of 4-hydroxy-L-proline, elastin, and crude connective tissue were significantly higher in the diabetic group. We conclude that the increase in connective tissue proteins in the BB/W rats is most likely responsible for the shift in the V-P curves.     

Toxicological evaluation of karaya gum; acute and subacute oral toxicity in rats

Male and female albino rats (Wistar strain) were given single and multiple doses of karaya gum suspended either in peanut oil or mixed with basal diet at different concentrations ranging from 0.5 to 8 g gum/kg body weight. The plant gum did not elicit any overt signs of toxicity or death in both sexes of rats. Daily administration of karaya gum mixed with basal diet at different dose levels (0, 5, 20 and 40 g gum/kg diet) for a period of 90 days showed no adverse effects in male and female rats. The body weight, growth pattern, food and water intake were comparable with those of the normal rats. There were no significant biochemical, or morphological alterations in the vital organs of experimental animals.     

Correlation of histological an histometric changes in rats testes treated with chloroquine phosphate.

Histological and histometric changes in the testes of albino Wistar rats were correlated. Wistar rats weighing between 180-240g were randomly divided into three groups of ten rats each. One group served as control and the rats were given normal saline. The second and third groups received 2mg/kg and 4mg/kg body weights of chloroquine phosphate daily for thirty days respectively. Seminiferous tubules of animals treated with chloroquine phosphate were irregular in shape and were also isolated compared to control. Marked disruption of the inter-tubular stroma of testes in the treated groups was also observed. Histometric variations in testicular tissue was observed in the experimental animals following treatment with chloroquine phosphate. The 2mg/kg body weight and 4mg/kg body weight animals recorded significantly lower [P< 0.05] relative germinal epithelial volume of 43.95 % and 32.70 % respectively when compared to the control (51.75 %). The volume of stroma in the third group (49.33 %) was significantly higher [P < 0.05] when compared to the control (16.83 %) and 2mg/kg body weight rats (22.83 %). We observed negative correlation coefficient between lumen and seminiferous tubular volume in the control group compared to the other groups which showed a positive correlation. Correlation between germinal epithelium and seminiferous tubular volume were positive in all groups. These findings have thrown more light on recognized histological changes by accurately grading these changes which offers objectivity and increased precision compared with direct visual appraisal.     

Role of alpha-tocopherol in cadmium-induced oxidative stress in Wistar rat's blood, liver and brain

Cadmium (Cd) a highly toxic metal is considered to be a multitarget toxicant, and it accumulates principally in the liver and kidney after absorption. In vivo studies of mouse and rat liver have shown that apoptosis plays a primary role in Cd-induced hepatotoxicity. However, the detailed mechanisms by which toxic metals such as Cd produce their effects are still largely unknown. The present study aimed at investigating the consequences of exposure to Cd, alpha-tocopherol and their combination on stress biochemical parameters (lipoperoxidation and protein carbonyls levels). Male albino Wistar rats (1 month old) were treated intravenously with cadmium (2 mg CdCl(2)/kg body weight/day), and alpha-tocopherol (100 mg/kg body weight/day), or with alpha-tocopherol+Cd (100 mg Vit E/kg body weight, 2 mg CdCl(2)/kg). The lipoperoxidation was measured by the thiobarbituric acid reactive substances (TBARS) method and oxidatively generated damage to proteins by determining carbonyl (DNPH) levels. Among the hematological parameters measured the haematocrit value and haemoglobin concentration were significantly decreased in the blood of Cd-treated rats. A significant increase was observed in the level of malondialdehyde (MDA) and protein carbonyls in the cadmium exposed group compared to control group (p<0.001), and these values were decreased after administration of alpha-tocopherol (group 4). The activity of lactate dehydrogenase in rat liver and brain showed a significant increase as compared to that found in the control group and significant decrease of catalase and superoxide dismutase activities. In the liver of the Cd-treated group the contents of reduced glutathione were decreased. Our results suggest that cadmium induces an oxidation of cellular lipids and proteins and that administration of alpha-tocopherol can reduce Cd-induced oxidative stress and improve the glutathione level together with other biochemical parameters.     

Protective effect of betaine on changes in the levels of lysosomal enzyme activities in heart tissue in isoprenaline-induced myocardial infarction in Wistar rats

Myocardial infarction is one of the most common manifestations of cardiovascular disease. In the present study, we investigated the protective effect of betaine, a potent lipotropic molecule, on changes in the levels of lysosomal enzymes and lipid peroxidation in isoprenaline-induced myocardial infarction in Wistar rats, an animal model of myocardial infarction in man. Male albino Wistar rats were pretreated with betaine (250 mg/kg body weight) daily for a period of 30 days. After the treatment period, isoprenaline (11 mg/100 g body weight) was intraperitoneally administered to rats at intervals of 24 h for 2 days. The activities of lysosomal enzymes (β-glucuronidase, β-galactosidase, β-glucosidase, and acid phosphatase) were significantly (p < 0.05) increased in plasma with a concomitant decline in the activities of these enzymes in heart tissue of isoprenaline-administered rats. Also, the level of lipid peroxidation was higher in heart lysosomes of isoprenaline-injected rats. Pretreatment with betaine daily for a period of 30 days to isoprenaline-induced rats prevented the changes in the activities of these lysosomal enzymes. Oral treatment with betaine (250 mg/kg body weight) to normal control rats did not show any significant effect in all the biochemical parameters studied. Thus, the results of our study show that betaine protects the lysosomal membrane against isoprenaline-induced myocardial infarction. The observed effects might be due to the free radical-scavenging and membrane-stabilizing properties of betaine.     

Effect of nickel on testicular nucleic acid concentrations of rats on protein restriction

The nucleic acids (DNA and RNA) and total protein concentration in testes were estimated in male Wistar strain rats treated intraperitorally with nickel sulfate (2.0 mg/100 g body weight) on alternate days for 10 dosages. In both normal (18% casein) and protein-restricted (5% casein) experimental animals, the nucleic acids and total protein concentration were found to decrease significantly compared to the corresponding controls. Sperm count and sperm motility were also reduced in both experimental groups of animals. The results indicate that nickel influences the expression of genetic information by reducing testicular nucleic acids and protein concentration in both dietary experimental groups.     

Remedial effect of DL-α-lipoic acid against adriamycin induced testicular lipid peroxidation

Adriamycin (ADR), a cytotoxic antineoplastic drug is used in the treatment of various solid tumors. However, its efficacy continues to be challenged by significant toxicities including testicular toxicity. In the present study, the effect of lipoic acid, a "universal antioxidant" was investigated on ADR induced peroxidative damages in rat testis. Adult male albino rats of Wistar strain were administered ADR (1 mg/kg body weight, i.v.) once a week for 10 weeks. ADR injected rats showed a significant decline in the activities of enzymic antioxidants (catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase and glutathione-S-transferase) and non-enzymic antioxidants (reduced glutathione, Vitamin A, Vitamin C and Vitamin E) with high malondialdehyde levels. The extent of testicular toxicity was evident from the decreased activities of testicular marker enzymes (sorbitol dehydrogenase and glucose-6-phosphate dehydrogenase). Treatment with lipoic acid (35 mg/kg body weight, i.p.) one day prior to ADR administration, maintained near normal activities of the enzymes and significantly reduced lipid peroxidation, thereby proving it to be an effective cytoprotectant.     

Caffeine suppresses the expression of the Bcl-2 mRNA in BeWo cell culture and rat placenta

Chronic caffeine exposure during pregnancy has an effect on fetal growth; however, the adverse effects of caffeine on embryogenesis are not well understood and controversial. We used cDNA microarray technology to determine whether caffeine alters gene expressions in a human cytotrophoblast-like cell line, BeWo. We found that the expression of the B-cell CLL/lymphoma 2 (Bcl-2) gene in BeWo cells was down-regulated by caffeine, suggesting that chronic exposure during the gestational period could exert an influence on embryogenesis. We then focused on the Bcl-2- and Bcl-2-associated X protein gene, Bax, to study the responsive gene expression in BeWo cells as well as placentas of pregnant rats fed a diet supplemented with caffeine (2 mg/100 g body weight) during gestation, and analyzed the gene expressions using LightCycler-based quantitative real-time polymerase chain reaction assays. We found a significantly decreased level of Bcl-2 mRNA expression, which demonstrated the influence of caffeine on placental function.     

The effect of caffeine in the in vivo SCE and micronucleus mutagenicity tests

Caffeine which was administered per os to outbred mice either twice, 30 and 6 h before sacrifice or once, 30 h before sacrifice, at dose levels of 50, 75 or 100 mg/kg body weight only caused a weak induction of micronuclei at the highest dose. Again a level of 100 mg caffeine per kg body weight was required before a weak but not significant effect could be observed in the micronucleus test using a mutagen-sensitive inbred strain of mice. In Chinese hamsters caffeine doses of 45, 75, 150 or 300 mg/kg body weight either given once or twice per os at the same time schedule as used for the mice also caused a clear cut induction of micronuclei only at the highest dose level. In the SCE test with Chinese hamster again 300 mg of caffeine were necessary to obtain a mutagenic effect although this test is considered to be more sensitive to mutagenic damage than the micronucleus test. It can therefore be concluded that caffeine causes DNA damage only at dose levels in the LD50 range which is higher for hamsters than for mice.     

The role of phosphoenolpyruvate carboxykinase in neuronal steroidogenesis under acute inflammation

Phosphoenolpyruvate carboxykinase (PEPCK) is a key gluconeogenic enzyme found in many tissues throughout the body including brain. In the present study, we have investigated the effect of bacterial lipopolysaccharide (LPS) on PEPCK and its role in neuronal steroidogenesis. Adult female albino rats were administered LPS (5 mg/kg body weight) to induce acute inflammation. LPS administration resulted in a significant increase of PEPCK mRNA expression with concomitant increase in mRNA levels of steroidogenic acute regulatory (StAR) protein and other steroidogenic enzymes including 3β-hydroxysteroid dehydrogenase (3β-HSD), 17β-hydroxysteroid dehydrogenase (17β-HSD) and aromatase in brain tissue. Further, the inhibition of PEPCK expression by glipizide significantly decreased the mRNA expression of steroidogenic proteins and concurrently increased the mRNA levels of proinflammatory cytokines under LPS administration. The results of this study suggest a novel finding that PEPCK may have an important role in neuronal steroidogenesis; which serves as an adaptive response under inflammation.