Acute exposure of apigenin induces hepatotoxicity in Swiss mice

Apigenin, a dietary flavonoid, is reported to have several therapeutic effects in different diseases including cancer. Toxicity of Apigenin is however, least explored, and reports are scanty in literature. This warrants dose-specific evaluation of toxicity in vivo. In the present study, Apigenin was administered intraperitoneally to Swiss mice at doses of 25, 50, 100 and 200 mg/kg. Serum levels of alanine amino transferase (ALT), aspartate amino transferase (AST) and alkaline phosphatase (ALP) were measured along with the examination of liver histology, reactive oxygen species (ROS) in blood, lipid peroxidation (LPO), glutathione level, superoxide dismutase activity, catalase activity, glutathione S-transferase activity and gene expression in liver tissue. Increase in ALT, AST, ALP, ROS, ratio of oxidized to reduced glutathione (GSSG/GSH) and LPO, altered enzyme activities along with damaged histoarchitecture in the liver of 100 or 200 mg/kg Apigenin treated animals were found. Microarray analysis revealed the differential expression of genes that correspond to different biologically relevant pathways including oxidative stress and apoptosis. In conclusion, these results suggested the oxidative stress induced liver damage which may be due to the regulation of multiple genes by Apigenin at higher doses in Swiss mice.     

The interactive effect of elevated temperature on deltamethrin-induced biochemical stress responses in Channa punctata Bloch

There are reports showing interactive effect of environmental factors with the toxic outcome of chemicals. We studied the interactive effect of elevated temperature as an abiotic stressor on deltamethrin-induced biochemical stress responses in a freshwater fish, Channa punctata Bloch. Heat stress (∼12 °C above ambient temperature for 3 h) and pesticide exposure (deltamethrin 0.75 ppb for 48 h) showed significant induction of heat shock protein-70 (HSP70) in liver, kidney and gills of fishes. Elevated temperature when followed by deltamethrin exposure showed synergistic effect showing a high level of HSP70 in liver and gills whereas response in the kidney was opposite. On the contrary, when deltamethrin exposure followed the heat stress, no significant difference was observed. Protein carbonylation was found to be more pronounced in heat-stressed group compared with control fish group. A significant increase in lipid peroxidation (LPO) was observed in different tissues of fish exposed to either of the stressors. In the kidney of fish exposed to heat stress followed by deltamethrin, LPO was relatively lower as compared to other treatments. Thiols content such as reduced glutathione (GSH), total thiols (T-SH), non-protein thiols (NP-SH) and protein thiols (P-SH) showed no consistent pattern in different tissues. In deltamethrin-exposed group that was subsequently exposed to heat stress, the GSH content was higher in liver and lower in both kidney and gills when compared with other groups. Alteration in the activities of antioxidant enzymes such as catalase (CAT), glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx) was also observed when fish were exposed to heat stress and/or deltamethrin. Our study demonstrated that heat stress modulated biochemical stress responses in fish showing a tissue specific pattern. This implies that fish has the capacity to elicit differential response to exposure to abiotic stressors in order to reduce the systemic magnitude of stress which may otherwise lead to severe dysfunction of vital tissues.     

Fenvalerate induced biochemical changes in the selected tissues of freshwater fish, Cyprinus carpio

Alterations in the levels of some biochemical and enzymological parameters in brain, liver and muscle tissues of Cyprinus carpio exposed to sublethal concentration (10 micrograms/liter) of fenvalerate for 6, 12, 24 and 48 hr were studied. Total, structural and soluble proteins were decreased, whereas the free amino acids and the activities of protease, aspartate aminotransferase and alanine aminotransferase significantly increased in fenvalerate exposed fish. It is also observed that the changes were more pronounced with an increase in the period of exposure. Within the tissues, the alterations in biochemical and enzymological parameters was in the following order: liver greater than brain greater than muscle. The possible roles of fenvalerate induced changes in the fish are discussed.     

The activity of pro- and antioxidant systems in the liver of freshwater fishes during different seasons

The content of lipid peroxidation products--diene conjugates, lipid hydroperoxides, thiobarbituric acid reactive substances (TBARS), vitamins A, E and carotenoids and the activity of antioxidant enzymes--superoxide dismutase, glutathione peroxidase and catalase in the liver of freshwater fishes of different species (silver carp, grass carp and common carp) in different seasons have been studied. It was established the activity of antioxidant defence system in the liver of fish depends significantly on the season and fish species. In particular, the content of lipid peroxidation products in the liver of freshwater fishes at the beginning of winter and spring was significantly higher compared to their content at the beginning of summer and autumn. The superoxide dismutase and glutathione peroxidase activities in the liver of these fish species at the beginning of winter and spring were significantly lower than at the beginning of summer and autumn while the seasonal changes of catalase activity in the liver of fish are expressed insignificantly. The content of vitamins E, A1, A2 and carotenoids in the liver of fishes of different species at the beginning of winter and spring was significantly lower than at the beginning of summer and autumn. The content of lipid peroxidation products and vitamins E, A1 and A2 in the liver of common carp is significantly lower than in the liver of silver carp and grass carp and species differences in antioxidant enzymes activity are insignificant.     

Effect of bisphenol A toxicity on growth performance,biochemical variables,and oxidative stress biomarkers of Nile tilapia,Oreochromis niloticus (L.)

Bisphenol A (BPA) is one of the industrial chemical compound which is used in the production of polycarbonate plastics and epoxy resins. BPA is used throughout the world and it could enter the aquatic ecosystems causing serious problems. To evaluate the potential effects of BPA toxicity on Nile tilapia, Oreochromis niloticus (L.) performance, its lethal concentration (LC₅₀) was determined and it was 13.13 µg/L. After that, fish (33.9 ± 0.55 g/fish) were exposed to 0.0, 1.64, or 3.28 µg/L of BPA for 6 weeks after which growth performance, biochemical variables, and oxidative defense system were assessed. The results showed that fish growth and feed intake were significantly reduced as BPA levels increased with no significant difference in fish survival. Total protein, albumin, globulin, and acetylcholine esterase decreased significantly; meanwhile, aspartate transferase, alanine transferase, alkaline phosphatase, uric acid, and creatinine increased significantly with exposure to BPA in a dose dependent manner. Furthermore, malondialdehyde value and the activities of superoxide dismutase and catalase increased significantly; while glutathione peroxidase and glutathione S‐transferase decreased significantly as BPA levels increased. In conclusion, BPA exposure in aquatic environment deteriorated fish performance and health causing liver and kidney dysfunction. Thus, fish exerted oxidative defense enzymes as a protection tool against BPA toxicity.     

Effects of cadmium toxicity upon the in vivo and in vitro activity of proteins and five enzymes in blood serum and tissue homogenates of Mugil cephalus

This study reports changes in total protein and certain liver, heart, gill and serum enzymes after exposing fish or tissue homogenates and serum to Cd, i.e. in vivo and in vitro effects. Five enzymes were selected for assay; aspartate amino transferase (AAT), alanine amino transferase (A1AT), alkaline phosphatase (A1P), acid phosphatase (ACP) and lactic dehydrogenase (LDH). Total protein content in the exposed fish showed an increase in the liver, gill and serum while there was no change in heart protein. The sensitivity of the five assayed enzymes to Cd varied in different tissues. Gill AAT, A1AT were the most sensitive. Liver A1P and heart LDH showed the maximum inhibition at higher Cd concentrations.     

Toxic and sub-lethal effects of oleandrin on biochemical parameters of fresh water air breathing murrel, Channa punctatus (Bloch.)

Active compound oleandrin extracted from Nerium indicum (Lal Kaner) leaf has potent piscicidal activity. The piscicidal activity of oleandrin on freshwater fish C. punctatus was both time and dose dependent. Exposure to sub-lethal doses of oleandrin for 24hr and 96hr to fish caused significant alteration in the level of total protein, total free amino acid, nucleic acid, glycogen, pyruvate, lactate and enzyme protease, phosphatases, alanine aminotransferase, aspartate aminotransferase and acetylcholinesterase activity in liver and muscle tissues. The alterations in all the above biochemical parameters were also significantly time and dose dependent. The results show a significant recovery in all the above biochemical parameters, in both liver and muscle tissues of fish after the 7th day of the withdrawal of treatment. Toxicity persistence test of oleandrin on juvenile Labeo rohita shows that fish seed of common culturing carp can be released into rearing ponds after three days of oleandrin treatment. It supports the view that the oleandrin is safer and may be useful substitute of other piscicides for removing the unwanted freshwater fishes from aquaculture ponds.     

Alterations in the general condition,biochemical parameters and locomotor activity in Cnesterodon decemmaculatus exposed to commercial formulations of chlorpyrifos,glyphosate and their mixtures

The Pampean region, an extensive area of South America is continuously impacted by agricultural activities and the pesticides related to them like chlorpyrifos and glyphosate. Both pesticides have been registered in freshwater bodies of the region. One of the most abundant and widely distributed fish species in Pampean streams is Cnesterodon decemmaculatus, which have to cope with this altered scenario.In the present study the toxicity of Clorfox^® and Roundup Max^®, the commercial formulations of chlorpyrifos and glyphosate, respectively, and their mixture where evaluated using a set of biomarkers at different biological organization levels in fish exposed to relevant environmentally pesticides concentrations. Somatic indexes such as the condition factor (K), and the hepato-somatic index (HSI), the locomotor activity through the distance traveled and the average speed, the enzymatic activities of acetylcholinesterase (AChE) in brain and muscle, catalase (CAT) in muscle and liver, glutathione-S-transferase (GST) in brain, liver, muscle and gills, aspartate amino-transferase (AST), alanine amino-transferase (ALT), AST/ALT ratio and alkaline phosphatase (ALP) in liver were measured on C. decemmaculatus. Adult females were exposed during 6 weeks to the following concentrations: 0.0084 μl/l and 0.00084 μl/l of Clorfox (CF), 0.2 and 2 mg/l of Roundup Max (RM) and all the combinations of these concentrations. The CF exposure caused a decrease in the condition factor and in the locomotor activity parameters and induced an increase brain AChE, liver CAT activity and AST/ALT ratio. On the other hand, the exposure to RM produced a decrease in liver GST, AST/ALT ratio and ALP activity. Finally, some pesticide combinations decrease general condition and liver GST activities, and increase brain GST and liver ALP activities. Different responses in biomarkers were observed in mixtures treatments, reflecting the complex interactions between these toxics and suggesting a suppressive action of RM on CF effects.Since the concentrations we tested are environmentally relevant and the overall fish health condition was affected, the presence of these pesticides in freshwater systems could impose a risk for populations by causing deleterious effects on C. decemmaculatus in Pampean region.     

Protective effect of black tea extract on the levels of lipid peroxidation and antioxidant enzymes in liver of mice with pesticide-induced liver injury

Sub-acute hepatotoxicity was induced in mice by exposure to pesticides. The effect of pretreatment with aqueous black tea extract on lipid peroxidation and antioxidants in the liver was investigated. Administering a combination dose of chlorpyriphos and cypermethrin (20 mg kg(-1) each) on alternate days over a 15-day period to male mice resulted in induction of sub-acute toxicity as reflected by elevated levels of liver damage marker enzymes alkaline phosphatase(ALP), aspartate transaminase(AST) and alanine transaminase(ALT). Significantly elevated levels of lipid peroxidation were observed in the experimental group (group III) as compared with control mice. Decreased activities of superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), total thiol, glutathione peroxidase (GPx), glutathione reductase(GR) and glutathione-S-transferase (GST) were also observed in pesticide-treated as compared to control mice. Aqueous black tea extract was given as a pretreatment to group IV mice at a dose of 200 mg ml(-1) polyphenols before the pesticide dose, which significantly decreased the levels of lipid peroxidation and significantly elevated the activities of SOD, CAT, GSH, total thiol, GPx, GR and GST in liver to levels similar to the controls. Thus, the data offer support for the claim that the central mechanism of pesticide action occurs via changes in cellular oxidative status and shows conclusively that supplementation with black tea extract protects against the free radical-mediated oxidative stress in hepatocytes of animals with pesticide-induced liver injury.     

Expression of some hepatocyte-like functional properties of WRL-68 cells in culture

Summary Some morphologic and functional characteristics of an hepatic fetal human epithelial cell line (WRL-68 cells) were determined to validate the use of these cells as an in vitro hepatic model. WRL-68 cells have a morphologic structure similar to hepatocytes and hepatic primary cultures. They secrete alpha-feto protein and albumin and exhibit a cytokeratin pattern similar to other hepatic cultures. WRL-68 cells preserve the activity of some characteristic or specific liver enzymes or both used in clinical chemistry for the diagnosis of hepatic disorders, i.e. alanine amino transferase, aspartate amino transferase, gamma-glutamyl transpeptidase, and alkaline phosphatase.     

Sublethal effects of an organophosphorus insecticide (RPR-II) on biochemical parameters of tilapia, Oreochromis mossambicus

The effect of exposure to sublethal concentrations (0.017 mg L(-1), 1/10 of LC50) of the novel organophosphate (OP) insecticide, 2-butenoic acid-3-(diethoxyphosphinothioyl) methyl ester (RPR-II) on biochemical parameters in Oreochromis mossambicus was studied during exposure for 3, 7, 15, 30 and its recovery response after seven days. Acetylcholinesterase (AChE) activity of brain, gill and muscle was inhibited by 67%, 77% and 73% respectively on day-30. The plasma and kidney alanine aminotransferase (AlaAT), and aspartate aminotransferase (AspAT) activity increased, while decreases were observed in gill and liver. Increases in acid phosphatase (AcP), and alkaline phosphatase (AP) activities were observed in plasma, gill, and kidney, and reductions of 20% and 61% in liver AcP and AP, respectively. Depletion of glycogen was observed in all tissues, an indication of typical stress related response of the fish with pesticide. Lactate dehydrogenase (LDH) activity decreased in liver and muscle, indicating tissue damage but a significant increase in LDH activity in gill and brain was observed. Depletion of glutathione (GSH) was observed in all tissues, thereby enhancing lipid peroxidation resulting in cell damage. The induction in hepatic glutathione-S-transferase (GST) levels indicates protection against the toxicity of xenobiotic-induced lipid peroxidation. There was a significant recovery in the above biochemical parameters, in all tissues of fish after a recovery period of seven days. These results revealed that the OP insecticide RPR-II is highly toxic and affects the intermediary metabolism of O. mossambicus.     

In vivo impact of monocrotophos on biochemical parameters of a freshwater fish during subacute toxicity and following cessation of exposure to the insecticide

In vivo toxicity of monocrotophos on key metabolites and enzymes of the protein metabolism was investigated in important tissues of the freshwater fish Clarias batrachus. Fish were exposed to 1/10 and 1/20 of LC50 concentration for 28 days. After 28 days of exposure, some fish were transferred to monocrotophos-free water and kept in the same for 21 days (recovery period) in order to study the recovery response. Total protein, amino acid, and ammonia contents were decreased in gill, kidney, liver, and muscle tissues, and recovery was slight at the end of 21 days of transfer of fish into freshwater. Urea and glutamine levels were elevated, except in kidneys, and recovered at the end of the recovery period. The activities of protease, transaminase, and phosphatase enzymes were elevated in all tissues during 28 days of exposure and at both concentrations. Recovery of the activity of enzymes was more significant at the lower concentration as compared to the higher concentration.     

Role of berberine in ameliorating Schistosoma mansoni-induced hepatic injury in mice

Background

Schistosomiasis is caused by helminth parasites of the genus Schistosoma. Berberine chloride (BER), an isoquinoline alkaloid, has been used in vivo for its antiparasitic, antioxidant and hepatoprotective properties. In this study, the protective effect of BER and praziquantel has been compared for the extent of schistosomiasis-induced oxidative stress in hepatic tissue of mice.

Results

S. mansoni was able to induce inflammation and injury to the liver, evidenced (i) by an increase in inflammatory cellular infiltrations, dilated sinusoids and vacuolated hepatocytes, (ii) by decreased levels of alanine and aspartate aminotransferases and increased levels of alkaline phosphatase, γ-glutamyl transferase in the liver homogenate, (iii) by increased production of nitric oxide and thiobarbituric acid reactive substances, and (iv) by lowered glutathione levels and decreased activities of catalase and superoxide dismutase, respectively. All these infection-induced parameters were significantly altered during BER treatment. In particular, berberine counteracted the S. mansoni-induced loss of glutathione and the activities of catalase and superoxide dismutase.

Conclusion

Based on these results, it is concluded that berberine could ameliorate pre-existing liver damage and oxidative stress conditions due to schistosomiasis.     

Lipid peroxidation induced by Clinostomum detruncatum in muscle of the freshwater fish Rhamdia quelen

The effect of Clinostomum detruncatum metacercaria infection on the activities of the antioxidant enzymes superoxide dismutase and catalase in muscle of the freshwater fish Rhamdia quelen was analyzed. Tert-butyl hydroperoxide-initiated chemiluminescence, a measure of lipid peroxidation, was also investigated. Enzyme activities were similar in infected and uninfected fishes. However, the chemiluminescence was almost 2-fold higher in muscle of infected fishes than in muscle of uninfected ones. These results indicate that parasite infection induces oxidative stress and a higher level of membrane damage in the fish muscle due to an imbalance between pro-oxidants and non-enzymatic antioxidants. Our results suggest that fish response to parasite infection could involve, as in other vertebrates, reactive oxygen intermediates.     

Antioxidant and hepatoprotective activity of Aphanizomenon flos-aquae Linn against paracetamol intoxication in rats

Paracetamol caused liver damage as evident by significant increase in the activities of aspartate and alanine transferases. There were general statistically significant losses in the activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione transferase and an increase in thiobarbituric acid reactive substances in the liver of paracetamol treated group compared with the control group. However, treatment with ethanol extract of A. flos-aquae (EEAFA) was able to counteract these effects. Protection offered by silymarin (standard reference drug) seemed relatively greater. The results suggest that EEAFA can act as hepatoprotective agent against paracetamol induced toxicity as an antioxidant.     

Abrogation by Trifolium alexandrinum root extract on hepatotoxicity induced by acetaminophen in rats

Abstract

Objective

Acetaminophen (APAP) is a substance that harms human health by stimulating free radical production. This study investigated the ability of Trifolium alexandrinum root (TAR) extract to reduce the hepatotoxicity induced by APAP in rats.

Methods

Animals were classified into four groups and treated for 6 weeks. Group 1: normal control-treated (saline); Group 2: TAR extract-treated (100 mg/kg); Group 3: APAP-treated; Group 4: APAP plus TAR extract.

Results

APAP significantly elevated AST (aspartate amino transferase), ALT (amino alanine transferase), ALP (alkaline phosphatase), GGTP (gamma glutamyl transpeptidase), bilirubin, and malondialdehyde with a significant decrease in glutathione, superoxide dismutase, glutathione peroxidase, catalase, and glutathione S-transferase compared with the control group. Administration of TAR extract combined with APAP improved the liver damage induced by APAP. Histopathological evidence, together with observed DNA fragmentation, supported the detrimental effect of APAP and the ameliorating effect of TAR extract on liver toxicity.

Conclusion

TAR extract has beneficial properties and can reduce the liver damage and toxicity induced by APAP.

Discussion

Free radical mediated processes have been implicated in the pathogenesis of many diseases. The protective effect of TAR root extract on APAP-induced hepatotoxicity in rats appears to be related to inhibition of lipid peroxidation and enhancement of antioxidant enzyme levels, in addition to a free radical scavenging action.     

Amelioration of tamoxifen-induced liver injury in rats by grape seed extract, black seed extract and curcumin

Liver injury was induced in female rats using tamoxifen (TAM). Grape seeds (Vitis vinifera) extract (GSE), black seed (Nigella sativa) extract (NSE), curcumin (CUR) or silymarin (SYL) were orally administered to TAM-intoxicated rats. Liver histopathology of TAM-intoxicated:rats showed pathological changes. TAM-intoxication elicited declines in liver antioxidant enzymes levels (glutathione peroxidase, glutathione reductase, superoxide dismutase and catalase), reduced glutathione (GSH) and GSH/GSSG ratio plus the hepatic elevations in lipid peroxides, oxidized glutathione (GSSG), tumor necrosis factor-alpha (TNF-alpha) and serum liver enzymes; alanine transaminase, aspartate transaminase, alkaline phosphatase, lactate dehydrogenase and gamma glutamyl transferase levels. Oral intake of NSE, GSE, CUR or SYL to TAM-intoxicated rats, attenuated histopathological changes and corrected all parameters mentioned above. Improvements were prominent in case of NSE (similarly SYL) > CUR > GSE. Data indicated that NSE, GSE or CUR act as free radicals scavengers and protect TAM-induced liver injury in rats.     

The effects of selenium on oxidative stress biomarkers in the freshwater characid fish matrinxã, Brycon cephalus (Günther, 1869) exposed to organophosphate insecticide Folisuper 600 BR (methyl parathion)

Methyl parathion (MP), an organophosphate widely applied in agriculture and aquaculture, induces oxidative stress due to free radical generation and changes in the antioxidant defense system. The antioxidant roles of selenium (Se) were evaluated in Brycon cephalus exposed to 2 mg L(-1) of Folisuper 600 BR (MP commercial formulation - MPc, 600 g L(-1)) for 96 h. Catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione S-transferase (GST), reduced glutathione (GSH) and lipid peroxidation (LPO) levels in the gills, white muscle and liver were evaluated in fish fed on diets containing 0 or 1.5 mg Se kg(-1) for 8 weeks. In fish treated with a Se-free diet, the MPc exposure increased SOD and CAT activities in all tissues. However, the GPx activity decreased in white muscle and gills whereas no alterations were observed in the liver. MPc also increased GST activity in all tissues with a concurrent decrease in GSH levels. LPO values increased in white muscle and gills and did not change in liver after MPc exposure. A Se-supplemented diet reversed these findings, preventing increases in LPO levels and concurrent decreases in GPx activity in gills and white muscle. Similarly, GSH levels were maintained in all tissue after MPc exposure. These results suggest that dietary Se supplementation protects cells against MPc-induced oxidative stress.     

Biochemical evaluation of hepatic damage in subchronic exposure to malathion in rats: effect on superoxide dismutase and catalase activities using native PAGE

The aim of this study was the evaluation of the hepatic damages following a subchronic exposure to malathion, an organophosphorus (OP) insecticide. Malathion was administered intragastrically in 1 ml corn oil containing 100 mg/kg Body Weight daily for 32 days. Malondialdehyde (MDA) concentration superoxide dismutase (SOD) and catalase (CAT) activities were analysed using a non-denaturing electrophoresis. The serum activities of Pseudocholinesterase (PchE), aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) were determined. Malathion exposure leads to a significant decrease in AchE activity, an increase in hepatic MDA, and in serum ASAT and ALAT activities. A positive correlation between serum transaminases levels and hepatic MDA was demonstrated. These results indicate that malathion exposure induced lipid peroxidation LPO, a process of degradation of membrane lipids, involving the deterioration of the cellular integrity. We have recorded a slight increase in antioxidant enzymes activities. This leads us to suggest an insufficient elimination of free radicals, causing cytotoxic effects. To cite this article: R. Rezg et al., C. R. Biologies 331 (2008).