Bromobenzene Treatment Research Articles

Protective effect of Cassia fistula fruit extract against bromobenzene-induced liver injury in mice

In the present study, hepatoprotective effect of Cassia fistula fruit extract was investigated in mice. Animals were divided into six groups receiving normal saline (1), bromobenzene (460 mg/kg) alone (2) and together with increasing doses (200, 400, 600, 800 mg/kg) of a crude hydro-alcoholic extract of Cassia fistula fruit (3-6, respectively). All administrations were carried out orally, daily, for 10 days. On the 11th day, animals were sacrificed. Serum activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and gamma glutamyl transpeptidase (γGT) were determined; serum levels of direct and total bilirubin were measured; furthermore, livers were prepared for histological examination. Our results showed that bromobenzene treatment alone elicited a significant increase in activities of AST, ALT, ALP (but not γGT), and it significantly elevated the levels of direct and total bilirubin. Co-treatment with Cassia fistula fruit extract, however, significantly and dose-dependently decreased the above-mentioned enzyme activities (with exception of γGT) and bilirubin levels, producing a recovery to the naive state. The protective effect of Cassia fistula fruit extract against liver injury evoked by bromobenzene was confirmed by histological examination as well. In conclusion, the Cassia fistula fruit extract has significant hepatoprotective effect in our murine model.

Protective effect of ginger extract against bromobenzene-induced hepatotoxicity in male rats

The bromobenzene (BB)-induced hepatotoxicity comes from its reactive metabolites. The efficacy of different doses of ginger ( Zingiber officinales Rose) extract in alleviating hepatotoxicity was investigated in male albino rats. Oxidative stress parameters were monitored. The drugs metabolizing enzymes; cytochrome P450 and GST, pro-inflammatory marker; COX-2 and the apoptotic marker; caspase-3 were assessed. Animals were assigned to 1 of 5 groups: control group; bromobenzene (460 mg/kg BW) alone, three animal groups 3–5 treated with different doses of ethanolic ginger extract (100, 200, 300 mg/kg BW, respectively) 2 weeks prior bromobenzene (460 mg/kg BW) treatment. Rats received orally ginger extract daily for 21 days whereas bromobenzene treatment for 7 days starting from 15th day of treatment. Oral treatment of BB was found to elicit a significant decrease in the activities of the antioxidant enzymes; SOD, GPx and the GSH level, while the activities of GR and drug metabolizing enzymes; GSTs and Cyt P450 were enhanced. Also, BB-treatment resulted in a great enhanced production of nitric oxide products and activation of COX-2 and caspase-3. Pre-treatment with different doses of ginger extract prior to BB-treatment alleviated its toxic effects on the tested parameters in the three animal groups.

IDENTIFICATION OF GLUTATHIONE DEPLETION-RESPONSIVE GENES USING PHORONE-TREATED RAT LIVER

To identify candidate biomarker gene sets to evaluate the potential risk of chemical-induced glutathione depletion in livers, we conducted microarray analysis on rat livers administered with phorone (40, 120 and 400 mg/kg), a prototypical glutathione depletor. Hepatic glutathione content was measured and glutathione depletion-responsive gene probe sets (GSH probe sets) were identified using Affymetrix Rat Genome 230 2.0 GeneChip by the following procedure. First, probe sets, whose signal values were inversely correlated with hepatic glutathione content throughout the experimental period, were statistically identified. Next, probe sets, whose average signal values were greater than 1.5-fold compared to those of controls 3 hr after phorone treatment, were selected. Finally, probe sets without unique Entrez Gene ID were removed, ending up with 161 probe sets in total. The usefulness of the identified GSH probe sets was verified by a toxicogenomics database. It was shown that signal profiles of the GSH probe sets in rats treated with bromobenzene were strongly altered compared with other chemicals. Focusing on bromobenzene, time-course profiles of hepatic glutathione content and gene expression revealed that the change in gene expression profile was marked after the bromobenzene treatment, whereas hepatic glutathione content had recovered after initial acute depletion, suggesting that the gene expression profile did not reflect the hepatic glutathione content itself, but rather reflects a perturbation of glutathione homeostasis. The identified GSH probe sets would be useful for detecting glutathione-depleting risk of chemicals from microarray data.

Acquired Resistance to Bromobenzene Hepatotoxicity by Repeated Treatment of Rats with Bromobenzene

It has often been observed that chemical-induced initial insult is no longer detected in examinations after additional consecutive treatments, suggesting that the target organs acquire resistance to chemical toxicity. In this study, we examined whether acquired resistance to toxicity would be observed after repeated treatments of a toxic dose of bromobenzene (BB). In Experiment 1, F344 male rats were intraperitoneally given BB at a dosage of 150 mg/kg/day. Based on the serum AST level 20 h after the first treatment, the rats were divided into 2 groups, Groups 2 and 3, which consisted of rats with elevated AST and rats without remarkable changes in AST activity, respectively. Subsequently, the rats were subjected to the same BB treatment regimen for 4 or 9 consecutive days. The control group (Group 1) was administered with the vehicle. The AST activity of Group 2 showed no remarkable changes from Day 3, indicating an acquired resistance to BB hepatotoxicity. Measurements of drug-metabolizing enzymes in Group 2 demonstrated a reduction in CYP contents and activities, and a strong induction of GST, which contributed to the resistance. The aminotransferase activities in Group 3, however, showed no changes throughout the dosing periods. In Experiment 2, the rats with the same BB treatment regimen as Experiment 1 were administered intraperitoneally with a single dosage of BB of 300 mg/kg. Although a higher dosing of BB caused hepatic injury in all three groups, the degree of injury in the two groups with BB treatment was much slighter than that in the control (vehicle + BB). These results indicate that not only Group 2 but also Group 3 acquired resistance to BB hepatotoxicity after repeated treatments. The exposure levels of BB in the two groups were lower than those of the control. Thus, changes in the drug-metabolizing reaction related to the metabolism and detoxification of BB contributed to the resistance to BB hepatotoxicity.

Antimutagenic activity of flavonoids from the heartwood of Rhus verniciflua

Pretreatment of the methanolic extract of the heartwood of Rhus verniciflua (Anacardiaceae) to rats prevented the activation of hepatic microsomal cytochrome P 450 enzymes, inhibition of hepatic glutathione S-transferase by bromobenzene treatment, respectively, and therefore significantly decreased malondialdehyde content in the rat. The Ames test showed that the addition of 1.0 mg/plate of the methanolic extract or the EtOAc fraction of the Rhus verniciflua heartwood extract potentially inhibited the mutagenicity by aflatoxin B 1. Column chromatography of the EtOAc fraction yielded four flavonoids, garbanzol ( 1), sulfuretin ( 2), fisetin ( 3), fustin ( 4), mollisacasidin ( 5). When these components were subjected to the Ames test, it was found that sulfuretin might effectively prevent the metabolic activation or scavenge electrophilic intermediates capable of causing mutation. In contrast, fustin showed a dose-independent antimutagenic activity and it has mutagenic/antimutagenic activity. However, a mixture of sulfuretin and fustin (1:1) exhibited dose-dependent antimutagenicity indicating that sulfuretin inhibited the mutagenicity of fustin. These results suggest that the extract of Rhus verniciflua heartwood containing flavonoid complex could be a potent anticarcinogen.

Toxicogenomics of bromobenzene hepatotoxicity: a combined transcriptomics and proteomics approach

Toxicogenomics is a novel approach integrating the expression analysis of thousands of genes (transcriptomics) or proteins (proteomics) with classical methods in toxicology. Effects at the molecular level are related to pathophysiological changes of the organisms, enabling detailed comparison of mechanisms and early detection and prediction of toxicity. This report addresses the value of the combined use of transcriptomics and proteomics technologies in toxicology. Acute hepatotoxicity was induced in rats by bromobenzene administration resulting in depleted glutathione levels and reduced average body weights, 24 hr after dosage. These physiological symptoms coincided with many changes of hepatic mRNA and protein content. Gene induction confirmed involvement of glutathione- S-transferase isozymes and epoxide hydrolase in bromobenzene metabolism and identified many genes possibly relevant in bromobenzene toxicity. Observed glutathione depletion coincided with induction of the key enzyme in glutathione biosynthesis, γ-glutamylcysteine synthetase. Oxidative stress was apparent from strong upregulation of heme oxygenase, peroxiredoxin 1 and other genes. Bromobenzene-induced protein degradation was suggested from two-dimensional gel electrophoresis, upregulated mRNA levels for proteasome subunits and lysosomal cathepsin L, whereas also genes were upregulated with a role in protein synthesis. Both protein and gene expression profiles from treated rats were clearly distinct from controls as shown by principal component analysis, and several proteins found to significantly change upon bromobenzene treatment were identified by mass spectrometry. A modest overlap in results from proteomics and transcriptomics was found. This work indicates that transcriptomics and proteomics technologies are complementary to each other and provide new possibilities in molecular toxicology.

Inhibition of Glutathione Synthesis with Propargylglycine Enhances N-Acetylmethionine Protection and Methylation in Bromobenzene-Treated Syrian Hamsters

The finding that liver necrosis caused by the environmental glutathione (GSH)-depleting chemical, bromobenzene (BB) is associated with marked impairment in O- and S-methylation of BB metabolites in Syrian hamsters raises questions concerning the role of methyl deficiency in BB toxicity. N-Acetylmethionine (NAM) has proven to be an effective antidote against BB toxicity when given after liver GSH has been depleted extensively. The mechanism of protection by NAM may occur via a replacement of methyl donor and/or via an increase of GSH synthesis. If replacement of the methyl donor is an important process, then blocking the resynthesis of GSH in the methyl-repleted hamsters should not decrease NAM protection. This hypothesis was examined in this study. Propargylglycine (PPG), an irreversible inhibitor of cystathionase, was used to inhibit the utilization of NAM for GSH resynthesis. Two groups of hamsters were pretreated with an intraperitoneal (ip) dose of PPG (30 mg/kg) or saline 24 h before BB administration (800 mg/kg, ip). At 5 h after BB treatment, an ip dose of NAM (1200 mg/kg) was given. Light microscopic examinations of liver sections obtained 24 h after BB treatment indicated that NAM provided better protection (P < 0.05) in the PPG + BB + NAM group than in the BB + NAM group. Liver GSH content, however, was lower in the PPG + BB + NAM group than in the BB + NAM group. The Syrian hamster has a limited capability to N-deacetylated NAM. The substitution of NAM with methionine (Met; 450 mg/kg) resulted in a higher level of GSH in the BB + Met group than in the BB + NAM group (P < 0.05). The enhanced protection by PPG in the PPG + BB + NAM group was accompanied by higher (P < 0.05) urinary excretions of specificO- and S-methylated bromothiocatechols than in the BB + NAM group. The results suggest that NAM protection occurs primarily via a replacement of the methyl donor and that methyl deficiency occurring in response to GSH repletion plays a potential role in BB toxicity.

Possible modulatory effect of browned yam flour diet on chemically-induced toxicity in the rat

The possible modulatory effect of browned yam flour, a local dietary staple in south western Nigeria, on the toxicity of 7,12-dimethylbenzanthracene (DMBA), 3-methylcholanthrene (3-MC), carbon tetrachloride (CCl 4) and bromobenzene (BrB) in rats was investigated. Feeding rats with 25% browned yam flour 2 wk before treatment with 65 mg/kg DMBA (single dose) and 5 mg/kg 3-MC and continued for 3 wk significantly decreased the reduction in final body weight or weight gain and organ weights caused by the two compounds. Similarly, the diet decreased the reduction in body weight or weight gain and the increase in relative liver weight mediated by oral treatment with 0.5 ml CCl 4/kg and 2.5 mmol BrB/kg body weight. Incorporation of 25% browned yam flour into rat diet significantly reduced the DMBA-mediated decrease in haemoglobin content, packed cell volume, red blood cell count and white blood cell count by 7, 5, 20 and 10%, respectively; while the diet reduced the 3-MC-mediated decrease in these parameters by 15, 28, 9 and 17%, respectively. The same diet elicited 23, 45, 13 and 33% decreases in CCl 4 mediated reduction in these parameters and 23, 18, 16 and 29% in the case of BrB. Browned yam flour diet caused 10, 14 ( P < 0.001) and 4% ( P < 0.05) reductions in the DMBA-mediated increase in serum aspartate aminotransferase, alanine aminotransferase and serum alkaline phosphatase, respectively; and 32, 31 ( P < 0.05) and 13% ( P < 0.001) in the case of the 3-MC-mediated increase. Also, the diet reduced CCl 4-mediated increase in the activities of these by 40, 34 and 31%, respectively and by 23, 30 and 29% following BrB treatment. These results suggest that browned yam flour diet could possibly be a modulator of chemically induced toxicity.

Induction of hsp 70 in HepG2 cells in response to hepatotoxicants

The objective of this study was to determine if a variety ofhepatotoxicants could induce the level of heat shock protein 701, and whether or not elevated levels of heat shock proteins (hsp's) could provide cytoprotection from those hepatotoxicants. Exposure of HepG2 cells to cytotoxic concentrations of bromobenzene, cadmium, cyclophosphamide, or diethylnitrosamine increased the level of hsp 70I protein and mRNA, while carbon tetrachloride and cocaine had no effect on hsp 70I or mRNA levels. To determine if induction of hsp 70I might afford protection against cytotoxicity, HepG2 cells were given a prior sublethal heat shock (sub-LHS) (43†C for 1 hr) to induce hsp's and then challenged 24 hr later with the hepatotoxicants. Sub-LHS pretreatment diminished toxicity from bromobenzene, cadmium, cyclophosphamide, or diethylnitrosamine, but not carbon tetrachloride or cocaine. In cells treated with [14C]carbon tetrachloride or [3H]cocaine, no detectable covalent biinding to proteins was observed; whereas, [14C]-bromobenzene treatment resulted in substantial covalent binding to cellular protein. The apparent absence of formation of reactive metabolite adducted proteins from cocaine and carbon tetrachloride may explaiin why no hsp 70I induction was observed with these agents. The correlation between hepatotoxicant induction of hsp 70I and cytoprotection afforded by sub-LHS pretreatment suggests that hsp 70I induction may represent an important cellular defense mechanism in the liver.

In vivo and in vitro 31P magnetic resonance spectroscopic studies of the hepatic response of healthy rats and rats with acute hepatic damage to fructose loading.

The hepatic response to a fructose challenge for control rats, and rats subjected to an acute sublethal dose of carbon tetrachloride (CCl4) or bromobenzene (BB), was compared using dynamic in vivo 31P MRS. Fructose loading conditions were used in which control rats showed only a modest increase in hepatic phosphomonoester (PME), and a small decrease in ATP, Pi, and intracellular pH after fructose administration. Both CCl4 and BB-treated rats showed a much greater fructose-induced accumulation of PME than did controls. Trolox C, a free radical scavenger, prevented most of this PME increase. BB-treated rats, given sufficient time to recover from the hepatotoxic insult, responded to the fructose load similarly to controls. Liver aldolase activities of control, toxicant-treated rats, and toxicant plus Trolox C-treated rats correlated inversely with PME accumulation after fructose loading (correlation coefficient: -0.834, P < 0.05). Perchloric acid extracts of rat livers studied by in vitro 31P MRS confirmed that the PME accumulation after fructose loading is mainly due to an increase in fructose 1-phosphate. These studies are consistent with the aldolase-catalyzed cleavage of fructose 1-phosphate being rate-limiting in hepatic fructose metabolism, and that the CCl4 and BB treatment modify and inactivate the aldolase enzyme.

Bromobenzene accelerates hepatocyte streaming in rats

Sixty male adult rats weighing between 250 and 300 g were divided in two equal experimental groups. The first experiment was designed to evaluate hepatocyte streaming. Animals were injected with 0.5 μCi [3H]thymidine and 1 hour later received one intraperitoneal injection of bromobenzene (3.8 mmol/kg, dissolved in corn oil). The animals were then killed in groups of five at 1 hour and 2,4,7,14, and 30 days. The aim of the second experiment was to evaluate labeling index changes with time. Animals received one intraperitoneal injection of bromobenzene and were killed in groups of five at 1, 2, 3, 4, 7, and 14 days. They received [3H]thymidine 1 hour before killing. Bromobenzene induced a necrosis in the third acinus zone that disappeared within a week. On day 0 the labeling index of hepatocytes and littoral cells was 0.3% ± 0.04% and 0.4% ± 0.05%, respectively. On the second day, it reached 14.6% ± 2.6% and 10.1% ± 3.1% and returned to its initial value after 1 week. Dead cells in the third zone were replaced by inflowing cells from the intact zones. Hepatocytes and littoral cells streamed at the same velocity of 6 ± 0.5 μm/day, faster than in untreated animals with velocity of 3.2 μm/day. Parenchyma and stroma responded to injury in a coordinated fashion. From the functional point of view, hepatocytes and littoral cells operate as a unit that is called proliferon. The maximal proliferon life span was 57 days after bromobenzene treatment and 108 days in controls.

Potentiation of bromobenzene-induced pneumotoxicity by phenobarbital as determined by bronchoalveolar lavage fluid analysis.

Bromobenzene produces pulmonary, renal and hepatic damage in the rat. Phenobarbital potentiates bromobenzene-induced hepatotoxicity. Studies were initiated to determine if phenobarbital potentiates the pulmonary damage produced by bromobenzene. In a dose ranging study, adult male rats were treated daily for 4 days with phenobarbital (80 mg/kg, ip) and on the fifth day were dosed with 0, 2, 3, or 4 mmoles/kg, ip, bromobenzene. In a larger study phenobarbital was given for 4 days (80 mg/kg, ip), and bromobenzene (3.3 mmoles/kg, ip) was given on the fifth day. Pulmonary damage was assessed 24 hours later in the first study and 12 hours later in the second study by bronchoalveolar lavage fluid analysis (BALF), microsomal mixed function oxidase measurements and histopathological evaluations. BALF biochemical markers employed were gamma-glutamyl transpeptidase (GGT), lactate dehydrogenase (LDH) and total protein. Phenobarbital treatment greatly enhanced bromobenzene-induced GGT and LDH release into the lavage fluid in a dose-dependent manner. Phenobarbital treatment increased microsomal 7-O-dealkylation of both ethoxy- and pentoxyresorufin in the liver without affecting these activities in the lung. Bromobenzene treatment decreased the hepatic microsomal dealkylation of decreased the hepatic microsomal dealkylation of pentoxyresorufin in a dose-dependent manner. Since known rat pulmonary P450 isozymes have been reported to be insensitive to phenobarbital induction, it may be that the toxicity is due to transport of a reactive metabolite(s) formed in the liver to the lung or bromobenzene is activated by some other pulmonary P450 isozyme responsive to phenobarbital.

The role of acinar zone 3 hepatocytes in bile formation: influence of bromobenzene treatment on bile formation in the rat

The role of zone 3 hepatocytes in bile formation was determined when they were selectively destroyed by 3.8 mmol/kg b.w. of bromobenzene injected i.p. for 48 h, as compared to appropriate controls. Bromobenzene treatment resulted in 29 +/- 4.4% hepatic lobule necrosis localized in the zone 3 hepatocytes. Although bile flow and bile salt-independent flow were not affected, this treatment was associated with a significant reduction in bile salt, and phospholipid secretion. The bile salt pool and bile salt synthesis were also significantly decreased. These results suggest that necrosis of zone 3 hepatocytes induced by bromobenzene reduced bile acid synthesis which decreased bile salt pool and affected bile salt and phospholipid secretion rates. However, necrosis of zone 3 hepatocytes did not affect bile flow or the bile salt-independent flow, suggesting that hepatocytes of zones 1 and 2 maintained the normal bile salt-independent flow when zone 3 hepatocytes were damaged.

BROMOBENZENE-INDUCED CLARA CELL DAMAGE: THE CONTRIBUTION OF CYTOCHROME P-450 SYSTEM LOCALIZED IN THE CLARA CELL

The relationship between the alterations of different pulmonary drug metabolizing systems and the selective damages of Clara cells in ddY mice caused by bromobenzene treatment was investigated in order to elucidate the mechanism of bromobenzene-induced pneumotoxicity. Pulmonary drug metabolizing systems showed different responses to oral administration of bromobenzene (5 mmole/kg body weight) in mice at 8 and 24 hours. The time-dependent decrease of coumarin hydroxylase activity was the severest compared to other drug metabolizing enzymes. Bromobenzene treatment caused selective damages of the Clara cells without influencing the ciliated bronchiolar cells and alveolar type II cells, and the degrees of Clara cell damage became severer with time. These results suggest that the degradation of pulmonary coumarin hydroxylase is closely related to the selective damage of Clara cells induced by bromobenzene, and this enzyme contributes to the occurrence of bromobenzene-induced Clara cell damages.

Microheterogeneity of biliverdin reductase in rat liver and spleen: Selective suppression of enzyme variants in liver by bromobenzene

Recently we have reported the detection of multiple net-charge and molecular mass variants of biliverdin reductase in the rat liver. We now report an apparent selective change in the electrophoretic profile of the reductase variants in the liver by in vivo bromobenzene treatment (2 mmol/kg, sc, 24 h). Using two-dimensional electrophoresis and isoelectric focusing, one molecular mass species of the reductase ( M r 30,400) appeared to be selectively suppressed by bromobenzene treatment. This molecular mass species was the main component of two isoelectric focusing bands with p I 6.23 and 5.91. The effect in vivo of bromobenzene could not be duplicated by in vitro experiments involving treatment of purified enzyme with bromobenzene in the presence of a NADPH-dependent microsomal drug metabolizing system. The phenomenon of multiplicity of the reductase was not limited to the liver. Multiplicity of the enzyme was detected also in the spleen; however, the pattern of composition of the reductase variants vastly differed from that of the liver. In the spleen, variants with p I 5.76, 5.61, and 5.48 were the prevalent forms; the variant with p I 6.23 was absent, and p I 5.91 was present in a minute amount. Further, bromobenzene did not affect the composition pattern of net-charge variants in this organ. Also, the splenic biliverdin reductase activity was refractory to in vivo bromobenzene treatment, whereas the liver reductase activity with both NADH and NADPH was altered by the treatment. The possible significance of the presence of multiple variants of biliverdin reductase and the change in their composition caused by bromobenzene is discussed.

Reduction of delta-aminolevulinate dehydratase concentration by bromobenzene in rats.

The effects of bromobenzene on delta-aminolevulinate (ALA) dehydratase (EC 4.2.1.24) were examined in rats. The enzyme in the bone marrow was irreversibly inhibited after 12 hours of treatment with bromobenzene (intraperitoneally) without any change in the enzyme concentration. When bromobenzene treatment was prolonged to 72 hours, the concentrations of the enzyme in the bone marrow and in the liver were reduced proportionally to the decrease in the enzyme activity. Neither the activity nor the concentration of ALA dehydratase in the peripheral erythrocytes were reduced even after 72 hours treatment with bromobenzene. These findings indicate that bromobenzene decreases ALA dehydratase activity in a biphasic manner; firstly, through an irreversible inhibition probably due to the formation of mercaptide with the essential SH groups and, secondly, through a reduced synthesis of the enzyme.

Cadmium-mediated inhibition of testicular heme oxygenase activity: The role of NADPH-cytochrome c ( P-450) reductase

The concerted activity of two microsomal enzymes, heme oxygenase and NADPH-cytochrome c ( P-450) reductase, is required for isomer-specific oxidation of heme molecule; heme oxygenase is commonly believed to be rate limiting in this activity. In this report, we provide evidence strongly suggesting the rate-limiting role of the reductase in oxidation of heme molecule in rat testis. In the testis and the liver of rats treated with Cd (20 μmol/kg, sc, 24 h) heme oxygenase activity, assessed by the formation of bilirubin, was decreased by 50% and increased by 7-fold, respectively. In these animals, the reductase activity was decreased by nearly 75% in the testis, but remained unchanged in the liver. Similarly, the reductase activity in the liver was not altered when heme oxygenase activity was increased by 20-fold in response to bromobenzene treatment. Addition of purified testicular reductase preparation (purified over 4000-fold), or hepatic reductase, to the testicular microsomes of Cd-treated rats obliterated the Cd-mediated inhibition of heme oxygenase activity. The chromatographic separation of heme oxygenase and the reductase of the testicular microsomal fractions revealed that the reductase activity was markedly decreased (75%) while the heme oxygenase activity, when assessed in the presence of exogenous reductase, was not affected by in vivo Cd treatment. In vitro, the membrane-bound reductase preparation obtained from the testis was more sensitive to the inhibitory effect of Cd than the liver preparation. However, the purified reductase preparations from the testis and the liver exhibited a similar degree of sensitivity to Cd. Based on the molar ratio of heme oxygenase to the reductase in the microsomal membranes of the liver and the testis it appeared that the testicular heme oxygenase, which is predominantly HO-2 isoform, interacts with the reductase less effectively than HO-1; in the induced liver, heme oxygenase is predominantly the HO-1 isoform. It is suggested that due to the low abundance of NADPH-cytochrome c ( P-450) reductase and the apparently lower affinity of the enzyme for HO-2, the reductase exerts a regulatory action on heme oxygenase activity in the testis.

Alteration of hepatic glutathione S-transferases and release into serum after treatment with bromobenzene, carbon tetrachloride, or N-nitrosodimethylamine

The effects of bromobenzene, carbon tetrachloride, and N-nitrosodimethylamine (DMN) on hepatic glutathione S-transferase activity were studied in untreated and in phenobarbital- or ethanoltreated. rats. In phenobarbital-treated rats, the isozymic composition of the hepatic cytosolic glutathione S-transferases was changed after giving hepatotoxic chemicals; glutathione S-transferases 2-2(AA), 3-3(A), 1-2(B), 3-4(C), and 4-4 + 5-5(D+E) were present in cytosol from control rats, but only glutathione S-transferases cochromatographing with transferases 4-4 + 5-5(D+E) were detected in rats given carbon tetrachloride or bromobenzene. A marked decrease in hepatic and an increase in serum glutathione S-transferase activity were also observed after carbon tetrachloride or bromobenzene treatment, but little change was seen after giving DMN. On the contrary, in untreated or ethanol-treated rats. DMN administration decreased hepatic glutathione S-transferase activity and caused an elevation in serum glutathione S-transferase activity. The isozymic composition of the hepatic cytosolic glutathione S-transferases after giving DMN to untreated rats was also altered, but the alteration was much less than that observed after giving carbon tetrachloride or bromobenzene to phenobarbital-treated rats. The elevation in serum glutathione S-transferase activity was accompanied by an increase in both serum glutamate-pyruvate transaminase activity and serum bilirubin concentrations. Thus, hepatic glutathione S-transferase activity was altered and released into serum after giving hepatotoxic chemicals, and the alteration in glutathione S-transferase activity was dependent on treatment with phenobarbital or ethanol.

Subcellular distribution of malathion, phenthoate, and diethylsuccinate carboxylesterases in rat lungs

The subcellular distribution of malathion, phenthoate, and diethylsuccinate carboxylesterases was determined in the lungs of male Sprague-Dawley rats and in rats administered lung toxic doses of bromobenzene and paraquat. In control with the former two substrates, the highest activity was encountered in the 6500 g and 12,000 g pellets. In addition, significant activity was found in the 100,000 g supernatant. These fractions hydrolyzed diethylsuccinate very slowly, and the major diethylsuccinate carboxylesterase activity was recovered in the 100,000 g pellet. The bromobenzene treatment had no effect on these carboxylesterases; however, the paraquat treatment significantly decreased the phenthoate and diethylsuccinate carboxylesterases in the 100,000 g pellet without altering the activity in the other fractions. The present study suggested that the subcellular distribution of malathion and phenthoate carboxylesterases is different from that in liver. The present study revealed that, in the lungs, the highest total activity for malathion and phenthoate carboxylesterases was found in the 100,000 g supernatant, in comparison with liver, where the 100,000 g pellet contains a much higher activity of these enzymes. The decrease in specific activities of diethylsuccinate and phenthoate carboxylesterases following the treatment with a pneumotoxicant may serve as an indicator of lung damage.

Interactions between bromobenzene dose, glutathione concentrations, and organ toxicities in single- and multiple-treatment studies

A single oral dose of 4.0 mmol/kg bromobenzene transiently depleted hepatic and renal reduced nonprotein sulfhydryl group (NPS) concentrations, caused hepatocellular necrosis, and increased serum glutamic-pyruvic transaminase activity in male Fischer 344 rats. The depletion of NPS had partially reversed by 24 hr, and NPS concentrations were approximately twice normal values by 48 hr post-treatment. When the effects of single and repeated (once daily for 2, 4, or 10 days) treatments with 4.0 mmol/kg were compared, it was apparent that the severity of hepatotoxicity lessened and the percentage depletions of hepatic and renal NPS concentrations decreased with increasing length of bromobenzene treatment. There were essentially no signs of toxicity following the tenth treatment with 4.0 mmol/kg. Single-treatment studies indicated the following dose-response: 2.0 mmol/kg bromobenzene depleted liver NPS and was hepatotoxic, 0.5 mmol/kg caused a lesser depletion of liver NPS and was not (overtly) hepatotoxic, and 0.0625 mmol/kg was the maximum dose that did not deplete liver NPS. The responses to single and multiple (ten) treatments with these representative doses were compared. Liver injury was observed after a single but not after the tenth daily treatment with 2.0 mmol/kg. Both the single and the tenth administrations of 2.0 mmol/kg depleted hepatic NPS, but the precentage of depletion was greater after the first than after the tenth dose. Liver injury was not detected with lower dose regimens. The patterns of NPS depletion in liver and kidney were similar after single or muliple (ten) treatments. The minimum NPS concentrations produced, however, were lower after single than after multiple treatments. The molar amounts of liver NPS depleted after the tenth treatment appeared to be equivalent to or greater than those after the first, but prior bromobenzene exposure resulted in a higher concentration of tissue NPS being present at the time of the final treatment. Thus, the minimum tissue concentrations of NPS were greater after multiple treatments than after single treatments, despite the loss of equivalent amounts of NPS. It is concluded from these studies that repeated treatment produces resistance to bromobenzene hepatotoxicity. This protective adaptation may be due to a chemically induced increase in liver glutathione concentration.