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Induction of micronuclei in rat bone marrow after chronic exposure to lead acetate trihydrate
Abstract
Lead increasingly contributes to pollution of the environment and may play a role in the development of adverse effects in the human and animal body. Data concerning its mutagenic, clastogenic, and carcinogenic properties have been conflicting. In this study, we evaluated the frequency of micronuclei in bone marrow erythrocytes of rats treated with lead acetate trihydrate. Outbred Wistar rats were exposed to a daily dose of 100 mg/L drinking water for 125 days. The mean value of the total number of micronuclei observed in polychromatic erythrocytes of female rats was significantly higher than that found in the control group (13.375 +/- 2.722 against 9.625 +/- 3.204 micronuclei/1000 cells; P = 0.024 in ANOVA). In exposed female animals, no significant reduction of the ratio of polychromatic to normochromatic erythrocytes was observed (0.990 +/- 0.228 against 1.208 +/- 0.195; P = 0.060 in ANOVA). The effects of lead acetate trihydrate in male rats are both cytotoxic and genotoxic because of a decrease in ratio of polychromatic to normochromatic erythrocytes (0.715 +/- 0.431 against 1.343 +/- 0.306; P = 0.023, ANOVA followed by Tukey test) and an increase in frequency of micronucleated polychromatic erythrocytes (24.167 +/- 7.859 against 4.0 +/- 4.528 micronuclei/1000 cells; P < or = 0.001, ANOVA followed by Tukey test), respectively.
... Manganese is an essential element for the living being and at the same time a toxic metal, which is considered one of the most abundant elements on earth's crust, therefore we can find it in rocks, soil, water, and food (Lebdaet al., 2012).Nonetheless, this metal is toxic at many levels, mainly neurotoxic which induce symptoms similar to those of Parkinson with serious repercussions on numerous organs like the liver, kidney (Racette et al., 2012). Lead and manganese can also alter the anti-oxidative status of the cells (Alghazal, et al., 2008). ...
The mechanisms in the origin of lead and manganese poisoning (Pb-Mn) are multiple, and potentially touch all the body cells. For this purpose, we are interested in oxidative stress induced by Pb-Mn at the erythrocytic level at a dose of 0.2% (Pb) and 4.79 mg/ml (Mn) during the period of gestation and lactation in wistars rats and the ability of mint essential oil, Mentha spicata (MEO) to restore or not this state of stress. The extraction of MEO "HEM" via steam distillation allows us to obtain MEO with an output of 0.49%. Moreover , the analysis of the anti-oxidativeerythrocytic status showed that the co-exposure to Pb-Mn significantly increases enzymes activity of catalase (CAT) and superoxide dismutase (SOD), and significantly reduces the enzyme activity of glutathione peroxidase (GPx), by causing a dysfunction in the anti-oxidative defense system. In contrast, the administration of MEO via intra-peritoneal (IP) during a period of 21 days in the previously Pb-Mn intoxicated rats, that MEO contributes significantly in the improvement of the defenses against radical aggression, through a recovery at the level of anti-oxidative enzymes activities by increasing their ability to eliminate radical compounds.
... However, the possible relationship of genotoxicity and specific metal contamination is still questionable. Although this association is indicated in studies with different organisms and metal concentrations, whether it is lead (Çelik et al., 2005;Piao et al., 2007;Alghazal et al., 2008;Tapisso et al., 2009;García-Lestón et al., 2010), copper (Franke et al., 2006;Serment-Guerrero et al., 2011), chrome (Papageorgiou et al., 2008), cadmium (Seoane and Dulout, 2001;Bertin and Averbeck, 2006;Ahmed et al., 2010;Otomo and Reinecke, 2010), manganese (Gauthier et al., 2004;Erbe et al., 2011) andmercury (Al-Sabti andMetcalfe, 1995;Porto et al., 2005;Cavas, 2008). According to Luoma and Rainbow (2008), the hierarchy of toxicity by metals analyzed in the present study on aquatic organisms is as follows: Hg > Cu > Cd > Pb > Cr. ...
Globally, there is a lack of knowledge about tropical ecotoxicology dealing with the potential impact of metal contamination in mangrove ecosystem. This habitat is considered a nursery for several animal species, among them the “uçá”-crab (Ucides cordatus), known as a key species due to its biological and economical importance. This study evaluated the association involving metal contamination (Cd, Cu, Pb, Cr, Mn and Hg) in water, sediment, red-mangrove vegetation (Rhizophora mangle) and tissues of uçá crab, together with its geno-cytotoxic responses, based on micronucleated hemocytes frequency and the retention time of neutral red in lysosomes. We assessed six mangrove areas with distinct pollution levels in São Paulo State, Brazil, where the water and sediment contamination by metals were associated with accumulation of these pollutants in biotic compartments (mangrove leaves and crab). In U. cordatus, metal accumulation was best explained by metal concentration found in leaves of R. mangle than in the water or sediment, indicating that feeding drives metal exposure in this organism. Mercury (Hg) concentration in sediment, copper (Cu) concentration in hepatopancreas of U. cordatus and lead (Pb) in water and green leaves of R. mangle showed a significant correlation with genotoxic impact in U. cordatus. However, copper concentration (in green/senescent leaves and hepatopancreas) and lead (in sediment), were the major metals affecting lysosomal membrane integrity. Therefore, representatives of all compartments were associated with cyto and genotoxicity in this species, thus requiring a holistic approach to issues related to sublethal damage. Probability estimates of cytogenetic impacts related to metal concentration in abiotic compartments (significantly correlated with known biomarkers: Hg in sediment; and Pb in water and sediment) are also presented. Our results highlight the need for environmental restoration of mangroves areas contaminated with metals, responsible for cytogenetic injuries and revealing a pre-pathological condition in this sentinel species, in addition to ecological disturbances.
... Leukocytosis, monocytosis and neutrophilia were observed at higher doses of administered lead (Okediran et al., 2010). Similar observation was reported by some scholars with respect to the toxicological effect of lead on blood (Drugut et al., 2008;Ibrahim et al., 2011;Alghazal et al., 2008;Nashwa et al., 2006;Ayla et al., 2005;Banu et al., 2007). ...
Lead is a heavy metal which is reported to have toxicological effects on various organs in humans and animals. Due to its toxicological effects this study reviews current literature on the toxicological profile of lead on the testis, bone, blood and lungs and the possible roles of mitigating agents. In this study reports showed that in the brain lead induced cerebellar edema, cerebral satellitosis and encephalomalacia with impairments in cortex, hippocampus and cerebellum. Lead impairment of haematological parameters manifested as decrease in packed cell volume, haematoglobin, red blood cell count and total erythrocyte count. Anemia, leucopenia, neutropenia, lymphopenia and monocytopenia were also reported. Lead treated testis was characterized by reduction in the weight of sex glands, testicular sperm counts, daily sperm production, sperm density, sperm viability, sex hormones with increase in sperm abnormalities. Histopathological study of lead treated testis revealed loss of germ cells with pyknotic nuclei and vacuolated cytoplasm. Apoptosis of sertoli cells, leydig cells and mitochondria degeneration were reported. Lead treated lungs were characterized by mononuclear cell proliferation, mononuclear cell invasion, collagen fibre accumulation in the interalveolar septa and pneumonia. The antioxidants status of these organs were impaired making these organs vulnerable to lead induced oxidative stress. This laid credence to the generation of reactive oxygen species as one of the mechanisms of lead induced toxicological effects in various organs. In this study it was also observed that the toxicological effects of lead were mitigated by vitamin C, vitamin E, selenium, zinc and calcium. Chemical agents like melanin, casein, DMSA and CaNa2-EDTA also mitigated lead induced toxicity. Some extracts of plant origin also ameliorated the toxicological effects of lead. Some of these mitigating agents may require further evaluation if they could be of clinical application.
... Additionally, Aboul-Ela [9] detected a significant increase of structural chromosomal aberrations in bone marrow cells and primary spermatocytes of albino mice treated with LA. In addition, LA proved to be a potent micronuclei inducer in vivo and in vitro test systems: LA induced micronuclei in kidney cells of Sprague-Dawley albino rats [10]; in human melanoma cell [11]; in Chinese hamster V79 cells [12,13]; in Wistar rats' leukocytes, reticulocytes and erythrocytes [14,15]; in rats' erythrocytes [16][17][18]; in peripheral blood erythrocytes tissues, gill and fin epithelial cells of Carassius auratus [19] and in bone marrow cells of Algerian mice [20]. More so, the results of the studies indicated that occupational exposure to Pb has adverse effects on sperm parameters (decreased sperm counts, lower and a lesser motility and altered sperm morphology), studies showed that exposure to inorganic lead greatly impaired male reproductive functional activities by decreasing sperm count or distorting sperm motility and morphology [21]. ...
... Additionally, Aboul-Ela (2002) detected a significant increase of structural chromosomal aberrations in bone marrow cells and primary spermatocytes of albino mice treated with LA. In addition, LA proved to be a potent micronuclei inducer in vivo and in vitro test systems: LA induced micronuclei in kidney cells of Sprague–Dawley albino rats (Robbiano et al., 1999); in human melanoma cell (Poma et al., 2003); in Chinese hamster V79 cells (Thier et al., 2003; Bonacker et al., 2005); in Wistar rats' leukocytes, reticulocytes and erythrocytes (Kasˇuba et al., 2004; C ¸ elik et al., 2005); in rats' erythrocytes (Piao et al., 2007; Alghazal et al., 2008); in peripheral blood erythrocytes, gill and fin epithelial cells of Carassius auratus auratus (C ¸ avas, 2008) and in bone marrow cells of Algerian mice (Tapisso et al., 2009). Moreover, positive results indicate the induction of SCE with LA was obtained by Poma et al. (2003) in human melanoma cells and in bone marrow cells of Algerian mice (Tapisso et al., 2009). ...
Lead represents a significant ecological and public health concern due to its toxicity and its ability to accumulate in living organisms. The present investigation was designated to assess the modulating effect of melatonin (MLT) against lead acetate (LA) genotoxicity. Three cytogenetic end points were considered: the frequencies of micronucleated polychromatic erythrocyte cells (MnPCEs) in the bone marrow, the chromosomal aberration in the primary spermatocytes and the frequency of sperm abnormalities. Male mice were used in this experiment; animals were divided into 8 groups of 6 animals each. First group received an oral gavage of solvent (4% ethanol) and served as control and the other groups received an oral gavage of MLT (10 mg/kg) and/or 10, 50, 100 mg/kg body weight of lead acetate for 11 days. Mice were scarified 24 h after the last treatment. Examination and analysis of MnPCEs and meiotic metaphases showed no mutagenic effect of melatonin. Meanwhile LA induced a significant (P < 0.01) increase in the three end points used in this investigation. Results showed that melatonin caused a significant reduction in MnPCEs and chromosomal aberrations in meiotic cells. PCE/NCE ratios in bone marrow also increased in relation to melatonin and LA treatments. Moreover, melatonin decreased the % of sperm abnormality by 28.3%. The data obtained in this study suggest that melatonin administration confers protection against damage inflicted by LA, and support the contention that melatonin protection is achieved by its ability as a scavenger for free radicals generated by LA.
... Nudi et al. (2010) mentioned that the uçá-crab is not efficient as a PAH biomarker under field conditions, although it is known that organic pollutants can cause genetic impacts in many species. Although recent studies revealed significant genetic changes caused by metals during exposure at different concentrations, including lead (Celik et al. 2005; Piao et al. 2007; Alghazal et al. 2008; Tapisso et al. 2009; García-Lestón et al. 2010), copper (Franke et al. 2006; Serment-Guerrero et al. 2011), chromium (Papageorgiou et al. 2007;, and cadmium (Seoane and Dulout 2001; Bertin and Averbeck 2006; Ahmed et al. 2010; Otomo and Reinecke 2010). In the Cubatão mangroves, metal contamination was detected in concentrations below TEL and PEL levels. ...
In Brazil, the state of São Paulo contains both preserved areas (Juréia-Itatins Ecological Station) and extremely impacted ones (Cubatão Municipality). This study evaluated the concentrations of five metals (Cu, Cd, Cr, Pb, and Hg) in two mangroves with different levels of anthropogenic impact and the apparent genotoxicity to Ucides cordatus. Water and sediment samples were obtained, and metal concentrations were determined with an atomic absorption spectrophotometer. The genotoxic impact was quantified based on the number of micronucleated cells per 1,000 analyzed (MN‰), using hemolymph slides stained with Giemsa. Metal concentrations in water were below the detection limit, except for lead, although no significant difference was observed between the areas (P > 0.05). Sediment from Cubatão had higher concentrations of Cd, Pb, Cr, and Cu than sediment from Juréia-Itatins (P < 0.05), but no significant differences in metal concentrations were detected among depth strata of the sediment (P > 0.05). Crabs from Cubatão had a 2.6 times higher mean frequency of micronucleated cells (5.2 ± 1.8 MN‰) than those from Juréia-Itatins (2.0 ± 1.0 MN‰; P < 0.0001). The more-polluted conditions found in the mangrove sediments of Cubatão were reflected in the micronucleus assay, demonstrating their genotoxic effect; however, genetic damage should be attributed to a synergistic effect with other kinds of pollutants previously recorded in different environments of Cubatão. U. cordatus proved to be an excellent bioindicator of mangrove pollution. This study established, for the first time, the normal frequency of MN‰ in a population of this species within an ecological station.
... Heavy metals can cause a wide scale of toxicological and biochemical effects (Lukáč et al., 2009), and they can cause damage to a lot of animal and human organs and tracts (Alghazal et al., 2008;Gurer-Orhan et al., 2004). The current experiments consider that the formation of reactive forms of oxygen is the main pathogenetic mechanism of metal-induced carcinogenesis (Valko et al., 2006) with extra accent to their role in signal traces activation, epigenetic changes and DNA reparation processes (Salnikov and Zhitkovich, 2008). ...
The aim of the study was to assess the effects of exposure to low doses of lead dissolved in drinking water (average daily dose of 2.2 mg kg(-1) day(-1)) on selected carbohydrate metabolism parameters in 20 wistar rats. Animals were divided into two groups - control (C) (group drinking clear water) and experimental group (Pb; group exposed to low doses of lead acetate in a concentration of 100 μmol l(-1) of drinking water). In this study, we studied the biochemical parameters (glucose, haemoglobin (Hb), glycated haemoglobin (HbA1c), lactate dehydrogenase (LDH) and amylase (AMS)) in rat blood. Glucose and Hb concentration and AMS activity decreased, LDH activity increased but HbA1c concentration levels did not change in rats exposed to lead. Our results well documented that lifetime exposure to lead affected carbohydrate metabolism of rats. Some parameters like concentration of Hb as well as activities of AMS and LDH are useful markers of intoxication of rats with lead. For the evaluation of results (e.g. AMS), not only the data at the end of the experiment should be taken into account but also the entire duration of trials (i.e. more time steps) that makes results more objective should be considered.
... The frequency of micronuclei can be most easily evaluated in young erythrocytes shortly after the main nucleus is expelled. These young erythrocytes are termed as polychromatic erythrocytes (PCE) and are distinguished from the mature normochromatic (NCE) erythrocytes.[56] Chromosomal aberrations are carried out to evaluate the frequency of structural chromosomal aberrations like gaps, breaks and translocations. ...
The ORMO-48 is a new indigenous material for dental applications, developed by the Dental Products Laboratory of our Institute. The aim of the present study was to evaluate the genotoxic effect of an indigenously developed dental material in Swiss albino mice. The genotoxic effect was evaluated by micronucleus and chromosomal aberration tests. Two grams of dental material was extracted in 10.0 ml of physiological saline at 70°C for 24 h. The extract was cooled to room temperature and was used for the experiment. The experimental designed had three groups each (six mice in each group) for micronucleus and chromosomal aberration tests. The first, second, and third groups were given a single exposure of physiological saline alone (control), dental material's extract (test), and cyclophosphamide (positive control) respectively for micronucleus and chromosomal aberration tests. The result of the study indicated that, the percentage of micronucleated PCE (polychromatic erythrocytes) and NCE (normochromatic erythrocytes) induced by the dental material (extract) treated group was well comparable with control group, whereas the positive control induced significantly high (P < 0.001) micronucleated PCE when compared to control. The PCE and NCE ratio of the dental material extract treated group was similar to that of control group. The chromosomal anomalies such as chromatid/chromosomal breaks, centric rings, exchanges, dicentric, and acentric fragments were evaluated. The result showed that the anomalies of the dental material extract treated group were similar to control group, however, significant anomalies were observed in the cyclophosphamide treated group. Hence, the present study concluded that the indigenously developed biocompatible dental material, ORMO-48 is non genotoxic at our laboratory conditions.
... Several studies that evaluated the genotoxic effects of lead acetate in rodents by means of the MN test showed an increase in the frequency of MN (Tachi et al., 1985;Robbiano et al., 1999;Çelik et al., 2005;Piao et al., 2007;Tapisso et al., 2009). Alghazal et al. (2008) analysed the MN rate in bone marrow erythrocytes of male and female Wistar rats treated with lead acetate trihydrate. They found a significant increase in the total number of MN in polychromatic erythrocytes of both male and female rats with regard to the control group. ...
Lead is a ubiquitous toxic heavy metal with unique physical and chemical properties that make it suitable for a great variety of applications. Because of its high persistence in the environment and its use since ancient times for many industrial activities, lead is a common environmental and occupational contaminant widely distributed around the world. Even though the toxic effects of lead and its compounds have been investigated for many years in a variety of systems, the data existing with regard to its mutagenic, clastogenic and carcinogenic properties are still contradictory. The International Agency for Research on Cancer has classified lead as possible human carcinogen (group 2B) and its inorganic compounds as probable human carcinogens (group 2A). Furthermore, although the biochemical and molecular mechanisms of action of lead remain still unclear, there are some studies that point out indirect mechanisms of genotoxicity such as inhibition of DNA repair or production of free radicals. This article reviews the works listed in the literature that use different parameters to evaluate the genotoxic effects of lead in vitro, in vivo and in epidemiological studies.
Lead, a highly toxic pollutant, causes numerous health problems and affects nearly all biological systems thus arousing interest in using antioxidants to reduce its toxic effects. Therefore, the undertaken study estimated the influence of cerium oxide nanoparticles (CeO2-NPs) on the lead acetate–induced genotoxicity and inflammation in the kidney and heart tissues of mice. Twenty male mice were randomly divided into negative control and lead acetate and/or CeO2-NPs administrated groups. Comet and diphenylamine assays were conducted to assess the DNA damage and the expression of apoptosis-related genes and inflammatory cytokines were also measured in addition to the estimation of reactive oxygen species (ROS) level. Co-administration of CeO2-NPs significantly reduced the DNA damage and ROS generation caused by lead acetate in the kidney and heart tissues. The co-administration of CeO2-NPs also ameliorated the lead acetate–induced dysregulation in the expression levels of p53, K-ras, interleukin-6, and cyclooxygenase-2 in the kidney and heart. Conclusion: the co-administration of CeO2-NPs suppresses the genotoxicity, inflammation, and ROS generation resulting from lead acetate administration and restoring the genomic DNA integrity; thus, administration of CeO2-NPs is recommended to minimize the lead acetate–induced hazards.
Our work focused on the impact of chronic lead acetate and manganese chloride poisoning on renal function in Wistar rats. It also tested the efficacy of the essential oil extracted from (Mentha spicata) spearmint in reversing these effects by treating previously poisoned rats with a daily intraperitoneal injection of 0.1 ml HEM/kg over a 21-days period. Analysis of this essential oil through gas chromatography coupled with mass spectrometry revealed that the main components are: carvone (42.2%), menthone (20.89%), piperitenone (17.41%) and isomenthone (7.99%). Chronic oral co-exposure to lead and manganese during periods of gestation and lactation resulted in a significant reduction (P<0.01) in both body and kidney weight. Further examination of renal function revealed significant disturbances in the quantity of markers of renal function (creatinine, urea, uric acid) in the blood. Chronic co-exposure led to the observation of disruption to antioxidant enzyme (superoxide dismutase, glutathione peroxidase and catalase) activity in poisoned rats in comparison with the control rats; these results suggest renal failure. Indeed, histological study of the kidneys revealed pronounced damage manifesting as cell degeneration with the renal parenchyma affected by inflammation. In addition, the administration of the essential oil of Mentha spicata led to the observation of an increase in body weight, regulation of the various biochemical markers, and the regeneration of damaged renal tissue, all of which proves the high importance of spearmint in traditional medicine, and its therapeutic properties in combating renal problems.
The antioxidant and antiproliferative properties of mango by-products were investigated. This study was carried out to evaluate the protective role of mango peel or kernel defatted extracts against Pb-acetate adverse effects on oxidant/antioxidant status, liver dysfunction biomarkers, histopathological changes and genotoxicity in male mice. Total phenolic content and antioxidant activity of both extracts were evaluated. Two doses of both extracts (50 and 100 mg/kg) were used to evaluate their role against the toxicity of Pb-acetate (500 ppm). Mice given mango extracts with Pb-acetate had significantly lower plasma MDA, AST and ALT and higher glutathione than mice given Pb-acetate alone. Mango extracts prevented the histopathological changes in liver induced by Pb-acetate and decreased the cytotoxicity of lead by increasing the ratio of PCE/NCE. Mango extract treatment reduced the DNA damage induced by Pb-acetate in liver as demonstrated by a reduction in micronuclei and decrease in tail length, tail DNA% and Olive tail moment. It can be concluded that mango by-product extracts have potential to protect from oxidative stress and genotoxicity of lead.
Lead is a common industrial and environmental pollutant. Prolonged exposure of a sublethal dose to this toxicant is associated with oxidative stress, damage of DNA and considered to be a risk factor for kidney, liver added to many disorders. This study was carried out to investigate the most toxic effects of lead with trial to diminish this toxicity by supplementation of casein or activated charcoal. Therefore, forty mature male albino rats were used, they divided into four equal groups, 10 rats of each. Group 1 was considered as control. Rats of other groups (2, 3 and 4) were supplemented lead acetate at a dose of 0.5 g/100 ml drinking water for 2 months. Gr 3 is given charcoal at a concentration of 0.05g ration, while in Gr 4 casein is mixed with ration at a concentration of 20 g/100 g ration. Group 2 revealed significant increases in serum AST, ALT, ALP, LDH, GGT, TB, TC, LDL and CAT levels and decreased total protein, HLDL, VLDL, trace mineral salts, SOD and GSH- px. The results of haematological study evoked a significant decrease in the red blood cells (RBCs) count, blood haemoglobin (Hb) concentration and packed cell volume (PCV). Cytogenetic study resulted a significant increase in the percent of multinucleated polychromatic erythrocytes (MPCE) as well as significant increase in the polychromatic- erythrocytes (PCE) and normochromatic erythrocyte (NCE) ratio (PCER/NCE) Lead concentration in level serum and different tissues showed lead accumulation in serum, kidney, liver, muscle, intestine and spleen. Supplementation of charcoal and casein induce protective effects against lead toxicity in most of above parameter studies, and casein showed higher efficacy in the protection of lead- induced toxicity than charcoal.
Objective: Several pollutants represent a significant ecological and public health concern due to their toxicity and their ability to accumulate in living organisms in which lead is one of them. The present investigation was designated to assess the modulating effect of thymoquinone (TQ) against lead acetate (LA) toxicity. Methods: Several endpoints were considered to design this study such as: The gene expression of tumor initiation genes (cytochrome P450 3A [CYP3A], cyclooxygenase 2 [COX2], BAX and Bcl2), DNA damage and alterations in the levels of glutathione (GSH), lipid oxidation (malondialdehyde [MDA]), and protein oxidation (protein carbonyl [PC]) in male rats. About 60 male rats were used in this study which allocated in six groups (10 animal each) and treated with LA (200 mg/kg diet), TQ (5 and 10 mg/kg b.wt.), and LA + TQ. Results: The results revealed that LA induced significant DNA damage and alteration in the expression of CYP3A, COX2, BAX, and Bcl2 as well as induced changes in GSH content and MDA and PC levels in male rats. Meanwhile, TQ was decreased significantly the toxic effect of LA in male rats which decreased the alterations in the gene expression and DNA damage as well as GSH content and MDA and PC levels. Conclusion: The results suggested that TQ treatment confers protection against toxicity inflicted by LA and support the contention that TQ protection is achieved by its ability as a scavenger for free radicals generated by LA. © 2015, Asian Journal of Pharmaceutical and Clinical Research. All rights reserved.
The antioxidant and antiproliferative properties of mango by-products were investigated. This study was carried out to evaluate the protective role of mango peel or kernel defatted extracts against Pb-acetate adverse effects on oxidant/antioxidant status, liver dysfunction biomarkers, histopathological changes and genotoxicity in male mice. Total phenolic content and antioxidant activity of both extracts were evaluated. Two doses of both extracts (50 & 100 mg/kg) were used to evaluate their role against the toxicity of Pb-acetate (500ppm). Mice given mango extracts with Pb-acetate had significantly lower plasma MDA, AST and ALT and higher glutathione than mice given Pb-acetate alone. Mango extracts prevented the histopathological changes in liver induced by Pb-acetate and decreased the cytotoxicity of lead by increasing the ratio of PCE/NCE. Mango extract treatment reduced the DNA damage induced by Pb-acetate in liver as demonstrated by a reduction in micronuclei and decrease in tail length, tail DNA% and olive tail moment. It can be concluded that mango by-product extracts have potential to protect from oxidative stress and genotoxicity of lead.
In the present study, the single and combined neurotoxic effects of benzo[a]pyrene ([BaP] 0.0, 0.5, and 5.0 mg/kg body weight; intragastric administration) and lead acetate (0.0, 5.4, and 54.0 mg/L; by drinking water) were examined on KunMing mice. In the Morris water maze, results showed that BaP and lead induced synergistic effects on the escape latency and the time spent in the target quadrant but also showed an additive effect on the number of times animal crossing the original platform. Also, BaP and lead induced a synergistic effect on DNA damage in the single-cell gel electrophoresis. However, BaP plus lead showed additive effects on the levels of malondialdehyde, superoxide dismutase, and glutathione. These results suggested that the combination of BaP and lead can lead to a synergistic effect on spatial learning and memory impairments, and the mechanisms of the synergistic effects on behavioral deficits may be due to the oxidative stress injury.
In recent years, adverse health effects of chemicals from electronic waste (e-waste) have been reported. However, little is known about the genotoxic effects of chemicals in e-waste. In the present study, air concentrations of the toxic metals at e-waste and control sites were analyzed using inductively-coupled plasma mass spectrometry. Levels of toxic metals (lead, copper and cadmium) in blood and urine were detected using atomic absorption spectrophotometry in 48 exposed individuals and 56 age- and sex-matched controls. The frequencies of lymphocytic micronucleated binucleated cells (MNBNCs) were determined using a cytokinesis-block micronucleus assay. Results indicated that blood lead levels were significantly higher in the exposed group (median: 11.449 μg/dL, 1st/3rd quartiles: 9.351-14.410 μg/dL) than in the control group (median: 9.104 μg/dL, 1st/3rd quartiles: 7.275-11.389 μg/dL). The exposed group had higher MNBNCs frequencies (median: 4.0 per thousand, 1st/3rd quartiles: 2.0-7.0 per thousand) compared with the controls (median: 1.0 per thousand, 1st/3rd quartiles: 0.0-2.0 per thousand). Additionally, MNBNCs frequencies and blood lead levels were positively correlated (r = 0.254, p<0.01). Further analysis suggested that a history of working with e-waste was a predictor for increased blood lead levels and MNBNCs frequencies in the subjects. The results suggest that both the living and occupational environments at the e-waste site may be risk factors for increased MNBNCs frequencies among those who are exposed.
The mutagenic and carcinogenic potentials of lead are still being investigated. The aim of this study was to evaluate the genotoxic effect of lead acetate in the early period of life, when the organism is extremely sensitive to toxic effects of lead. Six-day-old suckling Wistar rats were exposed to lead (as acetate) either orally for 9 days (daily dose 2 mg Pb/kg b. wt., 18 mg/kg b.wt. total dose) or by a single intraperitoneal injection (5 mg Pb/kg b. wt.). DNA damage was investigated using the comet assay and in vivo micronucleus test. The results of the comet assay showed statistically significant differences between the control (unexposed) animals and the two groups of exposed animals by ANOVA weighted for unequal variance (heterogeneity of variances was found by Levene's test), followed by Tukey's post hoc test at the level of significance of p < 0.05. The two groups of lead-exposed animals were also significantly different from each other. Orally lead-exposed animals showed a significant increase of micronuclei frequencies in reticulocytes and erythrocytes compared to unexposed animals (ANOVA, p < 0.05).
High-dose lead exposure in rodents has been shown to produce pathognomonic lead intranuclear inclusion bodies and to result in an increased incidence of renal adenocarcinomas. Studies from this laboratory and others have demonstrated the presence of high-affinity renal lead-binding proteins in rat kidneys which act as tissue sinks for lead at low dose levels. Cell-free nuclear translocation studies have shown that these molecules are capable of facilitating the intranuclear movement of lead and that they are associated with chromatin. These data suggest that renal lead-binding proteins may play a role in mediating known alterations in renal gene expression associated with formation of intranuclear inclusion bodies. More recent studies from this laboratory have demonstrated the presence of chemically similar lead-binding proteins in kidneys of both monkeys and humans. Such observations suggest that a similar mechanism may be operating in primates since lead intranuclear inclusion bodies are also observed in these species. These data provide a testable mechanistic approach for assessing the possible role(s) of lead-binding proteins in mediating the intranuclear movement of lead and lead-induced renal cancer in primate species.
Even though the toxic effects of lead and cadmium compounds have been studied over many years, inconsistent results have been obtained about their mutagenic, clastogenic and carcinogenic properties. However, these metals are considered to be potential human carcinogens. The mechanism of metal-induced carcinogenesis is still unknown, but one possible pathway may involve the interaction of metals with DNA, either directly or indirectly. In this work we explore the capacity of lead, cadmium or a mixture of both metals to interact with acellular DNA, by employing a variant of the comet assay. Our results, using low non-cytotoxic metal concentrations (0.01, 0.1 and 1.0 microM) with the standard protocol for the acellular assay, showed an induction of DNA damage in cells of all organs studied; however, basal DNA damage was different in each organ. To confirm that we were working with pure DNA, proteinase K was added to the lysis solution. With this enriched-lysis solution we found a negative response in the induction of DNA damage in cells derived from the liver, kidney and lung of CD-1 male mice. To support the results obtained by the enriched-acellular assay, we studied the capacity of lead and cadmium (0.1 microM) to induce breaks in pooled genomic DNA in cells of the same organs, with negative results. Consistent with these findings, these metals do not induce DNA breaks in the plasmid pUSE amp+. On the whole, we did not detect direct induction of DNA strand breaks by lead acetate, cadmium chloride or a mixture of both metals, all at low non-cytotoxic concentrations. However, we found an induction of lipid peroxidation and an increase in free radical levels in the different organs of CD-1 male mice after inhalation of lead acetate (0.0068 microg/cc) or cadmium chloride (0.08 microg/cc) for 1 h, suggesting the induction of genotoxicity and carcinogenicity by indirect interactions, such as oxidative stress.
Lead is perhaps the longest used and best recognized toxic environmental chemical and it is still being used recklessly. Lead (Pb) has been found to be capable of eliciting a positive response in an extraordinarily wide range of biological and biochemical tests; among them tests for enzyme inhibition, fidelity of DNA synthesis, mutation, chromosomal aberrations, cancer and birth defects. Since inhalation is one of the most important routes of environmental Pb exposure, in the present study a lead inhalation model in mice was implemented in order to detect the induction of genotoxic damage as single-strand breaks and alkali-labile sites in several mouse organs (nasal epithelial cells, lung, whole blood, liver, kidney, bone marrow, brain and testes), assessed by single cell gel electrophoresis (SCGE) or Comet assay. We found differences among the organs studied after a single and subsequent inhalations: in the organs analyzed we observed a positive induction of DNA damage after a single inhalation only in the liver and the lung. In subsequent inhalations the response was positive in all organs except the testicle, however, DNA damage induction over time was different for each organ. A correlation between length of exposure, DNA damage and metal tissue concentration was observed for lung, liver and kidney. Differences in DNA damage occurred in organs when lead acetate was administered acutely or sub-chronically. These results show that lead acetate inhalation induces systemic DNA damage but that some organs are special targets of this metal, such as lung and liver, depending in part on length of exposure, suggesting alternative organ processes to handle lead intoxication.
Co-exposure to cadmium, cobalt, lead and other heavy metals occurs in many occupational settings, such as pigment and batteries production, galvanization and recycling of electric tools. However, little is known about interactions between several heavy metals. In the present study we determined DNA single strand break (DNA-SSB) induction and repair capacity for 8-oxoguanine in mononuclear blood cells of 78 individuals co-exposed to cadmium (range of concentrations in air: 0.05-138.00 micro g/m(3)), cobalt (range: 0-10 micro g/m(3)) and lead (range: 0-125 micro g/m(3)). Exposure to heavy metals was determined in air, blood and urine. Non-parametric correlation analysis showed a correlation between cadmium concentrations in air with DNA-SSB (P = 0.001, R = 0.371). Surprisingly, cobalt air concentrations correlated even better (P < 0.001, R = 0.401), whereas lead did not correlate with DNA-SSB. Logistic regression analysis including 11 possible parameters of influence resulted in a model showing that cobalt in air, cadmium in air, cadmium in blood and lead in blood influence the level of DNA-SSB. The positive result with cobalt was surprising, since exposure levels were much lower compared with the TRK-value of 100 micro g/m(3). To examine, whether the positive result with cobalt is stable, we applied several logistic regression models with two blocks, where all factors except cobalt were considered preferentially. All strategies resulted in the model described above. Logistic regression analysis considering also all possible interactions between the relevant parameters of influence finally resulted in the following model: Odds ratio = 1.286(Co in air) x 1.040(Cd in air) x 3.111(Cd in blood) x 0.861(Pb in air) x 1.023(Co in air x Pb in air). This model correctly predicts an increased level of DNA-SSB in 91% of the subjects in our study. One conclusion from this model is the existence of more than multiplicative effects for co-exposures of cadmium, cobalt and lead. For instance increasing lead air concentrations from 1.6 to 50 micro g/m(3) in the presence of constant exposures to cobalt and cadmium (8 micro g/m(3) and 3.8 micro g/m(3)) leads to an almost 5-fold increase in the odds ratio, although lead alone does not increase DNA-SSB. The mechanism behind these interactions might be repair inhibition of oxidative DNA damage, since a decrease in repair capacity will increase susceptibility to reactive oxygen species generated by cadmium or cobalt. Indeed, repair of 8-oxoguanine decreased with increasing exposures and inversely correlated with the level of DNA-SSB (P = 0.001, R = -0.427). Protein expression patterns of individuals exposed to cobalt concentrations of approximately 10 micro g/m(3) were compared with those of unexposed individuals using two-dimensional gel electrophoresis. Qualitative and apparent quantitative alterations in protein expression were selective and certainly occurred in <0.1% of all proteins. In conclusion, the hazard due to cobalt exposure - that has been classified only as IIB by the IARC - seems to be underestimated, especially when individuals are co-exposed to cadmium or lead. Co-exposure may cause genotoxic effects, even if the concentrations of individual heavy metals do not exceed TRK-values.
The data concerning the mutagenic, clastogenic and carcinogenic properties of inorganic lead compounds have been conflicting. Here, we evaluated the frequency of micronuclei in the peripheral blood of female rats treated with three different lead acetate doses. Outbred female Wistar rats were treated by gavage once per week for 10 weeks with cumulative doses of 140, 250 and 500 mg/kg body weight (body wt) of lead acetate. Mitomycin C (MMC) 2 mg/kg body wt was used as a positive control. The aim of the present study was to investigate the possible cytotoxic and genotoxic effects of lead acetate on peripheral blood reticulocytes using the micronucleus test following chronic exposure. The results show the effects of lead acetate in peripheral blood reticulocytes. These effects are both cytotoxic and genotoxic because of a decrease in the number of polychromatic erythrocytes in the peripheral blood and an increase in frequency of micronucleated reticulocytes, respectively.
Environmental lead exposure is associated with reduced bone growth and quality, which may predispose to osteoporosis. Zinc supplementation may reduce lead accumulation; however, effects on bone development have not been addressed. Our objective was to investigate the effects of marginal zinc (MZ) and supplemental zinc (SZ) intakes on bone lead deposition and skeletal development in lead-exposed rats. In a factorial design, weanling Sprague-Dawley rats were assigned to MZ (8 mg/kg diet); zinc-adequate control (CT; 30 mg/kg); zinc-adequate, diet-restricted (DR; 30 mg/kg); or SZ (300 mg/kg) groups, with and without lead acetate-containing drinking water (200 mg Pb/l) for 3 weeks. Excised femurs were analyzed for bone mineral density (BMD) by dual-energy x-ray absorptiometry, morphometry, and mineral content. MZ had higher femur lead and lower femur zinc concentrations and impaired skeletal growth and mineralization than CT. DR inhibited growth but did not result in higher femur lead concentrations than CT. SZ had higher femur zinc and lower femur lead concentrations than the other treatments. DR and SZ had impaired BMD versus CT and MZ. Lead also retarded skeletal growth and impaired BMD, but an interaction between lead and MZ was only found for femoral knee width, which was lower in MZ exposed to lead. In summary, while MZ deficiency exacerbated bone lead concentration, it generally did not intensify lead toxicity. SZ was protective against bone lead but was detrimental to BMD, suggesting that the optimal level of SZ to reduce lead absorption, while supporting growth and bone development, requires further investigation.
The objective of this pilot study was to investigate the contribution of environmental exposures to lead in the development of cytogenetic damage detected as the frequency of micronuclei (MN) in children. The other aim was to apply the MN assay in combination with fluorescence in situ hybridization (FISH) using a pan-centromeric chromosome probe to elucidate the formation mechanism of induced MN. The examined population was composed of 9-year-old children (n = 92), living in the region where non-ferrous ores are extracted and processed. The non-exposed group consisted of 49 children of the same age from an unexposed recreational area. Exposure to lead was assessed by determination of lead concentrations in blood (PbB) by atomic absorption spectrophotometry, whereas the level of selenium (Se) in serum was detected by using graphite furnace atomic-absorption spectrometry. The frequency of MN was determined by the cytokinesis-block MN assay and fluorescence in situ hybridization performed using a specific pan-centromeric probe. Environmental exposure to lead resulted in significantly increased levels of PbB (5.29 +/- 2.09 versus 3.45 +/- 1.20 microg/dl in controls), although the average level was much below the value of the biological exposure limit = 10 microg/dl. A negative correlation between lead in blood and Se in serum concentrations (P = 0.006) was found for the pooled study population. The results showed a significant difference (P < 0.0001) in the level of MN between the exposed and control group (standard MN test: 2.96 +/- 2.36 versus 1.16 +/- 1.28; FISH technique: 3.57 +/- 3.02 versus 1.43 +/- 1.69, respectively). The frequencies of both centromere-positive (C+MN) and centromere-negative (C-MN) micronuclei were significantly increased in exposed children; however, the contribution of C+MN in the total number of MN in peripheral blood lymphocytes of exposed children was significantly higher than in the controls what may suggest a pro-aneugenic effect of the exposure to lead. The results of multiple regression analysis indicated that the exposure to lead was an important factor affecting the increase in MN frequency what was confirmed by significant correlation between the PbB and MN levels. In conclusion, our results suggest that the exposure to lead may be associated with an increased frequency of MN, especially of C+MN; however, the influence of other factors (e.g. vitamins and minerals in the diet) cannot be excluded.
Chinese hamster cell cultures were treated with different concentrations of lead acetate for a period of one cell cycle. Afterwards chromosome damage and disturbance of the mitotic apparatus were investigated. With increasing lead acetate concentration the mitotic index decreased. However, structural chromosome aberrations, which can be related to true discontinuities of the chromosome structure, and disturbance of the chromosome distribution were never more frequent than in controls. On the other hand, the achromatic lesions of gaps increased with the most concentrated lead acetate solution.
The data concerning the mutagenic, clastogenic and carcinogenic properties of inorganic lead compounds have been conflicting. To investigate whether the genotoxicity of lead is due to indirect effects such as interference with DNA-repair processes, the induction of mutations, sister-chromatid exchanges and strand breaks by lead ions alone as well as in combination with UV light as a standard mutagen were determined. Lead acetate alone does not induce DNA-strand breaks in HeLa cells or mutations at the HPRT locus and sister-chromatid exchanges in V79 Chinese hamster cells. However, at all endpoints tested, lead ions interfere with the processing of UV-induced DNA damage. They inhibit the closing of DNA-strand breaks after UV irradiation and enhance the number of UV-induced mutations and sister-chromatid exchanges, indicating an inhibition of DNA repair. These data point out the necessity to consider such indirect effects when assessing the genotoxicity of metal compounds. As possible mechanisms of repair inhibition we suggest either the interaction with repair enzymes such as polymerase or ligase or else the interaction with calcium-regulated processes, for example with calmodulin.
We have investigated the activity of insoluble and soluble lead compounds in inducing mutagenesis, cell transformation and sister chromatid exchange in mammalian cells. Insoluble lead sulfide, readily phagocytized, was more than four times as toxic to V79 cells on a microM basis, than two moderately soluble lead compounds although the exposure time for the soluble salts was five times longer. These findings demonstrate the importance of different cellular mechanism(s) of metal uptake and bioavailability. Both insoluble lead sulfide and more soluble lead nitrate were mutagenic at the HPRT locus in V79 cells. Although less mutagenic at the higher concentrations, lead nitrate at a concentration of 500 microM enhanced the mutation frequency greater than 6-fold above background following a 5-day exposure. Although the mechanism(s) by which lead induces mutations is unknown, failure of both compounds to induce SCE and DNA single-strand breaks, detectable by alkaline elution, suggests that lead-induced mutations may not be a result of direct damage to DNA but may occur via indirect mechanisms including disturbances in enzyme functions important in DNA synthesis and/or repair, or in DNA-helical structure. Lead acetate also transformed SHE cells in a dose-response fashion following a 48-h exposure. Our results indicate that lead compounds may be genotoxic by an indirect mechanism, and lend support to the view that lead is a carcinogen.
The inducibility of chromosome aberrations and micronuclei by lead acetate was examined in vivo in bone marrow cells of rats after its single or repeated intraperitoneal administration. Lead acetate caused a small but significant increase in the frequencies of chromosomal aberrations (mostly chromatid gaps) and micronuclei. Chromatid breaks and acentric fragments were less frequent in occurrence. Complex aberrations such as chromatid exchanges and multiple breaks were never observed. Our findings suggest that lead acetate exerts a weak clastogenicity on rat bone marrow cells.
The influence of lead on sperm morphology, sister chromatid exchanges or on micronuclei formation was studied on male rabbits after exposure to doses of 0, 0.25, and 0.50 mg lead acetate/kg body weight subcutaneously injected three times a week during 14 weeks, each on a group of five rabbits. At the end of exposure phase the lead in blood concentrations of the three groups of rabbits were 0.32, 2.57, and 2.97 mumol/l respectively. The results did not show any evidence of treatment related effects on sperm count or on morphologic abnormalities of the sperms, neither on the histopathology of the testis. Statistical analysis of the number of sister chromatid exchanges per metaphase in lymphocytes indicated no differences between the groups. Also no dose dependent effect was observed on the relative number of micronuclei in bone marrow erythrocytes. The different susceptibility to lead in different organ systems of the rabbits was discussed.
Lead has been tested for genotoxic potential in a range of mutagenicity assays. Such studies report varying effects, and the results are equivocal. The reasons for this may be related to problems of the solubility of many lead compounds, their inability to dissolve in biological fluids, chemical interferences resulting in conflicting observations, the nonspecificity of the assays used, the delivery of toxic doses to specific genetic processes or the mediation of genotoxicity through indirect mechanisms. Occupational and environmental exposures to lead have also been associated with increases in chromosomal damage in humans at moderate to high exposures, although this evidence is contradictory. Where positive findings have been reported, many are related to exposure. The possibility that lead is genotoxic is by no means clear, but evidence is still accumulating.
Cadmium and lead have been shown to induce cellular transformations and gene mutations in cultured rodent cells, as well as tumours in live animals. However, the mechanisms by which these metals cause cellular transformations and mutations in human cells have not been explored. In this study, we investigated the abilities of cadmium and lead to induce anchorage-independent transformations and hprt gene mutations in diploid human fibroblasts. Human fibroblasts were exposed to either cadmium acetate (0-60 microM) or lead acetate (0-2 mM) for 24 h. After removal of the metals, the cells were kept in exponential growth for 7 and 9 days before mutation and anchorage-independence assays were taken, respectively. Both cadmium and lead significantly induced anchorage-independent colonies in dose-dependent manners; the frequencies of anchorage-independent colonies induced by these metals were similar to those induced by N-methyl-N'-nitro-N-nitrosoguanidine at approximately equal cytotoxic dose ranges (30-10% survival). 3-Aminotriazole at non-cytotoxic dosages decreased catalase activity by >80%, and markedly enhanced cadmium-induced cytotoxicity and anchorage-independent colonies. Cadmium uptake by human fibroblasts was not affected by 3-aminotriazole co-administered with 10 microM of cadmium; whereas cadmium uptake and accumulation were enhanced 1.5-fold by 3-aminotriazole co-administered with 1-2.5 microM of cadmium. Lead-induced anchorage-independence or cytotoxicity was not affected by 3-aminotriazole co-treatment; however, 3-aminotriazole did significantly enhance lead uptake and accumulation in human fibroblasts. Neither cadmium- nor lead-induced 6-thioguanine-resistant mutation frequency in human fibroblasts. Co-administering these metals with 3-aminotriazole did not enhance mutations in human fibroblasts. These results suggest that cadmium and lead may both act as tumour promoters in diploid human fibroblasts, and that reactive oxygen species is more important in cadmium- than lead-induced cytotoxicity and anchorage-independence.
This article reviews the effects of lead on genetic systems in the context of lead's various other toxic effects and its abundance and distribution in the environment. Lead is perhaps the longest used and best recognized toxic environmental chemical, yet it continued be used recklessly until only very recently. Lead is thus a lesson in the limitations and strengths of science, human conscience and common sense. Lead has been tested and found to be capable of eliciting a positive response in an extraordinarily wide range of biological and biochemical tests; among them tests for enzyme inhibition, fidelity of DNA synthesis, mutation, chromosome aberrations, cancer and birth defects. It reacts or complexes with many biomolecules and adversely affects the reproductive, nervous, gastrointestinal, immune, renal, cardiovascular, skeletal, muscular and hematopoietic systems as well as developmental processes. It is likely that lead is a selective agent that continues to act on and influence the genetic structure and future evolution of exposed plant and animal populations.
We have studied the in vitro effects of lead (Pb) as Pb-acetate (0. 1-1000 ppm) on the activation of rat spleen (SP) cells. At a concentration of 0.5 to 200 ppm, Pb augmented the uptake of [3H]thymidine, progression of SP cells through the cell cycle, and allogeneic and syngeneic mixed lymphocyte reactions. However, at concentrations above 200 ppm, Pb inhibited the proliferation of these cells. To understand the cellular and molecular basis of these responses, we examined the effects of Pb on the proliferation of isolated T and/or B cell populations. Pb failed to stimulate the proliferation of isolated T and B cells; however, the addition of gamma-irradiated B cells to T cell cultures or irradiated T cells to B cell cultures resulted in Pb-induced incorporation of [3H]thymidine. On the other hand, macrophages were unable to reconstitute this response. Pb also induced a significant rise in the intracellular concentration of inositol 1,4,5-trisphosphate in SP cells; however, unlike the activation of lymphocytes through the antigen receptors, Pb did not significantly stimulate protein tyrosine kinase activity. These observations suggest that Pb facilitates the T cell-B cell interaction-dependent proliferation of lymphocytes through a signaling pathway(s) independent of the antigen receptor.
Oxidative damage associated with the presence of lead (Pb) in the brain has been proposed as one possible mechanism involved in Pb toxicity. To investigate this hypothesis, we examined the long-term effects of Pb2+ on parameters of oxidative stress in the brain from rats chronically exposed to the metal (1 g Pb acetate/1 drinking water). After 8 weeks of treatment, Pb2(+)-intoxicated rats (blood Pb concentration > 100 microg/dl) showed lower body weight, and lower hematocrit and 5-aminolevulinic acid dehydratase activity as compared to controls. The content of brain 2-thiobarbituric acid-reactive substances (TBARS), an indicator of lipid oxidation, was significantly (P < 0.05) higher in the Pb2(+)-intoxicated animals than in controls. Higher activities of the antioxidant enzymes glutathione reductase and glutathione peroxidase, and a lower (44%) level of ubiquinol 10 were found in the brain of the Pb2(+)-treated rats, compared to controls. A negative correlation between brain ubiquinol 9 (r2 = 0.79), 10 (r2 = 0.84) and blood Pb concentration was observed. Brain alpha-tocopherol levels, superoxide dismutase activity and parameters of oxidative damage to proteins were similar between control and Pb2(+)-treated rats. The present results indicate that chronic Pb2+ intoxication induces an oxidative stress situation in rat brain.
The aim of the present study was to investigate the possible genotoxic effects, exerted by the pyrethroid cypermethrin and by either of the metals cadmium and lead alone or in combination, on bone marrow cell chromosomes in a subchronic experiment. Outbred male Wistar rats were treated per os for 4 weeks in a five-time per week schedule with 5.54, 11.08, or 22.16 mg/kg cypermethrin (1/100, 1/50, 1/25 LD50) alone, or in combination of 1/100 and 1/50 LD50 cypermethrin with 2.0 mg/kg cadmium chloride or 10 mg/kg lead acetate. On the day following the last treatment, the animals were sacrificed and bone marrow from the femur was prepared. Twenty metaphases from 10 animals per group were evaluated. The evaluation comprised the frequency of aberrant cells, the numerical and structural aberrations, and the alterations in relative organ weights. In the dosage used, cypermethrin and cadmium alone caused no significant increase in the chromosomal aberrations, and lead acetate caused an increase of the numerical aberrations only. Combination of cypermethrin and cadmium also failed to induce significant chromosomal effects. The cypermethrin + lead combination, however, induced a significant increase of structural chromosomal aberrations, predominantly of all acentric fragments. This lead to the conclusion that the simultaneous administration of lead and cypermethrin results in an enhanced genotoxic effect.
Lead poisoning is a worldwide, environmental health-hazard that affects children and adults. In this review we discuss the effects of lead on gene expression due to both general and specific mechanisms. In particular we focus on the ability of lead to substitute for biologically essential metals such as calcium and zinc in metal-binding domains of cytoplasmic enzymes, nuclear transcription factors and other proteins. The binding of lead to these proteins causes an alteration of their activity resulting in aberrant expression of their own genes and in some cases their target genes. Finally, we discuss the impact of microarray technology on the study of the genome-wide effects of lead and other toxicants on gene expression.
The protective effect of calcium given orally by gavage with two doses (40 and 80mg/kg body weight) was evaluated against clastogenecity induced by lead acetate with two concentrations (200 and 400mg/kg diet) on bone marrow and spermatocyte cells of mice in vivo. The parameter screened was percentage of chromosomal aberrations with and without gaps and sperm abnormalities. Statistical analyses indicated the protection efficacy of calcium with the high dose rather than the other in both types of mouse cells. The observation from the laboratory tests, dealing that lead acetate can be considered as an environmental genotoxic material. We recommended that it must be administered of calcium (as calcium chloride) as a protective agent to reduce the genotoxic effect of lead in the somatic and germ cells.
The underlying mechanisms by which lead ions produce their deleterious effects prior to the onset of clinical symptoms are incompletely understood. This study aimed to assess lead-induced cell toxicity mechanisms by focusing on the effects of the metal on cell growth, DNA synthesis, cellular ATP, intracellular hexosaminidase activity and lysosomal function, and examine the possible cytoprotective role of fetal calf serum (FCS). Several human dermal cultured fibroblast lines were exposed to Pb (400 microM) for 1-6 days with 2, 5, and 10% FCS. The earliest toxic effect of Pb was significant inhibition of DNA synthesis after 24 h direct exposure; this harmful effect was not progressive during the first 3 days, but worsened clearly on the 4th day regardless of the FCS concentration. Atime-dependent depletion of intracellularATP content was also caused by ionic lead, thereby compromising the cell energy charge which precedes cell death. Fibroblast growth was progressively and significantly inhibited from day 2 onwards; the greatest noxious effect was observed in the presence of 2% FCS: 49% reduction in cell proliferation after 5 days. Lead salts produced loss of cell adhesion to the culture dish which worsened from the 2nd day and was more pronounced when FCS in growth medium was decreased. Toxic actions on lysosomal membrane integrity provoked a decrease in neutral red uptake (NRU) which was exposure time-dependent and more marked with 2% FCS. In contrast, increased relative NRU (to 20% at 4 days), suggestive of endocytosis-induced lysosome enlargement, was observed in Pb-exposed cells. Intracellular hexosaminidase activity was not negatively affected until 5 days after exposure to Pb salts. FCS had a significant cytoprotective effect on Pb-induced toxicity.
Lead is present in the natural and occupational environment and is reported to interact with DNA, but the mechanism of this interaction is not fully understood. Using the alkaline comet assay we showed that lead acetate at 1-100μM induced DNA damage in isolated human lymphocytes measured the change in the comet tail length. At 1 and 10μM we observed an increase in the tail length, whereas at 100μM a decrease was seen. The former effect could follow from the induction of DNA strand breaks and/or alkali-labile sites (ALS), the latter from the formation of DNA-DNA and/or DNA-protein cross-links. No difference was observed between tail length for the alkaline and pH 12.1 versions of the assay, which indicates that strand breaks and not ALS are responsible for the tail length increase induced by lead. The neutral version of the test revealed that lead acetate induced DNA double-strand breaks at all concentrations tested. The presence of spin traps, 5,5-dimethyl-pyrroline N-oxide (DMPO) and N-tert-butyl-α-phenylnitrone (PBN) did not influence the level of DNA damage induced by lead. Post-treatment of the lead-damaged DNA (at 100μM treatment concentration) by endonuclease III (Endo III) and formamidopyrimidine-DNA glycosylase (Fpg), enzymes recognizing oxidized DNA bases, as well as 3-methyladenine-DNA glycosylase II, an enzyme recognizing alkylated bases, gave rise to a significant increase in the extent of DNA damage. Proteinase K caused an increase in comet tail length, suggesting that lead acetate might cross-link DNA with nuclear proteins. Vitamin A, E, C, calcium chloride and zinc chloride acted synergistically on DNA damage evoked by lead. The results obtained suggest that lead acetate may induce single-strand breaks (SSB) and double-strand breaks (DSB) in DNA as well as DNA-protein cross-links. The participation of free radicals in DNA-damaging potential of lead is not important and it concerns other reactive species than could be trapped by DMPO or PBN.
Interactions of chemicals with the microtubular network of cells may lead to genotoxicity. Micronuclei (MN) might be caused by interaction of metals with tubulin and/or kinesin. The genotoxic effects of inorganic lead and mercury salts were studied using the MN assay and the CREST analysis in V79 Chinese hamster fibroblasts. Effects on the functional activity of motor protein systems were examined by measurement of tubulin assembly and kinesin-driven motility. Lead and mercury salts induced MN dose-dependently. The no-effect-concentration for MN induction was 1.1 microM PbCl(2), 0.05 microM Pb(OAc)(2) and 0.01 microM HgCl(2). The in vitro results obtained for PbCl(2) correspond to reported MN induction in workers occupationally exposed to lead, starting at 1.2 microM Hg(II) (Vaglenov et al., 2001, Environ. Health Perspect. 109, 295-298). The CREST Analysis indicate aneugenic effects of Pb(II) and aneugenic and additionally clastogenic effects of Hg(II). Lead (chloride, acetate, and nitrate) and mercury (chloride and nitrate) interfered dose-dependently with tubulin assembly in vitro. The no-effect-concentration for lead salts in this assay was 10 microM. Inhibition of tubulin assembly by mercury started at 2 microM. The gliding velocity of microtubules along immobilised kinesin molecules was affected by 25 microM Pb(NO(3))(2) and 0.1 microM HgCl(2) in a dose-dependent manner. Our data support the hypothesis that lead and mercury genotoxicity may result, at least in part, via disturbance of chromosome segregation via interaction with cytoskeletal proteins.
In this study, chromosomal damage induced in vitro by lead acetate in human melanoma cells (B-Mel) was evaluated using the cytokinesis-blocked micronucleus assay and sister chromatid exchange (SCE) analysis. Lead acetate (10-6, 10-5 and 10-3 mM) induced micronuclei and SCE formation in a dose-dependent manner. Treated cells showed a decrease in cell viability but not concomitant cell death by apoptosis (lead acetate failed to induce internucleosomal DNA fragmentation at any of the doses tested). One important observation emerging from this study was that low-level lead exposure in vitro is able to induce significant cytogenetic damage in human melanoma cells, indicating an increased sensitivity of B-Mel cells to lead acetate.
The protective action of vitamins C and E against lead acetate-induced reduced sperm count and sperm abnormalities in Swiss mice has been studied. Intraperitoneal injection of lead acetate (10mg/kg body weight) in the present study stimulates lipid peroxidation in the testicular tissue, indicated by a significant increase in malondialdehyde content in the experimental mice group. This is associated with an increased generation of noxious reactive oxygen species (ROS). Significantly reduced sperm count associated with increased sperm abnormality percentage in the lead-injected mice group compared to controls substantially proves the ongoing damaging effects of lead-induced ROS on developing germ cells. However, intraperitoneal administration of vitamin C (Vit C) at a concentration equivalent to the human therapeutic dose (10 mg/kg body weight) was able to minimize significantly the testicular malondialdehyde content with a concomitant increase in sperm count and significant decrease in the percentage of abnormal sperm population. Vitamin E (Vit E) (100 mg/kg body weight) treatment of a batch of lead-injected mice had a similar effect as Vit C but with a comparatively lower efficacy. On the other hand, coadministration of both vitamins (Vit C + Vit E) at the above mentioned doses to lead-treated mice led to the most significant decline in malondialdehyde content along with elevated sperm count and reduction in the percentage of abnormal sperm population. The protective action and the synergistic action of both vitamins (C and E) against lead-induced genotoxicity are discussed.
Lead compounds are known genotoxicants, principally affecting the integrity of chromosomes. Lead chloride and lead acetate induced concentration-dependent increases in micronucleus frequency in V79 cells, starting at 1.1 microM lead chloride and 0.05 microM lead acetate. The difference between the lead salts, which was expected based on their relative abilities to form complex acetato-cations, was confirmed in an independent experiment. CREST analyses of the micronuclei verified that lead chloride and acetate were predominantly aneugenic (CREST-positive response), which was consistent with the morphology of the micronuclei (larger micronuclei, compared with micronuclei induced by a clastogenic mechanism). The effects of high concentrations of lead salts on the microtubule network of V79 cells were also examined using immunofluorescence staining. The dose effects of these responses were consistent with the cytotoxicity of lead(II), as visualized in the neutral-red uptake assay. In a cell-free system, 20-60 microM lead salts inhibited tubulin assembly dose-dependently. The no-observed-effect concentration of lead(II) in this assay was 10 microM. This inhibitory effect was interpreted as a shift of the assembly/disassembly steady-state toward disassembly, e.g., by reducing the concentration of assembly-competent tubulin dimers. The effects of lead salts on microtubule-associated motor-protein functions were studied using a kinesin-gliding assay that mimics intracellular transport processes in vitro by quantifying the movement of paclitaxel-stabilized microtubules across a kinesin-coated glass surface. There was a dose-dependent effect of lead nitrate on microtubule motility. Lead nitrate affected the gliding velocities of microtubules starting at concentrations above 10 microM and reached half-maximal inhibition of motility at about 50 microM. The processes reported here point to relevant interactions of lead with tubulin and kinesin at low dose levels.
The content of cadmium and lead, as risk factors of environment, in liver and kidneys of wild animals as brown hare (Lepus europaeus), yellow-necked mouse (Apodemus flavicollis), wood mouse (Cleithrionomys glareolus), and red deer (Cervus elaphus) were studied. Samples were analyzed by the atomic absorption spectrophotometry method (AAS). The highest levels of cadmium were found in kidneys (0.213-2.387 mg/kg) of all animal species. The concentration of cadmium in liver was 0.032-0.258 mg/kg. The analysis of lead showed that the concentration of this element was higher in kidneys of yellow-necked mouse and wood mouse (0.503-0.780 mg/kg) than in liver (0.177-0.268 mg/kg). In brown hare and red deer a higher accumulation of lead in liver (0.221-1.904 mg/kg) in comparison with kidneys (0.115-0.561 mg/kg) is reported.
Natural dietary antioxidants are extensively studied for their ability to protect cells from miscellaneous damages. Origanum majorana L., Lamiaceae, is a potent antioxidant. The effect of administration of O. majorana (volatile oil, alcoholic and aqueous extracts) on oral administration of lead acetate in the diet of mice at concentration 0.5% (W/W) for one month were studied by measuring serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), urea and creatinine, histopathological changes of the liver and kidney and genotoxicity including, rate of micronucleus and chromosomal aberrations in bone marrow cells. Mice were treated with the 3 different forms of O. majorana, one month before and maintained with lead acetate administration. The 3 forms of O. majorana induced a significant decrease in serum activities of transaminases (AST & ALT), ALP, urea and creatinine and improved the liver and kidney histology in comparison with lead acetate treated group. Alcoholic extracts of O. majorana significantly reduced the rate of micronucleus, number of aberrant cells and different kinds of chromosomal aberrations. Volatile oil extract significantly reduced the rate of micronucleus and chromosomal fragments. Aqueous extract and volatile oil also of O. majorana significantly reduced number of gaps, ring chromosome and stickiness. It could be concluded that O. majorana plays an important role in ameliorating liver and kidney functions and genotoxicity induced by lead toxicity.
The effects of lead acetate in the diet (0.5% w/w) on reduced GSH, activity of phase II metabolizing enzyme glutathione S-transferase (GST), lipid peroxidation in liver homogenate and bone marrow chromosomes of mice simultaneously supplemented with powdered turmeric and myrrh for 8 weeks were investigated. Five groups of Swiss male albino mice, each of 30 mice, the first group received a basal diet and served as negative control, the second group received basal diet supplemented with lead acetate only and served as positive control. The other three groups received basal diet supplemented with lead acetate and 1% or 5% turmeric powder and 1% myrrh powder, respectively. Results revealed a significant decrease in the amount of GSH in all treated groups compared with negative control. Also, the activity of GSH S-transferase was significantly decreased in positive control compared with other groups. However, co-administration of the protective plants resulted in a significant increase in the activity of GST compared with both positive and negative control groups. Furthermore, lipid peroxidation was significantly increased in positive control alone, while co-treatment with the protective plants resulted in reduction in the level of lipid peroxidation by 31% and 49% in mice receiving 1% and 5% turmeric powder respectively and 45% in 1% myrrh treated when compared with their respective positive control group. Lead genotoxicity was confirmed through significant reduction in the number of dividing cells, increased total number of aberrant cells and increased frequency of chromosomal aberrations. Simultaneous treatment with these plants significantly reduced the genotoxicity induced by lead administration and the powerful protection was observed with 5% powdered turmeric. It may be concluded that turmeric and myrrh are useful herbal remedies, especially for controlling oxidative damages and genotoxicity induced by lead acetate intoxication.
To investigate the effects of lead exposure to rat placenta and pups during different gestation periods.
All 108 Wistar rats (72 females, 36 males) were randomly divided into four groups. All rats were orally fed with 0.025% lead acetate during different gestation periods. Blood was obtained from the abdominal vena cava and the lead level in maternal blood was measured by means of atomic absorption spectrometry at the end of the pregnancy. The number of pups, their body weight, body length and tail length were measured. The effects of lead to rat placenta were observed by level of microscopy, optical microscopy and electronic microscopy.
Experimental groups the blood lead level at the end of gestation were above 0.483 micromol/L. There were significant differences among, of pups, during different groups (P < 0.01). Among them the drinking lead group of whole distant was the lowest in placenta weight [(0.31 +/- 0.13) g] body weight of pups [(2.08 +/- 0.88) g] length and tail length of pups [(2.37 +/- 0.32) cm, (0.98 +/- 0.09) cm]. There were significantly differences between the experimental groups and controls. Maternal blood lead level was negatively related to placenta weight (r = 0.652, P < 0.01), and had no relation with the body weight of pups (r = -0.107, P = 0.46). In the experimental groups of lead poisoned rats, the placenta showed focus necrosis in the deciduas, and increased the trophoblastic giant cells and light staining cells in the trophospongium. Trophoblast in the labyrinth and trophospongium showed degeneration; fibrin deposition around the villi was increased. Microvilli around the trophoblast were shorter and less, mitochondrion was swollen and decreased in number, rough endoplasmic reticulum was distended and ribosomal number on membrane decreased.
Lead exposure during different gestation periods should have a traumatic effect on the trophoblast, leading to interference of nutrition and oxygen exchange. Furthermore, the blood supply to the placenta and nutrition and oxygen exchange between mother and pups were also interfered, leading to reduction of placenta weight and retardation of development of pups.
No abstract available.
The nucleolus is a compulsory nuclear structure of all cells of eukaryotes. The quantitative and qualitative characteristics of nuclei show the functional activity of a cell, the rate of its synthesis of RNA and portents, and its metabolic state. Heavy metals (zinc chloride and lead acetate) were comparatively investigated for their effects on the nucleolar apparatus of bone marrow cells in in vivo experiments. Zinc chloride and lead acetate were ascertained to damage the nucleolar apparatus of cells, thus decreasing their transcriptional activity or irreversibly damaging them.
In order to investigate the effects of Zn on Pb toxicities. Proportion of abnormal sperm, percentage of micronucleated polychromatic erythrocyte (MPCE), serum thyroid hormones (T(3), T(4)) and cortisol were measured. Rats received intraperitoneal injection of 25 mg/kg Pb acetate, 4 mg/kg Zn acetate, both Pb acetate and Zn acetate, or normal saline as controls, once every two days, 7 times in total. No significant differences in whole blood Pb were detected between groups received Pb alone or both Pb and Zn. On the contrary, the concentration of whole blood Zn in the group given Zn alone was significantly higher than that in the group that received both Pb and Zn. In the groups given Pb alone or both Pb and Zn, proportion of abnormal sperm, frequency of MPCE and serum cortisol were significantly higher than those in controls, whereas serum T(3) and T(4) were significantly lower than in controls. In the group given both Pb and Zn, T(4) was decreased most obviously among the four groups. While the proportion of abnormal sperm was less in the group given both Pb and Zn than in the group given Pb alone. These findings suggest that Zn coadministration might alleviate toxic effects of Pb on the male reproductive system, whereas it could enhance the toxicity on thyroid function. Zn did not affect the toxicities of Pb on cytogenetic systems as indicated by MPCE percentage, and on serum cortisol levels under the dose of the present study. Our results suggested the double-edged effects of Zn on Pb toxicities in different organs. Therefore, the effects of Zn on Pb toxicities should be evaluated systematically.
The genotoxic effects of mercury chloride and lead acetate were evaluated in vivo using the micronucleus (MN) assay on acridine-orange (AO) stained peripheral blood erythrocytes, gill and fin epithelial cells of Carassius auratus auratus. Fish were exposed to three different concentrations of mercury chloride (MC) (1 microg/, 5 microg/L and 10 microg/L) and lead acetate (LA) (10 microg/L, 50 microg/L and 100 microg/L) for 2, 4 and 6 days. A single dose of 5 mg/L cyclophosphamide was used as a positive control. In addition to micronuclei, nuclear buds (NBs) were assessed in the erythrocytes. The ratio of polychromatic and normochromatic erythrocytes (PCE/NCE) in peripheral blood was also evaluated to assess cytotoxicity. MN frequencies in all three tissues were elevated in fish exposed to both LA and MC. However, NBs showed different sensitivity to metal treatments. MN frequencies in both control and treated fish were highest in gill cells and generally lower in erythrocytes and fin cells. PCE/NCE rations decreased in relation to MC and LA treatments. The results of this study indicate that LA and MC have genotoxic and cytotoxic damage in fish and confirmed that AO staining is a suitable technique for in vivo MN test in fish.
Oxidative stress is considered as a possible molecular mechanism involved in lead toxicity. This study was carried out to investigate whether lead acetate could induce oxidative stress in mice, and the following damages as well. Lead acetate was given orally to mice for 4 weeks at doses of 0, 10, 50, 100mg/kg body weight every other day, respectively. Production of reactive oxygen species (ROS) and malondialdehyde (MDA) were measured as indicators of oxidative stress. DNA damage in peripheral blood lymphocytes was determined by comet assay. Ultrastructure alteration was detected using transmission electron microscopy. The alterations of p53, Bax, and Bcl-2 expression were determined by western blotting. The results showed that lead acetate significantly increased the levels of ROS and MDA in mice. Meanwhile, severe DNA damage and ultrastructure alterations were obviously observed. In addition, p53 and Bax expressions increased and the imbalance of Bax/Bcl-2 occurred. Therefore, it strongly suggests that lead may induce oxidative stress and change the expressions of apoptosis-related proteins in mouse liver.
L (1995) Study of lead acetate genotoxicity using the micronucleus test in mice
- C Rusov
- R Živković
- S Popović
- Joji Ćmaličević
Rusov, C, Živković, R, Popović, S, Joji Ćmaličević, L (1995) Study of lead acetate genotoxicity using the micronucleus test in mice. Acta Vet 45: 317–322.