Influence of selenium supplementation during chelation of lead in rats.

The influence of selenium supplementation during chelation therapy to reduce body burden and toxicity of lead was investigated in rats. Selenium had marginal effects on liver, kidney and blood lead decorporation by calcium disodium ethylenediamine tetra acetic acid (CaNa2EDTA) and activation of inhibited delta- aminolevulinic acid dehydratase (ALAD) activity by calcium trisodium diethylenetriamine penta acetic acid (CaNa3DTPA). Selenium supplementation however, had no influence on lead induced inhibition of renal and hepatic transaminases and alkaline phosphatase. The results suggest that selenium supplementation slightly augments lead mobilization by chelating drugs.     

Beneficial effects of zinc supplementation during chelation treatment of lead intoxication in rats

The ability of zinc to enhance the efficacy of commonly used chelating drugs in lead intoxication and to reduce the resulting zinc imbalance, was investigated in rats. The simultaneous zinc supplementation increased urinary lead elimination by calcium disodium ethylenediaminetetraacetate (Ca disodium EDTA) and 2,3 dimercaptosuccinic acid (DMSA). Combination therapy was also effective in potentiating the depletion of blood, hepatic and renal lead by calcium disodium EDTA and D-penicillamine (DPA), renal lead by DMSA and reversal of inhibited blood delta-aminolevulinic acid dehydratase (ALAD) activity by calcium disodium EDTA and DPA. The body zinc status was also maintained as reflected by urinary, blood and tissue levels of zinc.     

Influence of chelation therapy on acute lead intoxication in rats

The intraperitoneal administration of Pb acetate (5 × 20 mg Pb/kg per day) evokes a moderate and transient hypochromic anemia, a long-lasting enhanced urinary excretion of -aminolevulinic acid whereas the urinary excretion of alkaline phosphatase is not affected and that of lactic debyhrogenase only marginally. It is concluded that neither the hematologic response nor the slight nephrotoxicity are responsible for the lethal action of Pb. Chelate treatment started 3 days after the last Pb dose and was continued over 7 weeks. The daily intraperitoneal dose was 25, 50, and 100 mol/kg, respectively. The efficacy in promoting the urinary excretion of Pb decreased in the following order: Ca diethylenetriaminepentaacetate > 2,3-dimercaptopropane-1-sulfonate > Zn diethylenetriaminepentaacetate > D-penicillamine. This effect was mainly due to the mobilization of skeletal Pb. The chelating agents also lowered the excretion of -aminolevulinic acid but failed to exert a beneficial influence on the anemia and the lethal action of Pb. These negative results raise questions about the usefulness of chelation therapy in cases of acute Pb poisoning.     

Influence of dietary zinc on lead toxicity in rats

The influence of dietary zinc (5 and 50 μg/ml of drinking water) on the toxicity of lead (100 μg/ml) was studied in rats fed a semipurified diet for 140 days. Results indicated that high dietary zinc reduced the lead levels in various tissues. The antagonism between lead and zinc was also manifested by reduction of zinc levels in plasma, liver and tibia of the lead-exposed animals. Lead exposure, as expected, produced an elevation of urinary δ-aminolevulinic acid (U-ALA) throughout the experiment. This increase, however, was greater in the animals receiving low zinc. Erythrocyte zinc protoporphyrin (ZPP) was not only affected by lead, but also by dietary zinc and by the age of the rats. As expected, linear regression analysis of the data showed that RBC-Pb correlated highly with δ -aminolevulinic acid dehydratase (ALA-D), ZPP and U-ALA; moreover, plasma zinc also correlated with RBC-Pb and ZPP.     

Essentiality, toxicology and chelation therapy of zinc and copper

Both zinc and copper are essential minerals that are required for various cellular functions. Although these metals are essential, they can be toxic at excess amounts, especially in certain genetic disorders. Zinc and copper homeostasis results from a coordinated regulation by different proteins involved in uptake, excretion and intracellular storage/trafficking of these metals. Apart from zinc transporters (ZnT) families and Cu-ATPase, metallothionein is an important storage protein for zinc and copper. Metallothioneins are intracellular polypeptides with a remarkable ability to bind metallic ions. These proteins bind both essential metals indispensable for the organism and also toxic metals (e.g. cadmium or lead). Metallothioneins play a critical role to maintain zinc and copper homeostasis. In this review, we summarize the toxicity of zinc and copper and the potential treatment for zinc or copper toxicity by zinc- or copper-specific chelators as well as strategy to up-regulate metallothionein.     

Combined chelation therapy in reducing tissue lead concentrations in suckling rats.

The very young are more prone to lead poisoning than adults, and the treatment with chelating agents, either as monotherapy or combined treatment, is still a matter of dispute. The purpose of this work was to evaluate the efficiency of three chelating agents administered either as monotherapies or as combined treatments in sucklings. Lead acetate (5 mg Pb kg(-1) i.p.) was administered to the 7-day-old rat pups in eight litters on experimental day 1 and chelating agents on experimental days 2 and 3. Pups were divided into six groups: (1) untreated control; (2) EDTA (calcium disodium ethylendiaminetetraacetate, 0.3 mmol kg(-1) i.p. at 4 p.m.); (3) meso-DMSA (meso-2,3-dimeracaptosuccinic acid, 0.5 mmol kg(-1) p.o. at 10 a.m.); (4) rac-DMSA (racemic-2,3-dimeracaptosuccinic acid, 0.5 mmol kg(-1) p.o. at 10 a.m.); (5) EDTA+meso-DMSA; and (6) EDTA+rac-DMSA. Rats were killed on experimental day 5. Tissue element concentrations were analyzed by atomic absorption spectrometry. Treatment with EDTA did not affect tissue Pb, but it reduced Zn in the carcass and liver. Meso-DMSA reduced Pb in the kidneys and brain, and it did not affect organ essential elements. Rac-DMSA most efficiently reduced Pb concentrations in the carcass, kidneys and brain, but it also reduced Zn and Cu in the liver and Zn in the kidneys. Combined treatments with EDTA never improved the efficiency of either DMSA isoform in decreasing tissue Pb but they did reduce tissue Zn concentrations. All treatments caused the same decrease in the carcass Ca concentrations. The results do not support combined treatment in this age group, which is especially sensitive to trace element deficiencies, and suggest that meso-DMSA might be the treatment of choice in acute lead poisoning in infants.     

Stable isotopic tracers of lead mobilized by DMSA chelation in low lead-exposed rats.

The ability of DMSA to mobilize skeletal lead or effect a redistribution of endogenous lead to other target organs in low lead-exposed organisms is unclear. Discrepant results of past studies of DMSA and other lead chelators (e.g., CaNa2EDTA) may be due, in part, to experimental differences and difficulties in distinguishing mobilized skeletal lead from other endogenous or exogenous lead sources. Therefore, the influence of DMSA on the mobilization and redistribution of lead in skeletal and soft tissue compartments of low lead-exposed female Wistar (115-125 g) rats was investigated using ultraclean stable lead isotope tracer techniques. Rats that had been reared on a low lead-level diet (lead intake approximately 80 ng Pb/g body/day) were fed 206Pb-enriched drinking water (210 ng Pb/ml) for 1.5 days and then were chelated with a single ip injection of a 0.11 mmol/kg dose of DMSA. Blood, kidney, brain, tibia, urine and feces were collected 24 hr after chelation and analyzed for lead concentrations by graphite furnace atomic absorption spectrometry and for lead isotopic compositions by thermal ionization mass spectrometry. These analyses demonstrated that DMSA chelation significantly increased (15-fold) the diuresis of labile soft tissue lead, but not skeletal lead. DMSA also appeared to effect a redistribution and input of a comparable amount of lead to the skeleton and smaller relative amounts of lead to the soft tissues (blood, kidney) of the chelated animals. The clinical significance of these latter observations beyond the context of this preliminary study is not clear.     

Changes in zinc, copper and selenium status during adjuvant-induced arthritis in rats

Trace elements such as zinc, copper and selenium are involved in the pathogenesis of inflammatory diseases. In order to obtain more information about the overall movements of these minerals during the evolution of an experimental chronic inflammatory process, trace element levels were determined in five body compartments of the rat at several time intervals after induction of adjuvant arthritis. Rapid and significant changes in plasma zinc and copper levels and in liver zinc levels were observed. These modifications occurred as early as those in biochemical parameters of inflammation such as serum fibrinogen and ceruloplasmin, and preceded the appearance of any clinical symptom of the disease. Inverse correlations were found between plasma zinc levels and these two biochemical indices. Other modifications in trace element levels were observed two weeks after disease induction, the most important being a considerable increase in liver copper levels. Although food intake of affected animals decreased with the progression of the disease, there was no evidence of depletion in zinc and copper levels over the study period. A redistribution of body zinc between different biological compartments (mainly plasma and liver) occurred simultaneously with an accumulation of copper in several organs. The decreasing selenium status of animals was not clearly related to the inflammatory process.     

Calcium supplementation efficiently reduces lead absorption in suckling rats

The effect of calcium supplementation on tissue lead was evaluated in suckling Wistar rats. Such data are not yet available in the literature. The following artificial feeding regimen was used for calcium supplementation: cow's milk by addition of 1%, 3% or 6% Ca as CaHPO(4)x2H(2)O suspension to increase the daily calcium intake about 1.4, 2 or 3 times above control values. Artificial feeding was applied during 7 hr each day for nine consecutive days (from day 6 through 15 after birth). The effect of such treatment on lead absorption and elimination was evaluated in two separate experiments: calcium supplementation during oral lead exposure (as acetate; daily dose 2 mg Pb/kg body wt.; total Pb dose 18 mg/kg body wt.) or after a single intraperitoneal lead administration (5 mg/kg body wt.). At the end of experiments, lead in tissues (liver, kidneys, brain and carcass), and essential elements (Ca, Fe, Zn, Cu) were analysed by atomic absorption spectrometry. Calcium supplementation caused a statistically significant decrease of lead in all tissues of sucklings orally exposed to lead. This decrease was dose-related being about 1.3, 1.5 and 2 times lower in groups supplemented with 1%, 3%, or 6% calcium compared to controls, respectively. Increased calcium intake had no effect on incorporated lead after parenteral lead exposure. Calcium supplementation increased carcass calcium and had no effect on trace elements in tissues, pups' general appearance and body weight gain. It is concluded that higher calcium intake might be a way of efficient reduction of lead absorption during the suckling period.     

Neurochemical changes in rats coexposed to lead and copper

Rats were coexposed to lead (Pb) and Copper (Cu) through drinking water and intraperitoneally, respectively, for a period of 21 days. Neurochemical studies in these rats showed significant reduction in the activity of adenosine triphosphatase, cytochrome-c-oxidase, diaphorase and in the levels of biogenic amines in the rats simultaneously exposed to the two metals compared to either of the metal alone. These neurotoxic effects were not related to the contents of either of the metals in the brain since their accumulation after combined exposure was much less than observed after individual exposure to Pb or Cu.     

Interaction profile for lead, manganese, zinc, and copper

This interaction profile discusses and evaluates the evidence for joint toxic action among lead, manganese, zinc, and copper. The interaction profile recommends how to incorporate concerns about possible interactions or additivity into public health assessments of hazardous waste sites where people might be exposed to mixtures of these chemicals. The profile recommends using endpoint-specific hazard indexes and a hazard quotient to screen for potential health effects. The qualitative weight-of-evidence (WOE) approach is then used to predict the impact of interactions on the endpoint-specific hazard indexes and hazard quotient.     

Succimer treatment and calcium supplementation reduce tissue lead in suckling rats

The effect of combined treatment with meso-2,3-dimercaptosuccinic acid (DMSA) and calcium supplementation in reducing lead absorption and enhancing lead elimination was evaluated in suckling rats under two experimental conditions: during ongoing oral lead exposure (lead acetate, 2 mg Pb kg(-1) day(-1), total dose 16 mg Pb kg(-1)) or after lead exposure (72 h after a 2-day lead exposure, total dose 12 mg Pb kg(-1) s.c.). The artificial feeding method was used for calcium supplementation, with 6% Ca (as CaHPO(4)) suspension in cow's milk to increase the daily calcium intake about three times above control values. Artificial feeding lasted for 7 h a day over eight consecutive days. During this period DMSA was administered on 6 days twice a day (0.5 mmol kg(-1) day(-1) p.o.). At the end of the experiments, Pb, Ca and Zn in the carcass and Pb, Fe and Cu in the liver, kidneys and brain were analysed by atomic absorption spectrometry. Calcium supplementation during lead exposure reduced tissue lead but had no effect when applied after lead exposure, and DMSA administered either during or after lead exposure lowered the tissue lead. Combined treatment during ongoing lead exposure caused a greater reduction in tissue lead than either DMSA or calcium treatment alone. When administered after lead exposure, it had no advantage over DMSA treatment alone but did not impair its efficacy. Combined treatment had no influence on growth and did not seriously disturb essential element status. It is concluded that calcium supplementation could be applied during DMSA therapy, when indicated.     

Effect of zinc on cadmium,copper and zinc contents in cadmium exposed rats

The effect of zinc (Zn) administration on the uptake and retention of cadmium (Cd), copper (Cu) and Zn in liver, kidneys and testes of cadmium exposed rats was investigated. Exposure to Cd caused a significant increase in the uptake of these metals in the 3 organs. The administration of Zn after the Cd exposure had little effect on the level of these metals.     

Marginal zinc deficiency exacerbates bone lead accumulation and high dietary zinc attenuates lead accumulation at the expense of bone density in growing rats.

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.     

Interaction of lead and zinc on the prothrombin activity in rats

In 1955 Saita and coworkers reported a defect in blood coagulation in workers exposed to lead. Very little attention has been given to this observation. In our study in rats receiving 100 ppm lead (acetate) on an adequate semi-purified diet we observed a decreased prothrombin activity. This condition was more severe when dietary zinc was low (5 ppm) than when superoptimal (50 ppm). The correlation between prothrombin time and blood lead was highly significant (r = 0.842, P < 0.001). Although it has been generally accepted that altered prothrombin activity may arise from liver dysfunction, nevertheless serum glutamate-oxaloacetate transaminase (SGOT), a measure of liver function, was normal in all animals. Changes were observed, however, in serum protein pattern due to lead, the A/G ratio was lowered in lead-exposed rats receiving low zinc thus indicating a possible hepatic alteration.     

Succimer treatment during ongoing lead exposure reduces tissue lead in suckling rats.

There is a concern that oral treatment with succimer (meso-2, 3-dimercaptosuccinic acid, DMSA) can promote gastrointestinal lead absorption if not performed in a lead-safe environment. The scope of our investigation was to evaluate the efficacy of oral DMSA treatment during oral lead exposure on tissue lead in suckling rats. Six-day-old Wistar rats of both genders were divided into two groups-untreated (Pb) and treated (Pb + DMSA)-with 10 animals per group. Lead (as acetate) was given orally at a dose of 2 mg kg(-1) body weight day(-1) for eight consecutive days (total dose 16 mg kg(-1), i.e. 0.08 mmol kg(-1)). During this period the treated group received a daily dose of 0.5 mmol DMSA kg(-1) body weight p.o. six times on days 1-3 and 6-8 of the experiment (total dose 3 mmol kg(-1)). Tissue lead was determined by means of atomic absorption spectrometry. The DMSA efficiently reduced the lead concentration in the analysed tissues (carcass, liver, kidneys and brain) by approximately 50% compared with untreated controls. The pups' growth and organ weights were not affected. In conclusion, our results indicate that DMSA is an efficient oral lead chelator in sucklings even if challenged with ongoing lead exposure.     

Influence of zinc supplementation on imipramine effect in a chronic unpredictable stress (CUS) model in rats

Zinc is an endogenous modulator of neuronal activity and may play an important role in the pathogenesis of depression. Recent studies have shown that zinc exhibits antidepressant-like activity in some models of depression in rodents. Our previous studies have shown that the footshock-induced fighting behavior was reduced in the rats subjected to chronic unpredictable stress (CUS). This test is used as the new experimental model of depression. Various antidepressant drugs given repeatedly prevented this kind of behavioral depression. The aim of the present study was to evaluate the effect of prolonged treatment with zinc hydroaspartate and to examine if zinc supplementation could modulate the imipramine effect in CUS model of behavioral depression in rats. The experiments were carried out on male Wistar rats. Chronic stress (persisting for 16 days) was induced by the modified method described by Katz et al. Zinc hydroaspartate at the dose of 30 mg/kg/day or 15 mg/kg/day and imipramine at the dose of 5 mg/kg/day were administered once daily for 14 days. Imipramine was given (ip) 1 h before every stress session and zinc hydroaspartate (ip) l h before the antidepressant. The footshock-induced fighting behavior test was performed 48 h after the last session of the chronic stress. It was demonstrated that in chronically stressed rats the number of fighting attacks was significantly reduced (by about 75%). Zinc hydroaspartate at the dose of 30 mg/kg/day, given alone, prevented the deficit in fighting behavior in chronically stressed rats. Neither imipramine at the dose of 5 mg/kg/day nor zinc hydroaspartate (15 mg/kg/day) administered alone changed the intensity of fighting behavior in chronically stressed rats. However, when imipramine was given at the same dose in the rats pretreated with zinc hydroaspartate (15 mg/kg/day) the deficit of fighting behavior was not observed. The present results indicate that zinc similarly to antidepressants protects the rats against the CUS-induced behavioral depression. Moreover, our findings suggest that zinc supplementation could potentiate the antidepressant effect of imipramine.     

Urinary metallothionein and tissue metal levels of rats injected with cadmium, mercury, lead, copper or zinc

Since Cd exposure results in a dose dependent increase in metallothionein level in urine, the present investigation was conducted to examine whether exposure to other divalent cations would also cause an elevation in urinary metallothionein. Female Sprague-Dawley rats were injected subcutaneously with either saline, 5 mumol/kg/day of CdCl2, HgCl2, Pb(C2H3O2)2, CuSO4 or ZnCl2 for 5 days. Significant increases in hepatic Cu levels in rats treated with not only Cu, but also Zn, Cd, or Hg, and in hepatic Zn levels in rats treated with Zn or Cd were noted. Similarly, renal Cu and Zn levels were elevated significantly in all groups except the Pb-injected group. These increases in tissue metal levels were presumably due to induction of metallothionein. The urinary metallothionein level in control rats on day 0, determined by radioimmunoassay, was 0.85 +/- 0.17 mg/g creatinine. There was no significant change in urinary metallothionein level in rats given up to 5 injections of saline or Pb. Hg-injected rats showed 25-fold increase in urinary metallothionein after 5 injections, whereas Cd-injected rats had 9-fold increase. There were also 2- and 3-fold increases of urinary metallothionein by Cu and Zn treatments for 5 days, respectively. Thus, urinary metallothionein levels were elevated in response to Cd, Hg, Cu and Zn, but not Pb; Hg had the most profound effect at equimolar doses.     

Levels of copper and zinc in depression.

The study was undertaken to estimate plasma copper and zinc in thirty-five depressed patients. Two blood samples were drawn from each patient, one before starting treatment and the second after recovery from depression. The results were compared with the thirty-five normal healthy individuals. The mean plasma copper in controls, depressed patients and in patients after recovery were 106.82, 122.14 and 104.22 micrograms/dl, respectively. The copper levels in patients when depressed were significantly higher as compared to controls and after recovery from depression. The mean plasma zinc levels in controls, depressed patients and after recovery were 115.80, 107.62 and 125.68 micrograms/dl, respectively. No significant difference could be obtained between control and depressed patients. However, the values were significantly higher in recovered patients compared to patients with depression.