Inhibition Of Delta-aminolevulinic Acid Dehydratase Research Articles

Effect ofCentella asiatica on arsenic induced oxidative stress and metal distribution in rats

Concomitant oral supplementation of Centella asiatica (100, 200 or 300 mg kg(-1), orally once daily) during arsenic exposure (20 ppm in drinking water for 4 weeks) was investigated in rats for its protective value. The animals exposed to arsenic (III) showed a significant inhibition of delta-aminolevulinic acid dehydratase (ALAD) activity, a marginal decrease in glutathione (GSH) and an increase in zinc protoporphyrin (ZPP) level in blood. Hepatic and renal glutathione (GSH) decreased, while oxidized glutathione (GSSG) and thiobarbituric acid reactive substance (TBARS) levels increased significantly in the liver, kidney and brain. The activities of brain superoxide dismutase (SOD) and catalase decreased marginally on arsenic exposure. Concomitant administration of Centella asiatica showed a significant protective action on inhibited blood ALAD activity and restored the blood GSH level, whereas most of the other blood biochemical parameters remained unchanged on Centella asiatica supplementation. Interestingly, most of the hepatic biochemical variables indicative of oxidative stress showed protection. There was, however, a significant protection observed in the altered kidney GSSG level and hepatic and brain TBARS. Only a marginal beneficial effect of Centella asiatica on blood and liver arsenic concentration was noted, particularly at the highest dose studies (300 mg kg(-1)). No effect of Centella asiatica on most of the altered renal biochemical parameters was noted. The results thus lead to the conclusion that simultaneous supplementation of Centella asiatica significantly protects against arsenic-induced oxidative stress but does not influence the arsenic concentration in these organs. It can thus be suggested that co-administration of Centella asiatica protects animals from arsenic-induced oxidative stress but exhibits no chelating property. Further studies are recommended for determining the effect of co-administration of Centella asiatica during chelation therapy with a thiol chelator.

Protective value of Aloe vera against some toxic effects of arsenic in rats

Concomitant oral supplementation of Aloe vera, (1, 2 or 5% w[sol ]v in drinking water) during arsenic exposure (0.2 mg[sol ]kg, intraperitoneally, once daily for 3 weeks) was investigated in rats for its protective value. Animals exposed to arsenic (III) showed a significant inhibition of delta-aminolevulinic acid dehydratase (ALAD) activity, a marginal decrease in glutathione (GSH) and an increase in zinc protoporphyrin (ZPP) level in blood. White blood corpuscles (WBC) level decreased while most of the other clinical blood parameters like red blood cells count, haemoglobin, MCV, MCH, MCHC ratio and platelet number, etc. remained unaltered on arsenic exposure. Hepatic reduced GSH, oxidized glutathione (GSSG) level remained unaltered, thiobarbituric acid reactive substance (TBARS) level increased significantly while the activity of alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and catalase decreased on arsenic exposure. Renal GSH contents decreased while superoxide dismutase (SOD) activity decreased significantly on arsenic exposure. Concomitant administration of Aloe vera had remarkable protective action on inhibited blood ALAD activity and restored blood GSH level while most of the other blood biochemical parameters remained unchanged on Aloe vera supplementation. Interestingly, most of hepatic biochemical variables indicative of oxidative stress showed protection; no effect of Aloe vera on blood and liver arsenic concentration was noted. Also, no effect of Aloe vera on most of the altered renal biochemical parameters were noticed. The results thus lead us to conclude that simultaneous supplementation of Aloe vera protects against arsenic induced oxidative stress but does not influence the arsenic concentration in these organs.

Preventive effect of Coriandrum sativum (Chinese parsley) on localized lead deposition in ICR mice

The preventive effect of Coriandrum sativum, Fam. umbelliferae (Chinese parsley) on lead deposition was investigated in male ICR mice given lead (1000 ppm) as lead acetate trihydrate in drinking water for 32 days. Administration of Chinese parsley to mice by gastric intubation was performed for 25 days from day 7 after the start of lead exposure up to the end of the experiment. The mice were then sacrificed for comparison of lead distribution. The lead reached its highest concentration in the femur but localized lead deposition in the femur was significantly decreased by meso-2,3-dimercaptosuccinic acid (DMSA), a chelating agent used as a positive control to validate this experimental model. Administration of Chinese parsley also significantly decreased lead deposition in the femur and severe lead-induced injury in the kidneys. In addition, urinary excretion of delta-aminolevulinic acid (ALA) which is known to increase with lead intake was significantly decreased after administration of Chinese parsley. The MeOH extract of Chinese parsley also reduced lead-induced inhibition of delta-aminolevulinic acid dehydratase (ALAD) activity in vitro. These results suggest that Chinese parsley has suppressive activity on lead deposition, probably resulting from the chelation of lead by some substances contained in Chinese parsley.

Biomarkers of lead exposure.

Biomarkers of exposure, effect, and susceptibility are reviewed in relation to lead exposure. Of the biomarkers of lead exposure, blood lead (Pb-B), mainly red cell lead, is a representative of soft tissue lead, and most widely used as measures of body burden and absorbed (internal) doses of lead. Urine lead (Pb-U) as well as plasma lead (Pb-P) increases exponentially with increasing Pb-B under a steady-state situation and is a reflection of recent exposure. The amount of lead in plasma and urine (MPb-P and MPb-U) after administration of a chelating agent (e.g. CaEDTA) can be useful for biomarkers of internal exposure of lead, reflecting the mobilizable pool of lead which consists of mainly blood and soft tissue lead with only a small fraction derived from bones. The critical effects in bone marrow arise mainly from the interaction of lead with some enzymatic process responsible for heme synthesis. The effects can be used for the biomarkers of effects. They are the inhibition of delta-aminolevulinic acid dehydratase (ALAD) and the variation in some metabolite concentrations (e.g. delta-aminolevulinic acid in urine (ALA-U), blood (ALA-B) or plasma (ALA-P), coproporphyrin in urine (CP), zinc protoporphyrin (ZP) in blood). The activities of pyrimidine nucleotidase (P5'N) and nicotinamide adenine dinucleotide synthetase (NADS) in blood are also decreased in lead exposure, and nucleotide contents in blood is altered in lead exposure. These effects of lead on human can be also useful biomarkers of effect. The differences in levels of heme precursors between two types of ALAD genotypes might be attributable to those in the affinity of different ALAD isozymes to lead. ALAD1 homozygotes have higher levels of ZP and ALA in comparison with ALAD2 carriers at the high lead exposure, suggesting that ALAD1 homozygotes might be more susceptible for disturbance in heme biosynthesis by lead than ALAD2 carriers.

A comparison of the effects of inorganic and alkyllead compounds on human erythrocytic δ‐aminolevulinic acid dehydratase (ALAD) activity in vitro

A study comprising two experiments comparing the effects of the alkyllead compounds triethyllead acetate and tetraethyllead as well as an inorganic lead compound (Pb(NO3)2) on human erythrocytic delta-aminolevulinic acid dehydratase activity was undertaken. In Experiment 1, four human blood samples were pre-incubated for 0 min or 60 min with increasing concentrations (0-1000 microM) of triethyllead acetate, tetraethyllead and Pb(NO3)2, and the delta-aminolevulinic acid dehydratase activity was determined. There was no discernible inhibitory effect of tetraethyllead at any concentration, with or without pre-incubation. Pb(NO3)2 showed a distinct effect at an addition concentration of 1 microM, with a substantial inhibition only following pre-incubation. Triethyllead acetate showed no difference in effect with pre-incubation. In the second experiment, the activating effects of zinc chloride (60 microM) and dithiothreitol (20 mM) on delta-aminolevulinic acid dehydratase activity were examined on four human blood samples in the presence of varying concentrations of Pb(NO3)2 (0-50 microM) and triethyllead acetate (0-1000 microM). The addition of zinc chloride alone showed little effect on enzyme activity, while dithiothreitol alone caused a substantial activation of the enzyme at all lead concentrations. The addition of both zinc chloride and dithiothreitol caused an even greater activation of the enzyme. The data indicate that Pb(NO3)2 is a much more potent inhibitor of delta-aminolevulinic acid dehydratase activity than triethyllead acetate (Ki values of 0.77 microM versus 130.37 microM. respectively). The use of delta-aminolevulinic acid dehydratase activity as an indicator of exposure may be questionable in cases of alkyllead exposure. Possible causes for differences in the inhibition of delta-aminolevulinic acid dehydratase by these compounds is also discussed.

Interaction of Zinc, Methionine or Their Combination with Lead at Gastrointestinal or Post‐Absorptive Level in Rats

The ability of zinc, methionine or their combination given by gavage to prevent or treat experimental oral lead intoxication in rats was investigated. Simultaneous oral supplementation with zinc plus methionine was found to be most effective in reducing lead induced inhibition of delta-aminolevulinic acid dehydratase (ALAD) activity in blood, elevation of urinary delta-aminolevulinic acid (ALA) excretion and in enhancing the hepatic glutathione (GSH) contents. The combination was also most effective in reducing the accumulation of lead in blood, liver and kidney compared to zinc or methionine alone. Prevention was more effective than treatment after lead exposure which may be caused by a decrease in the absorption of lead in the gastrointestinal tract in the presence of zinc and/or methionine.

Inhibition of liver, kidney, and erythrocyte δ-aminolevulinic acid dehydratase (porphobilinogen synthase) by gallium in the rat

Selective inhibition of enzymes in the heme biosynthesis pathway with concomitant urinary excretion of heme precursors serve as potentially important biological markers of chemical exposure and cell injury. Intratracheal administration of gallium arsenide particulate suspensions has been shown to result in inhibition of delta-aminolevulinic acid dehydratase (ALAD) in several tissues and increased excretion of the heme precursor aminolevulinic acid (ALA). This study was undertaken to evaluate in vivo the role of gallium alone in ALAD inhibition and increased urinary excretion of ALA. Male CD rats received a single ip injection of Ga2(SO4)3 at doses of 12.5, 25, 50, 100, and 200 mg Ga/kg. A dose-dependent inhibition of ALAD was observed 24 hr later in liver, kidney, and erythrocytes. After injection of 25 mg Ga/kg, maximal inhibition (42 to 49% of control) of ALAD occurred between 6 and 24 hr in liver and kidney with full recovery of activity at 96 hr. In erythrocytes, maximal inhibition (48% of control) occurred between 24 and 48 hr with recovery of activity at 96 hr. Mild to moderate renal proximal tubular necrosis in the pars recta was observed 24 hr after administration of 100 and 200 mg/kg, but no histopathologic changes were evident at lower doses. No consistent changes in urinary excretion of ALA were observed. Lineweaver-Burk analyses of renal and hepatic ALAD activities in the absence and presence of gallium indicated that the inhibition of ALAD by this element is noncompetitive (same Km, decreased Vmax). Gallium was shown to possess an inhibition constant (Ki) of approximately 3 microns for ALAD, similar to the Ki obtained for lead in other studies. Incubation of ALAD in vitro with gallium and lead, an active thiol group inhibitor, resulted in a greater inhibition of the enzyme. Further in vitro studies demonstrated the attenuation of gallium inhibition of hepatic and renal ALAD by zinc, suggesting that the mechanism of gallium action may involve competition for or displacement of zinc from the sulfhydryl group of the enzyme active site. Since ALAD inhibition occurred at doses at which no histopathologic changes were evident, the determination of ALAD activity in various tissues, including blood, may be of potential value as a biomarker of exposure/toxicity to metals such as gallium. The effect of chemical form and route of exposure of gallium and effects of other Group III metals on inhibition of ALAD and excretion of ALA is discussed.(ABSTRACT TRUNCATED AT 250 WORDS)

Microbial production of vitamin B12 antimetabolites. IV. Isolation and identification of 4-keto-5-amino-6-hydroxyhexanoic acid.

4-Keto-5-amino-6-hydroxyhexanoic acid was isolated from Bacillus cereus 102804 fermentations and found to inhibit the growth of Gram-positive and Gram-negative bacteria, when grown in a chemically defined medium. The mechanism appeared to be the inhibition of delta-aminolevulinic acid dehydratase. The Ki value of 4-keto-5-amino-6-hydroxyhexanoic acid in an enzyme preparation of Propionibacterium shermanii was 0.72 microM. Similar test conditions with 4-keto-5-aminohexanoic acid resulted in Ki of 12.1 microM. In both cases competitive inhibition was found. The structure of 4-keto-5-amino-6-hydroxyhexanoic acid was determined.

Antagonistic effects of zinc and aluminum on lead inhibition of delta-aminolevulinic acid dehydratase.

In vitro and in vivo studies regarding the influence of metals on delta-aminolevulinic acid dehydratase activity in erythrocytes indicate that a lead concentration of 4 micrometer/l completely inhibits the enzyme. Zinc activates the enzyme both in vitro and in vivo at concentrations greater than 76 micrometer/l. Aluminum has an inhibitory effect at all concentrations in vitro whereas it activates the enzyme in vivo. Zinc and aluminum together activate the enzyme in vivo. The in vitro activation of zinc is inhibited by increasing concentrations of aluminum. Aluminum and lead together depress the enzyme activity in an additive way that can be reactivated by the addition of zinc.

Inhibition of delta-aminolevulinic acid dehydratase in human red blood cells by lead and activation by zinc or cysteine.

Inhibition of blood delta-aminolevulinic acid dehydratase(ALA-D) activity by lead was studied in vivo and in vitro. In vivo, a negative linear correlation (r = -0.85) was found between the logarithmic values of ALA-D activity and blood lead levels. In vitro the inhibitory effect of lead on blood ALA-D activity increased both with contact time and contact temperature of lead with blood before ALA-D assay. Maximum enzyme inhibition occurred after 14 h of contact at 25 degrees C. Inhibition of ALA-D activity by lead, in vivo as well as in vitro, is suppressed by the addition of zinc or cysteine. The logarithmic values of the activity ratios increase linearly with blood lead concentrations. The increase in ALA-D activity brought about by the addition of zinc or cysteine can be used to identify cases of low enzyme activity with no lead intoxication involved. The same technique can also detect cases in which ALA-D inhibition may be concealed by a presumably high initial enzyme activity as observed in some patients.

Lead intoxication among demolition workers: The effect of lead on the hepatic cytochrome P‐450 system in humans

Overt clinical disease from undue lead exposure has become a relatively rare phenomenon in adult populations. However, exposure situations that may result in subclinical disease are not uncommon in various occupational settings. Five demolition workers, dismantling an old iron structure covered with lead-content paint, were studied. The use of cutting torches resulted in lead fumes, with significant exposure, albeit without gross 'lead poisoning." All five workers showed biochemical manifestations of chronic lead intoxication-that is, elevated blood lead level, inhibition of delta-aminolevulinic acid dehydratase (ALA-D), and elevated erythrocytic protoporphyrin concentration (PROTO). The effect of lead on the biosynthesis of heme was assessed by investigating the functional capacity of the cytochrome P-45O system of the liver, through drug metabolism studies. The plasma elimination rates (half-lives) of antipyrine and phenylbutazone-drugs primarily metabolized by the hemeprotein P-450 dependent hepatic microsomal enzyme system-were measured before and after chelation therapy. Prior to chelation therapy all half-lives were within the normal range. A slight decrease in the half-life of antipyrine was found after treatment. These studies show that chronic exposure to lead has only a minimal effect on hepatic cytochrome p-450 dependent enzymatic activities in adult males.