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Pyruvate Kinase Activity and δ-Aminolevulinic Acid Dehydratase Activity as Biomarkers of Toxicity in Workers Exposed to Lead

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Abstract

Lead (Pb2+) is a heavy metal that has long been used by humans for a wide range of technological purposes, which is the main reason for its current widespread distribution. Pb2+ is thought to enter erythrocytes through anion exchange and to remain in the cell by binding to thiol groups. Pyruvate kinase (PK) is a thiol-containing enzyme that plays a key role in erythrocyte cellular energy homeostasis. δ-aminolevulinic acid dehydratase (δ-ALAD) is the second enzyme in the heme biosynthetic pathway and plays a role in the pathogenesis of Pb poisoning. Our primary objective was to investigate the effect of Pb2+ on the activity of the thiolenzymes δ-ALAD and PK and on the concentration of glutathione (GSH), a nonenzymatic antioxidant defense, in erythrocytes from Pb-exposed workers. The study sample comprised 22 male Pb workers and 21 normal volunteers (15 men and 6 women). The Pb-exposed workers were employed in manufacturing and recycling of automotive batteries. Basic red-cell parameters were assayed and total white blood cell counts performed. PK and δ-ALAD activity and blood Pb (BPb) concentrations were determined in all subjects. Pb-exposed individuals had significantly greater BPb levels than controls. Both PK and δ-ALAD activity levels were significantly lower in Pb-exposed individuals than in controls. Pb significantly inhibited PK and δ-ALAD activity in a dose-dependent manner. We found that erythrocyte GSH levels were lower in Pb-exposed individuals than normal volunteers. Pb-exposed individuals had lower values than controls for several red cell parameters (hemoglobin, hematocrit, red blood cell count, mean corpuscular volume). These results suggest that Pb inhibits δ-ALAD and PK activity by interacting with their thiol groups. It is therefore possible that Pb disrupts energy homeostasis and may be linked with decreased glucose metabolism because it affects the heme synthesis pathway in erythrocytes, contributing to the cell dysfunction observed in these in Pb-exposed individuals. These results indicate an apparent dose-effect relationship between PK activity and BPb. PK activity in human erythrocytes can be used for biological monitoring of Pb exposure. Study of the mechanisms by which Pb acts may contribute to greater understanding of the symptoms caused by Pb.

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References

  • Agency for Toxic Substances and Disease Registry: Toxicological Profile for Lead (2007) United States Department of Heath and Human Services, Public Health Service, ATSDR

  • Armstrong JS, Jones DP (2002) Glutathione depletion enforces the mitochondrial permeability transition and causes cell death in Bcl-2 overexpressing HL60 cells. FASEB J 16(10):1263–1265

    CAS  Google Scholar 

  • Berlin A, Schaller KH (1974) European standardized method for the determination of delta-aminolevulinic acid dehydratase activity in blood. Z Klin Chem Klin Biochem 12:389–390

    CAS  Google Scholar 

  • Bernard A, Lauwerys R (1987) Metal-induced alterations of delta-aminolevulinic acid dehydratase. Ann N Y Acad Sci 514:41–47

    Article  CAS  Google Scholar 

  • Berry E, Toms NJ (2006) Pyruvate and oxaloacetate limit zinc-induced oxidative HT-22 neuronal cell injury. Neurotoxicology 27(6):1043–1051

    Article  CAS  Google Scholar 

  • Bianchi P, Zanella A (2000) Hematologically important mutations: red cell pyruvate kinase (third update). Blood Cells Mol Dis 26:47–53

    Article  CAS  Google Scholar 

  • Brito JA, McNeill FE, Webber CE, Wells S, Richard N, Carvalho ML et al (2002) Evaluation of a novel structural model to describe the endogenous release of lead from bone. J Environ Monit 4:194–201

    Article  CAS  Google Scholar 

  • Browne RW, Armstrong D (1998) Reduced glutathione and glutathione disulfide. Methods Mol Biol 108:347–352

    CAS  Google Scholar 

  • Calderón-Salinas JV, Hernández-Luna C, Valdez-Anaya B, Maldonado-Vega M, López-Miranda A (1996a) Evolution of lead toxicity in a population of children. Hum Exp Toxicol 15:376–382

    Article  Google Scholar 

  • Calderón-Salinas JV, Valdez-Anaya B, Mazúñiga-Charles Albores-Medina A (1996b) Lead exposure in a population of Mexican children. Hum Exp Toxicol 15(4):305–311

    Article  Google Scholar 

  • Calderón-Salinas JV, Quintanar-Escorza MA, Hernández-Luna CE, González- Martínez MT (1999a) Effect of lead on the calcium transport in human erythrocyte. Hum Exp Toxicol 18(3):146–153

    Article  Google Scholar 

  • Calderón-Salinas JV, Quintanar-Escorza MA, González-Martínez MT, Hernández-Luna CE (1999b) Lead and calcium transport in human erythrocyte. Hum Exp Toxicol 18(5):327–332

    Article  Google Scholar 

  • Courtois E, Marques M, Barrientos A (2003) Lead-induced down-regulation of soluble guanylate cyclase in isolated rat aortic segments mediated by reactive oxygen species and cyclo-oxygenase-2. J Am Soc Nephrol 14:1464–1470

    Article  CAS  Google Scholar 

  • Das UN (2006) Pyruvate is an endogenous anti-inflammatory and anti-oxidant molecule. Int Med J Exp Clin Res 12:79–84

    Google Scholar 

  • Dusinska M, Collins AR (2008) The comet assay in human biomonitoring: gene-environment interactions. Mutagenesis 23:191–205

    Article  CAS  Google Scholar 

  • Ekong EB, Jaar BG, Weaver VM (2006) Lead-related nephrotoxicity: a review of epidemiologic evidence. Kidney Int 70:2074–2084

    CAS  Google Scholar 

  • Ercal N, Gurer-Orhan H, Aykin-Burns N (2011) Toxic metals and oxidative stress. Part 1. Mechanisms involved in metal-induced oxidative damage. Curr Top Med Chem 1:529–539

    Article  Google Scholar 

  • Farant JP, Wigfield DC (1982) Biomonitoring lead exposure with delta-aminolevulinate dehydratase (ALA-D) activity ratios. Int Arch Occup Environ Health 51:15–24

    Article  CAS  Google Scholar 

  • Garza A, Vega R, Soto E (2006) Cellular mechanisms of lead neurotoxicity. Med Sci Monit 12(3):57–65

    Google Scholar 

  • Gilbert HF (1984) Redox control of enzyme activities by thiol/disulfide exchange. Methods Enzymol 107:330–351

    Article  CAS  Google Scholar 

  • Goering PL (1993) Lead–protein interactions as a basis for lead toxicity. Neurotoxicology 14(2–3):45–60

    CAS  Google Scholar 

  • Greenwood NN, Earnshaw A (1997) Chemistry of the elements, 2nd edn. Butterworth-Heinemann, London

    Google Scholar 

  • Gurer H, Ercal N (2000) Can antioxidants be beneficial in the treatment of lead poisoning? Free Radic Biol Med 29:927–945

    Article  CAS  Google Scholar 

  • Gurer H, Neal R, Yang P, Oztezcan S, Ercal N (1999) Captropril as an antioxidant in lead-exposed Fischer 344 rats. Hum Exp Toxicol 18:27–32

    Article  CAS  Google Scholar 

  • Gurer-Ohrnan H, Sabir HU, Ozgünes H (2004) Correlation between clinical indicators of lead poisoning and oxidative stress parameters in controls and lead-exposed workers. Toxicology 195(2–3):147–154

    Article  Google Scholar 

  • Hall ER, Cottam GL (1978) Isoenzymes of pyruvate kinase in vertebrates: their physical, chemical, kinetic and immunological properties. Int J Biochem 9:785–793

    Article  CAS  Google Scholar 

  • Huang M, Krepkiy D, Hu W, Petering DH (2004) Zn-, Cd-, and Pb-transcription factor IIIA: properties, DNA binding, and comparison with TFIIIA finger 3 metal complexes. J Inorg Biochem 98:775–785

    Article  CAS  Google Scholar 

  • Korsrud GO, Meldrum JB (1988) Effect of diet on response in rats to lead acetate given orally or in the drinking water. Biol Trace Elem Res 17:167–173

    Article  CAS  Google Scholar 

  • Lauwerys R, Buchet JP, Roels HA, Materne DO (1974) Relationship between urinary delta-aminolevulinic acid excretion and the inhibition or read cell delta-aminolevulinate dehydratase by lead. Clin Toxicol 7:383–388

    Article  CAS  Google Scholar 

  • Lawton LJ, Donaldson WE (1991) Lead-induced tissue fatty acid alterations and lipid peroxidation. Biol Trace Elem Res 28:83–97

    Article  CAS  Google Scholar 

  • Leech NL, Barrett KC, Morgan GA (2005) SPSS for intermediate statistics. Use and interpretation, 2nd edn. Lawrence Erlbaum Associates, London

    Google Scholar 

  • Leong SF, Lai JCK, Lim L, Clark JB (1981) Energy-metabolising enzymes in brain regions of adult and aging rats. J Neurochem 37:1548–1556

    Article  CAS  Google Scholar 

  • Lepper TW, Evandro Oliveira E, Koch GDW, Berlese DB, Feksa LR (2010) Lead inhibits in vitro creatine kinase and pyruvate kinase activity in brain cortex of rats. Toxicol in Vitro 24:1045–1051

    Article  CAS  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  Google Scholar 

  • Meister A (1994) Glutathione, ascorbate, and cellular protection. Cancer Res 54:1969–1975

    Google Scholar 

  • Menke A, Muntner P, Batuman P, Silbergeld EK, Guallar E (2006) Blood lead below 0.48 μmol/L (100 μg L−1) and mortality among US adults. Circulation 114:1388–1394

    Article  CAS  Google Scholar 

  • Meredith PA, Moore MR, Campbell BC, Thompson GG, Goldberg A (1978) Delta-aminolevulinic acid metabolism in normal and lead-exposed humans. Toxicology 9:1–9

    Article  CAS  Google Scholar 

  • Muntener P, He J, Vupputuri S, Coresh J, Batuman V (2003) Blood lead and chronic kidney disease in the general United States population: results from NHANES III. Kidney Int 63:1044–1050

    Article  Google Scholar 

  • Nash D, Magder L, Lustberg M, Sherwin RW, Rubin RJ, Kaufmann RB, Silbergeld EK (2003) Blood lead, blood pressure, and hypertension in perimenopausal and postmenopausal women. JAMA 289(12):1523–1532

    Google Scholar 

  • Navarro-Moreno LG, Quintanar-Escorza MA, González S, Mondragón R, Cerbón-Solorzáno J, Valdés J et al (2009) Effects of lead intoxication on intercellular junctions and biochemical alterations of the renal proximal tubule cells. Toxicol In Vitro 23:1298–1304

    Article  CAS  Google Scholar 

  • Noguchi T, Yamada K, Inoue H, Matsuda T, Tanaka T (1987) The L- and R-type isozymes of rat pyruvate kinase are produced from a single gene by use of different promoters. J Biol Chem 262:14366–14371

    CAS  Google Scholar 

  • Nunes-Tavares N, Valverde RHF, Araújo GMN, Hassón-Voloch A (2005) Toxicity induced by Hg2+ on choline acetyltransferase activity from E. electricus (L.) electrocytes: the protective effect of 2,3 dimercapto-propanol (BAL). Med Sci Monit 11(4):100–105

    Google Scholar 

  • Occupational Safety and Health Administration (2008) Employee standard summary—1910.1025 App B [displayed 6 January 2008]. Available at: http://www.osha.gov/Publications/OSHA3348-metal-scrap-recycling.pdf

  • Olewińska E, Kasperczyk A, Kapka L, Kozłowska A, Pawlas N, Dobrakowski M et al (2010) Level of DNA damage in lead-exposed workers. Ann Agric Environ Med 17:231–236

    Google Scholar 

  • Onalaja AO, Claudio L (2000) Genetic susceptibility to lead poisoning. Environ Health Perspect 108:23–28

    CAS  Google Scholar 

  • Quintanar-Escorza MA, González-Martínez MT, Navarro L, Maldonado M, Arévalo B, Calderón-Salinas JV (2007) Intracellular free calcium concentration and calcium transport in human erythrocyte of lead-exposed workers. Toxicol Appl Pharmacol 220:1–8

    Article  CAS  Google Scholar 

  • Reed DJ (1990) Glutathione: toxicological implications. Annu Rev Pharmacol Toxicol 30:603–631

    Article  CAS  Google Scholar 

  • Rendón-Ramírez A, Cerbón-Solórzano J, Maldonado-Vega M, Quintanar-Escorza MA, Calderón-Salinas JV (2007) Vitamin-E reduces the oxidative damage on delta-aminolevulinic dehydratase induced by lead intoxication in rat erythrocytes. Toxicol in Vitro 21(6):1121–1126

    Article  Google Scholar 

  • Ribarov SR, Benov LC (1981) Relationship between the hemolytic action of heavy metals and lipid peroxidation. Biochim Biophys Acta 640:721–726

    Article  CAS  Google Scholar 

  • Sakai T (2000) Biomarkers of lead exposure. Ind Health 38:127–142

    Article  CAS  Google Scholar 

  • Selander S, Cramer K (1970) Interrelationship between lead in blood, lead in urine and ALA in urine during lead worse. Br J Ind Med 27:28–39

    CAS  Google Scholar 

  • Sithisarankul P, Schwartz BS, Lee BK, Strickland PT (1998) Urinary 5-aminolevuinic acid (ALA) adjusted by creatinine a surrogate for plasma ALA. J Occup Environ Med 40:901–908

    Article  CAS  Google Scholar 

  • Todd AC, Wetmur JG, Moline JM, Godbold JH, Levin SM, Landrigan PJ (1996) Unraveling the chronic toxicity of lead: an essential priority for environmental health. Environ Health Perspect 104:141–146

    CAS  Google Scholar 

  • Valle BL, Ulmer DD (1972) Biochemical effects of mercury, cadmium and lead. Annu Rev Biochem 41:91–128

    Article  Google Scholar 

  • van Bemmel DM, Boffetta P, Liao LM, Berndt SI, Menashe I, Yeager M et al (2011) Comprehensive analysis of 5-aminolevulinic acid dehydrogenase (ALAD) variants and renal cell carcinoma risk among individuals exposed to lead. PLoS One 6(7):1–7

    Google Scholar 

  • Warren MJ, Cooper JB, Wood SP, Shoolingin JPM (1998) Lead poisoning, haem synthesis and 5-aminolaevulinic acid dehydratase. Trends Biochem Sci 23:217–221

    Article  CAS  Google Scholar 

  • World Health Organization (1995) International Programme on Chemical Safety Environmental Health criteria 165: Inorganic lead. WHO, Geneva

    Google Scholar 

  • Wu G, Fang YZ, Yang S, Lupton JR, Turner ND (2004) Glutathione metabolism and its implications for health. J Nutr 134:489–492

    CAS  Google Scholar 

Download references

Acknowledgments

This research was partly supported by grants from Feevale University, CNPq, FAPERGS, and PROPESQ/UFRGS, Brazil.

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Authors and Affiliations

  1. Instituto de Ciências da Saúde, Grupo de Pesquisa em Bioanálise, Universidade Feevale, RS 239, 2755, Novo Hamburgo, RS, CEP 93352-000, Brazil

    Luciane Rosa Feksa, Evandro Oliveira, Thereza Trombini, Mateus Luchese, Saiuri Bisi, Rafael Linden, Daiane Bolzan Berlese & Larissa Machado Lacerda

  2. Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Luciane Rosa Feksa, Denise Bertin Rojas, Rodrigo Binkowski Andrade, Patricia Fernanda Schuck, Moacir Wajner & Clovis Milton Duval Wannmacher

  3. Laboratório de Erros Inatos do Metabolismo, Universidade do Extremo Sul Catarinense, Criciúma, SC, Brazil

    Patricia Fernanda Schuck

  4. Departamento de Tecnologia e Ciência dos Alimentos, CCR, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil

    Tatiana Emanuelli

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  1. Luciane Rosa Feksa
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Correspondence to Luciane Rosa Feksa.

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Feksa, L.R., Oliveira, E., Trombini, T. et al. Pyruvate Kinase Activity and δ-Aminolevulinic Acid Dehydratase Activity as Biomarkers of Toxicity in Workers Exposed to Lead. Arch Environ Contam Toxicol 63, 453–460 (2012). https://doi.org/10.1007/s00244-012-9786-z

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