Metabolic Brain Disease

, Volume 31, Issue 5, pp 1123–1132 | Cite as

Protective effects of ascorbic acid and garlic extract against lead-induced apoptosis in developing rat hippocampus

  • Ali-Reza Ebrahimzadeh-Bideskan
  • Javad Hami
  • Fatemeh Alipour
  • Hossein Haghir
  • Ali-Reza Fazel
  • Akram Sadeghi
Original Article


Lead exposure has negative effects on developing nervous system and induces apoptosis in newly generated neurons. Natural antioxidants (i.e. Ascorbic acid and Garlic) might protect against lead-induced neuronal cell damage. The aim of the present study was to investigate the protective effects of Ascorbic acid and Garlic administration during pregnancy and lactation on lead-induced apoptosis in rat developing hippocampus. Timed pregnant Wistar rats were administrated with Lead (1500 ppm) via drinking water (Pb group) or lead plus Ascorbic acid (Pb + AA Group, 500 mg/kg, IP), or lead plus Garlic Extract (Pb + G Group, 1 ml garlic juice/100 g BW, via Gavage) from early gestation (GD 0) until postnatal day 50 (PN 50). At the end of experiments, the pups’ brains were carefully dissected. To identify neuronal death, the brain sections were stained with TUNEL assay. Mean of blood and brain lead levels increased significantly in Pb group comparing to other studied groups (P < 0.01). There was significant reduction in blood and brain lead level in Pb + AA and Pb + G groups when compared to those of Pb group (P < 0.01). The mean number of TUNEL positive cells in the CA1, CA3, and DG was significantly lower in the groups treated by either Ascorbic acid or Garlic (P < 0.05). Administration of Ascorbic acid and Garlic during pregnancy and lactation protect against lead-induced neuronal cell apoptosis in the hippocampus of rat pups partially via the reduction of Pb concentration in the blood and in the brain.


Hippocampus Lead Ascorbic acid Garlic 



This paper is based on the results of Mrs. Akram Sadeghi M.Sc. thesis which financially was supported by vice chancellor for research, Mashhad University of Medical Sciences (MUMS) grant (no. 89230). The authors gratefully thanks to Mrs. Motejadded for her technical assistance.


  1. Abusaad I, MacKay D, Zhao J, Stanford P, Collier DA, Everall IP (1999) Stereological estimation of the total number of neurons in the murine hippocampus using the optical disector. J Comp Neurol 408:560–566CrossRefPubMedGoogle Scholar
  2. Agus DB, Gambhir SS, Pardridge WM, Spielholz C, Baselga J, Vera JC, Golde DW (1997) Vitamin C crosses the blood-brain barrier in the oxidized form through the glucose transporters. J Clin Invest 100:2842–2848Google Scholar
  3. Ahamed M, Siddiqui MK (2007) Low level lead exposure and oxidative stress: current opinions. Clin Chim Acta 383:57–64. doi: 10.1016/j.cca.2007.04.024 CrossRefPubMedGoogle Scholar
  4. Alfano DP, Petit TL (1981) Behavioral effects of postnatal lead exposure: possible relationship to hippocampal dysfunction. Behav Neural Biol 32:319–333CrossRefPubMedGoogle Scholar
  5. Altmann L, Weinsberg F, Sveinsson K, Lilienthal H, Wiegand H, Winneke G (1993) Impairment of long-term potentiation and learning following chronic lead exposure Toxicol Lett 66:105–112PubMedGoogle Scholar
  6. Antonio-Garcia MT, Massó-Gonzalez EL (2008) Toxic effects of perinatal lead exposure on the brain of rats: involvement of oxidative stress and the beneficial role of antioxidants. Food Chem Toxicol 46:2089–2095. doi: 10.1016/j.fct.2008.01.053
  7. Babu MS, Gopal NV, Reddy KP (2007) Post natal antioxidant enzyme activity of rat brain regions during developmental lead exposure. J Environ Biol 28:21–27PubMedGoogle Scholar
  8. Bellinger DC (2007) Lead neurotoxicity in children: decomposing the variability in dose-effect relationships. Am J Ind Med 50:720–728. doi: 10.1002/ajim.20438 CrossRefPubMedGoogle Scholar
  9. Bellinger D, Sloman J, Leviton A, Rabinowitz M, Needleman HL, Waternaux C (1991) Low-level lead exposure and children's cognitive function in the preschool years. Pediatrics 87:219–227PubMedGoogle Scholar
  10. Borek C (2001) Antioxidant health effects of aged garlic extract. J Nutr 131:1010S–1015SPubMedGoogle Scholar
  11. Burdette LJ, Goldstein R (1986) Long-term behavioral and electrophysiological changes associated with lead exposure at different stages of brain development in the rat. Dev Brain Res 29:101–110CrossRefGoogle Scholar
  12. Canfield RL, Henderson CR Jr, Cory-Slechta DA, Cox C, Jusko TA, Lanphear BP (2003) Intellectual impairment in children with blood lead concentrations below 10 microg per deciliter. N Engl J Med 348:1517–1526. doi: 10.1056/NEJMoa022848348/16/1517 CrossRefPubMedPubMedCentralGoogle Scholar
  13. Chang BJ et al. (2012) Ascorbic acid ameliorates oxidative damage induced by maternal low-level lead exposure in the hippocampus of rat pups during gestation and lactation. Food Chem Toxicol 50:104–108. doi: 10.1016/j.fct.2011.09.043 CrossRefPubMedGoogle Scholar
  14. Chen A, Dietrich KN, Ware JH, Radcliffe J, Rogan WJ (2005) IQ and blood lead from 2 to 7 years of age: are the effects in older children the residual of high blood lead concentrations in 2-year-olds? Environ Health Perspect 113:597–601CrossRefPubMedPubMedCentralGoogle Scholar
  15. Chen YH, Xu DX, Zhao L, Wang H, Wang JP, Wei W (2006) Ascorbic acid protects against lipopolysaccharide-induced intra-uterine fetal death and intra-uterine growth retardation in mice. Toxicology 217:39–45. doi: 10.1016/j.tox.2005.08.010 CrossRefPubMedGoogle Scholar
  16. Chung LY (2006) The antioxidant properties of garlic compounds: allyl cysteine, alliin, allicin, and allyl disulfide. J Med Food 9:205–213. doi: 10.1089/jmf.2006.9.205 CrossRefPubMedGoogle Scholar
  17. Colin-Gonzalez AL, Santana RA, Silva-Islas CA, Chanez-Cardenas ME, Santamaria A, Maldonado PD (2012) The antioxidant mechanisms underlying the aged garlic extract- and S-allylcysteine-induced protection. Oxid Med Cell Longev 2012:907162. doi: 10.1155/2012/907162 CrossRefPubMedPubMedCentralGoogle Scholar
  18. Coscia JM, Ris MD, Succop PA, Dietrich KN (2003) Cognitive development of lead exposed children from ages 6 to 15 years: an application of growth curve analysis. Child Neuropsychol 9:10–21. doi: 10.1076/chin. CrossRefPubMedGoogle Scholar
  19. Crowe A, Morgan EH (1996) The effects of iron loading and iron deficiency on the tissue uptake of 64Cu during development in the rat. Biochim Biophys Acta 1291:53–59CrossRefPubMedGoogle Scholar
  20. Davis LE, Shen JK, Cai Y (1990) Antifungal activity in human cerebrospinal fluid and plasma after intravenous administration of Allium sativum. Antimicrob Agents Chemother 34:651–653Google Scholar
  21. Dietrich KN, Succop PA, Berger OG, Hammond PB, Bornschein RL (1991) Lead exposure and the cognitive development of urban preschool children: the Cincinnati Lead Study cohort at age 4 years. Neurotoxicol Teratol 13:203–211CrossRefPubMedGoogle Scholar
  22. Dietrich KN, Berger OG, Succop PA, Hammond PB, Bornschein RL (1993) The developmental consequences of low to moderate prenatal and postnatal lead exposure: intellectual attainment in the Cincinnati Lead Study Cohort following school entry. Neurotoxicol Teratol 15:37–44CrossRefPubMedGoogle Scholar
  23. Dietrich KN et al. (2004) Effect of chelation therapy on the neuropsychological and behavioral development of lead-exposed children after school entry. Pediatrics 114:19–26CrossRefPubMedGoogle Scholar
  24. Drew CA, Spence I, Johnston GA (1990) Effect of chronic exposure to lead on GABA binding in developing rat brain. Neurochem Int 17:43–51CrossRefPubMedGoogle Scholar
  25. Dribben WH, Creeley CE, Farber N (2011) Low-level lead exposure triggers neuronal apoptosis in the developing mouse brain. Neurotoxicol Teratol 33:473–480. doi: 10.1016/ CrossRefPubMedPubMedCentralGoogle Scholar
  26. Eichenbaum H (2004) Hippocampus: cognitive processes and neural representations that underlie declarative memory. Neuron 44:109–120. doi: 10.1016/j.neuron.2004.08.028S089662730400529X CrossRefPubMedGoogle Scholar
  27. Ennever FK (1994) Metals. In: Hayes AW (ed) Principles and method of toxicology. Raven Press, New York, pp. 417–446Google Scholar
  28. Finkelstein Y, Markowitz ME, Rosen JF (1998) Low-level lead-induced neurotoxicity in children: an update on central nervous system effects. Brain Res Brain Res Rev 27:168–176CrossRefPubMedGoogle Scholar
  29. Gautam P, Flora SJ (2010) Oral supplementation of gossypin during lead exposure protects alteration in heme synthesis pathway and brain oxidative stress in rats. Nutrition 26:563–570. doi: 10.1016/j.nut.2009.06.008 CrossRefPubMedGoogle Scholar
  30. Gavrieli Y, Sherman Y, Ben-Sasson SA (1992) Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119:493–501CrossRefPubMedGoogle Scholar
  31. Gilbert ME, Lasley SM (2002) Long-term consequences of developmental exposure to lead or polychlorinated biphenyls: Synaptic transmission and plasticity in the rodent CNS. Environ Toxicol Pharmacol 12:105–117CrossRefPubMedGoogle Scholar
  32. Han JM et al. (2007) Protective effects of ascorbic acid against lead-induced apoptotic neurodegeneration in the developing rat hippocampus in vivo. Brain Res 1185:68–74. doi: 10.1016/j.brainres.2007.09.044 CrossRefPubMedGoogle Scholar
  33. Iciek M, Kwiecien I, Wlodek L (2009) Biological properties of garlic and garlic-derived organosulfur compounds. Environ Mol Mutagen 50:247–265. doi: 10.1002/em.20474 CrossRefPubMedGoogle Scholar
  34. Imai J, Ide N, Nagae S, Moriguchi T, Matsuura H, Itakura Y (1994) Antioxidant and radical scavenging effects of aged garlic extract and its constituents. Planta Med 60:417–420. doi: 10.1055/s-2006-959522 CrossRefPubMedGoogle Scholar
  35. Jedrychowski W et al. (2009) Very low prenatal exposure to lead and mental development of children in infancy and early childhood: Krakow prospective cohort study. Neuroepidemiology 32:270–278. doi: 10.1159/000203075 CrossRefPubMedPubMedCentralGoogle Scholar
  36. Jiao J, Lu G, Liu X, Zhu H, Zhang Y (2011) Reduction of blood lead levels in lead-exposed mice by dietary supplements and natural antioxidants. J Sci Food Agric 91:485–491. doi: 10.1002/jsfa.4210 CrossRefPubMedGoogle Scholar
  37. Kiran Kumar B, Prabhakara Rao Y, Noble T, Weddington K, McDowell VP, Rajanna S, Bettaiya R (2009) Lead-induced alteration of apoptotic proteins in different regions of adult rat brain. Toxicology letters 184:56–60. doi: 10.1016/j.toxlet.2008.10.023 CrossRefPubMedGoogle Scholar
  38. Kumaran D, Maguire EA (2005) The human hippocampus: cognitive maps or relational memory? J Neurosci 25:7254–7259. doi: 10.1523/JNEUROSCI.1103-05.2005 CrossRefPubMedGoogle Scholar
  39. Leelarungrayub N, Rattanapanone V, Chanarat N, Gebicki JM (2006) Quantitative evaluation of the antioxidant properties of garlic and shallot preparations. Nutrition 22:266–274. doi: 10.1016/j.nut.2005.05.010 CrossRefPubMedGoogle Scholar
  40. Lewin G, Popov I (1994) Antioxidant effects of aqueous garlic extract. 2nd communication: Inhibition of the Cu(2+)-initiated oxidation of low density lipoproteins. Arzneimittelforschung 44:604–607PubMedGoogle Scholar
  41. Lidsky TI, Schneider JS (2003) Lead neurotoxicity in children: basic mechanisms and clinical correlates. Brain 126:5–19CrossRefPubMedGoogle Scholar
  42. Lu J, Zhang L, Dai Y, Liu G (2002) [Effects of low-level lead exposure on neurobehaviour development in 1–3 year-old children and the intervention guideline] Wei Sheng Yan Jiu 31:4–6Google Scholar
  43. Lu X et al. (2013) Prenatal and lactational lead exposure enhanced oxidative stress and altered apoptosis status in offspring rats' hippocampus. Biol Trace Elem Res 151:75–84. doi: 10.1007/s12011-012-9531-5 CrossRefPubMedGoogle Scholar
  44. Machartova V, Racek J, Kohout J, Senft V, Trefil L (2000) Effect of antioxidant therapy on indicators of free radical activity in workers at risk of lead exposure. Vnitr Lek 46:444–446PubMedGoogle Scholar
  45. Massadeh AM, Al-Safi SA, Momani IF, Alomary AA, Jaradat QM, AlKofahi AS (2007) Garlic (Allium sativum L.) as a potential antidote for cadmium and lead intoxication: cadmium and lead distribution and analysis in different mice organs. Biol Trace Elem Res 120:227–234. doi: 10.1007/s12011-007-8017-3 CrossRefPubMedGoogle Scholar
  46. Moreira EG, Rosa GJ, Barros SB, Vassilieff VS, Vassillieff I (2001) Antioxidant defense in rat brain regions after developmental lead exposure. Toxicology 169:145–151CrossRefPubMedGoogle Scholar
  47. Munoz-Pinedo C (2012) Signaling pathways that regulate life and cell death: evolution of apoptosis in the context of self-defense. Adv Exp Med Biol 738:124–143. doi: 10.1007/978-1-4614-1680-7_8 CrossRefPubMedGoogle Scholar
  48. Murphy KJ, Regan CM (1999) Low-level lead exposure in the early postnatal period results in persisting neuroplastic deficits associated with memory consolidation. J Neurochem 72:2099–2104CrossRefPubMedGoogle Scholar
  49. Needleman HL (2004) Low level lead exposure and the development of children. Southeast Asian J Trop Med Public Health 35:252–254PubMedGoogle Scholar
  50. Oberto A, Marks N, Evans HL, Guidotti A (1996) Lead (Pb + 2) promotes apoptosis in newborn rat cerebellar neurons: pathological implications. J Pharmacol Exp Ther 279:435–442PubMedGoogle Scholar
  51. Okada Y, Tanaka K, Fujita I, Sato E, Okajima H (2005) Antioxidant activity of thiosulfinates derived from garlic. Redox Rep 10:96–102. doi: 10.1179/135100005X38851 CrossRefPubMedGoogle Scholar
  52. Olsen RK, Moses SN, Riggs L, Ryan JD (2012) The hippocampus supports multiple cognitive processes through relational binding and comparison. Front Hum Neurosci 6:146. doi: 10.3389/fnhum.2012.00146 CrossRefPubMedPubMedCentralGoogle Scholar
  53. Olympio KP, Goncalves C, Gunther WM, Bechara EJ (2009) Neurotoxicity and aggressiveness triggered by low-level lead in children: a review. Rev Panam Salud Publica 26:266–275CrossRefPubMedGoogle Scholar
  54. Petit TL, LeBoutillier JC (1979) Effects of lead exposure during development on neocortical dendritic and synaptic structure. Exp Neurol 64:482–492CrossRefPubMedGoogle Scholar
  55. Popov I, Blumstein A, Lewin G (1994) Antioxidant effects of aqueous garlic extract. 1st communication: Direct detection using the photochemiluminescence. Arzneimittelforschung 44:602–604PubMedGoogle Scholar
  56. Pourjafar M, Aghbolaghi PA, Shakhse-Niaie M (2007) Effect of garlic along with lead acetate administration on lead burden of some tissues in mice. Pak J Biol Sci 10:2772–2774CrossRefPubMedGoogle Scholar
  57. Prasad K, Laxdal VA, Yu M, Raney BL (1995) Antioxidant activity of allicin, an active principle in garlic. Mol Cell Biochem 148:183–189CrossRefPubMedGoogle Scholar
  58. Privalova LI et al. (2002) [Role of environmental pollution with lead and maintaining psychological development of preschool age children] Vestn Ross Akad Med Nauk:50–53Google Scholar
  59. Rajabzadeh A, Bideskan AE, Fazel A, Sankian M, Rafatpanah H, Haghir H (2012) The effect of PTZ-induced epileptic seizures on hippocampal expression of PSA-NCAM in offspring born to kindled rats. J Biomed Sci 19:56. doi: 10.1186/1423-0127-19-56 CrossRefPubMedPubMedCentralGoogle Scholar
  60. Riexinger KG, Petit TL, Dudek FE (1986) The effects of lead exposure on field potentials of CA3 pyramidal cells from mossy fiber stimulation in rat hippocampus. Neurotoxicology 7:35–45PubMedGoogle Scholar
  61. Robertson JD, Orrenius S (2000) Molecular mechanisms of apoptosis induced by cytotoxic chemicals. Crit Rev Toxicol 30:609–627. doi: 10.1080/10408440008951122 CrossRefPubMedGoogle Scholar
  62. Rogan WJ, Ware JH (2003) Intellectual impairment in children with blood lead concentrations below 10 microg per deciliter. J Pediatr 143:687–688PubMedGoogle Scholar
  63. Ronchetti R, van den Hazel P, Schoeters G, Hanke W, Rennezova Z, Barreto M, Villa MP (2006) Lead neurotoxicity in children: is prenatal exposure more important than postnatal exposure? Acta Paediatr Suppl 95:45–49. doi: 10.1080/08035320600886224 CrossRefPubMedGoogle Scholar
  64. Ruan DY, Yan KF, Ge SY, Xu YZ, Chen JT, Wang M (2000) Effects of chronic lead exposure on short-term and long-term depression in area CA1 of the rat hippocampus in vivo. Chemosphere 41:165–171CrossRefPubMedGoogle Scholar
  65. Schnaas L, Rothenberg SJ, Perroni E, Martinez S, Hernandez C, Hernandez RM (2000) Temporal pattern in the effect of postnatal blood lead level on intellectual development of young children. Neurotoxicol Teratol 22:805–810CrossRefPubMedGoogle Scholar
  66. Schnaas L et al. (2006) Reduced intellectual development in children with prenatal lead exposure. Environ Health Perspect 114:791–797CrossRefPubMedGoogle Scholar
  67. Selvin-Testa A, Lopez-Costa JJ, Nessi de Avinon AC, Pecci Saavedra J (1991) Astroglial alterations in rat hippocampus during chronic lead exposure. Glia 4:384–392. doi: 10.1002/glia.440040406 CrossRefPubMedGoogle Scholar
  68. Senapati SK, Dey S, Dwivedi SK, Swarup D (2001) Effect of garlic (Allium sativum L.) extract on tissue lead level in rats. J Ethnopharmacol 76:229–232CrossRefPubMedGoogle Scholar
  69. Sharifi AM, Baniasadi S, Jorjani M, Rahimi F, Bakhshayesh M (2002) Investigation of acute lead poisoning on apoptosis in rat hippocampus in vivo. Neurosci Lett 329:45–48CrossRefPubMedGoogle Scholar
  70. Shen XM et al. (1992) The adverse effect of marginally higher lead level on intelligence development of children: a Shanghai study. Indian J Pediatr 59:233–238CrossRefPubMedGoogle Scholar
  71. Shen XM et al. (1998) Low-level prenatal lead exposure and neurobehavioral development of children in the first year of life: a prospective study in Shanghai. Environ Res 79:1–8. doi: 10.1006/enrs.1998.3851 CrossRefPubMedGoogle Scholar
  72. Squire LR (1992) Memory and the hippocampus: a synthesis from findings with rats, monkeys, and humans. Psychol Rev 99:195–231CrossRefPubMedGoogle Scholar
  73. Tellez-Rojo MM et al. (2006) Longitudinal associations between blood lead concentrations lower than 10 microg/dL and neurobehavioral development in environmentally exposed children in Mexico City. Pediatrics 118:e323–e330. doi: 10.1542/peds.2005-3123 CrossRefPubMedGoogle Scholar
  74. Thacker SB, Hoffman DA, Smith J, Steinberg K, Zack M (1992) Effect of low-level body burdens of lead on the mental development of children: limitations of meta-analysis in a review of longitudinal data. Arch Environ Health 47:336–346. doi: 10.1080/00039896.1992.9938372 CrossRefPubMedGoogle Scholar
  75. Tomoum HY, Mostafa GA, Ismail NA, Ahmed SM (2010) Lead exposure and its association with pubertal development in school-age Egyptian children: pilot study. Pediatr Int 52:89–93. doi: 10.1111/j.1442-200X.2009.02893.x CrossRefPubMedGoogle Scholar
  76. Verina T, Rohde CA, Guilarte TR (2007) Environmental lead exposure during early life alters granule cell neurogenesis and morphology in the hippocampus of young adult rats. Neuroscience 145:1037–1047. doi: 10.1016/j.neuroscience.2006.12.040 CrossRefPubMedPubMedCentralGoogle Scholar
  77. Walker PR, Sikorska M (1997) New aspects of the mechanism of DNA fragmentation in apoptosis. Biochem Cell Biol 75:287–299CrossRefPubMedGoogle Scholar
  78. Winneke G, Kramer U (1997) Neurobehavioral aspects of lead neurotoxicity in children. Cent Eur J Public Health 5:65–69PubMedGoogle Scholar
  79. Xu J, Ji LD, Xu LH (2006) Lead-induced apoptosis in PC 12 cells: involvement of p53, Bcl-2 family and caspase-3. Toxicol Lett 166:160–167. doi: 10.1016/j.toxlet.2006.06.643 CrossRefPubMedGoogle Scholar
  80. Zailina H, Junidah R, Josephine Y, Jamal HH (2008) The influence of low blood lead concentrations on the cognitive and physical development of primary school children in Malaysia. Asia Pac J Public Health 20:317–326. doi: 10.1177/1010539508322697 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Ali-Reza Ebrahimzadeh-Bideskan
    • 1
  • Javad Hami
    • 2
    • 3
  • Fatemeh Alipour
    • 1
  • Hossein Haghir
    • 1
  • Ali-Reza Fazel
    • 1
  • Akram Sadeghi
    • 4
  1. 1.Department of Anatomy and Cell Biology, School of MedicineMashhad University of Medical SciencesMashhadIran
  2. 2.Department of Anatomical Sciences, School of MedicineBirjand University of Medical SciencesBirjandIran
  3. 3.Cellular and Molecular Research CenterBirjand University of Medical SciencesBirjandIran
  4. 4.Department of Anatomy and Cell Biology, School of MedicineIsfahan University of Medical SciencesIsfahanIran

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