Neurochemical Research

, Volume 40, Issue 8, pp 1699–1708 | Cite as

Neuroprotective Role of Lazaroids Against Aluminium Chloride Poisoning

  • Pooja Khanna Sood
  • Sonia Verma
  • Uma Nahar
  • Bimla Nehru
Original Paper


Aluminium (Al) is neurotoxic primarily because of its interference with biological enzymes in key mechanisms of metabolic pathways. Mitochondria being a major site of reactive oxygen species (ROS) production, it seems that the oxidative damage to mitochondrial proteins may underlie the pathogenesis of Al induced neurodegeneration. The present study investigates the effectiveness of the anti-oxidant property of lazaroids (U-74500A), a known lipid peroxidation inhibitor as neuroprotective agent against Al induced neurotoxicity. Al chloride was administered orally at a dose level of 100 mg/kg body wt/day in water and U-74500A was administered at a dose of 0.25 mg/kg body wt i.p. in citrate buffer for a period of 8 weeks on alternate days. Following Al exposure there was a significant increase in lipid peroxidation (LPO), ROS levels and reduction in the activity of mitochondrial complexes in all the three regions of rat brain, i.e., cerebral cortex, mid brain, and cerebellum. This decrease in the activities of electron transport complexes in turn affected the ATP synthesis and ATP levels adversely in the mitochondria. These alterations were also depicted in the histology which shows signs of hypoxia, paucity of neurons in cortical region and loosening of fibers in the white matter. U-74500A co-administration was able to restore alterations in the LPO, ROS levels as well as all the three mitochondrial complexes and caspase expression. Therefore, it is suggested that 21-aminosteroids (lazaroids), by attenuating LPO and mitochondrial dysfunction, holds a promise as an agent that can potentially reduce Al-induced adverse effects in brain.


Aluminium neurotoxicity Lipid peroxidation Oxidative phosphorylation Caspase expression Histopathology Lazaroids 





Adenosine 5′diphosphate


Adenosine triphosphate


Bovine serum albumin


Central nervous system




2,7 Dichlorofluorescein


Deoxyribose nucleic acid


Ethylene diamine tricholoacetic acid


Ethylene glycol tetraacetic acid


Electron transport chain






Potassium chloride


Potassium dihydrogen phosphate




Lipid peroxidation




Magnesium chloride


(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-H-tetrazolium bromide


Mitochondrial transition permeability pore


Nicotinamide adenine dinucleotide reduced


Nicotinamide adenine dinucleotide phosphate reduced




Inorganic phosphate




Post mitochondrial fraction


Ribose nucleic acid


Reactive oxygen species


Sodium dodecyl sulphate


Sodium dodecyl sulphate poly acrylamide gel electrophoresis

Tris HCl

Tris hydrochloride


Thiobarbituric acid


Tricholro acetic acid



Financial support from ICMR in the form of Research fellowship to Ms Pooja Khanna is highly appreciated.

Compliance with Ethical Standards

Conflict of interest

The authors don’t have any conflicting interest in the manuscript.

Ethical standard

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were approved by Institutional Animal Ethics Committee (IAEC), Panjab University, Chandigarh, India and in general according to the NIH guidelines (Rule No. 23-85, as revised in 1985).

Human rights

This article does not contain any studies with human participants performed by any of the authors.


  1. 1.
    Colomina MT, Roig JL, Sanchez DL, Domingo JL (2002) Influence of age on Al induced neurobehavioural effects of changes in rat brain. Neurotoxicology 23:775–781CrossRefPubMedGoogle Scholar
  2. 2.
    Rhein V, Baysang G, Rao S et al (2009) Amyloid-beta leads to impaired cellular respiration, energy production and mitochondrial electron chain complex activities in human neuroblastoma cells. Cell Mol Neurobiol 29:1063–1071CrossRefPubMedGoogle Scholar
  3. 3.
    Exley C (2004) The pro-oxidant activity of Al. Free Radic Biol Med 36:380–387CrossRefPubMedGoogle Scholar
  4. 4.
    Kaneko N, Sugioka T, Sakurai H (2007) Al compounds enhance LPO in liposomes: insight into cellular damage caused by oxidative stress. J Inorg Biochem 101:967–975CrossRefPubMedGoogle Scholar
  5. 5.
    Grundke-Iqbal I, Wang GP, Iqbal K, Wisniewski HM (1985) Alzheimer paired helical filaments: identification of polypeptides with monoclonal antibodies. Acta Neuropathol 68:279–283CrossRefPubMedGoogle Scholar
  6. 6.
    Jacobsen EJ, Mccall JM, Ayer DE, Vandoornik FJ et al (1990) Novel 21 aminosteroids that inhibit iron-dependent lipid peroxidation and protect against central nervous system trauma. J Med Chem 33:1145–1151CrossRefPubMedGoogle Scholar
  7. 7.
    Fukuma K, Marubayashi S, Okada K, Yamada K et al (2009) Effect of lazaroid U-74389G and methylprednisolone on endotoxin-induced shock in mice. Surgery 125:421–430CrossRefGoogle Scholar
  8. 8.
    Vignes JR, Hugon J (2006) In vitro efficacy of three lazaroids in a model of acute chemical neuronal hypoxia. Neurosci Lett 407:171–175CrossRefPubMedGoogle Scholar
  9. 9.
    Villa RF, Gorini A (1997) Pharmacology of lazaroids and brain energy metabolism: A review. Pharmacol Rev 49(1):99–136PubMedGoogle Scholar
  10. 10.
    Sood PK, Nahar U, Nehru B (2012) Stress proteins and glial cell functions during chronic Al exposures: protective role of curcumin. Neurochem Res 37:639–646CrossRefPubMedGoogle Scholar
  11. 11.
    Bishnoi M, Chopra K, Kulkarni SK (2007) U-74500A (lazaroid), a 21-aminosteroid attenuates neuroleptic-induced orofacial dyskinesia. Methods Find Exp Clin Pharmacol 29(9):601–605CrossRefPubMedGoogle Scholar
  12. 12.
    Wills ED (1966) Mechanism of lipid peroxide formation in animal tissues. Biochem J 99:667–676PubMedCentralCrossRefPubMedGoogle Scholar
  13. 13.
    Berman SB, Hastings TG (1999) Dopamine oxidation alters mitochondrial respiration and induces permeability transition in brain mitochondria: implications for Parkinson’s disease. J Neurochem 73:1127–1137CrossRefPubMedGoogle Scholar
  14. 14.
    King TE, Howard RL (1967) Preparation and properties of soluble NADH dehydrogenase from cardiac muscle. In: Colowick SP, Kaplan NO (eds) Methods in enzymology. Academic Press, New York, pp 275–276Google Scholar
  15. 15.
    King TS (1967) Preparation of succinate dehydrogenase and reconstitution of succinate oxidase. Methods in enzymology. Academic Press, New York, pp 322–325Google Scholar
  16. 16.
    Sandhir R, Mehrotra A, Kamboj SS (2010) Lycopene prevents 3-nitropropionic acid-induced mitochondrial oxidative stress and dysfunctions in nervous system. Neurochem Int 57:579–587CrossRefPubMedGoogle Scholar
  17. 17.
    Griffiths DE, Cain K, Hyams RL (1977) Studies on energy linked reactions: inhibition of oxidative phosphorylation by DL-8-methyl dihydrolipoate. Biochem J 164:699–704PubMedCentralCrossRefPubMedGoogle Scholar
  18. 18.
    Griffiths DE, Houghton RL (1974) Studies on energy linked reactions: modified mitochondrial ATPase of oligomycin-resistant mutants of saccharomyces cerevisiae. Eur J Biochem 46:157–167CrossRefPubMedGoogle Scholar
  19. 19.
    Driver AS, Kodavanti PR, Mundy WR (2000) Age- related changes in reactive oxygen species production in rat brain homogenates. Neurotoxicol Teratol 22:175–181CrossRefPubMedGoogle Scholar
  20. 20.
    Pearse AGE (1968) Histochemistry, theoretical and applied, vol 1; 3rd edn. Churchill Livingstone, London, p 660Google Scholar
  21. 21.
    Humanson GL (1961) Basic procedures—animal tissue technique, vol.1, pp 130–132Google Scholar
  22. 22.
    Lowry OH, Rosebrough NJ, Farr AL, Ranell RJ (1951) Protein measurements with the Follin’s phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  23. 23.
    Jarskog LF, John HG, Leisa AG, Karissa LG et al (2007) Caspase-3 activation in rat frontal cortex following treatment with typical and atypical antipsychotics. Neuropsychopharmacology 32:95–102CrossRefPubMedGoogle Scholar
  24. 24.
    Yuan J, Yanker BA (2000) Apoptosis in the nervous system. Nature 407:802–809CrossRefPubMedGoogle Scholar
  25. 25.
    Arends MJ, Morris RG, Wylie AH (1990) Apoptosis: the role of the endonuclease. Am J Pathol 136:593–608PubMedCentralPubMedGoogle Scholar
  26. 26.
    Smith SL, Scherch HM, Hall ED (1996) Protective effect of tirilazad mesylate and metabolite U-89678 against blood brain barrier damage after subarachnoid hemorrhage and lipid peroxidative neuronal injury. J Neurosurg 84:229–233CrossRefPubMedGoogle Scholar
  27. 27.
    Hinzmann JS, Mckenna RL, Ckenna RL, Pierson TS et al (1992) Interaction of antioxidants with depth-dependent fluorescence quenchers and energy transfer probes in lipid bilayers. Chem Phys Lipids 62:123–138CrossRefPubMedGoogle Scholar
  28. 28.
    Schmid E, Stefan Z, Edwin H, Nikolaus P et al (1997) Superior neuroprotective efficacy of a novel antioxidant (U-101033E) with improved blood-brain barrier permeability in focal cerebral ischemia. Stroke 28:2018–2024CrossRefGoogle Scholar
  29. 29.
    Durmaz R, Kanbak G, Akyuz F, Isiksoy S et al (2003) Lazaroid attenuates edema by stabilizing ATPase in the traumatized rat brain. Can J Neurol Sci 30:143–149PubMedGoogle Scholar
  30. 30.
    Maes M, Fisar Z, Medina M, Scapagnini G et al (2012) New drug targets in depression: inflammatory, cell-mediated immune, oxidative and nitrosative stress, mitochondrial, antioxidant, and neuroprogressive pathways. And new drug candidates—Nrf2 activators and GSK-3 inhibitors. Inflammopharmacology 20:127–150CrossRefPubMedGoogle Scholar
  31. 31.
    Starkov A (2008) The role of mitochondria in reactive oxygen species metabolism and signaling. Ann N Y Acad Sci 1147:37–52PubMedCentralCrossRefPubMedGoogle Scholar
  32. 32.
    Hroudova J, Singh N, Fisar Z (2014) BioMed Res Int. Article ID 175062, 9. doi: 10.1155/175062
  33. 33.
    Carpenter MB (1991) Textbook of neuroanatomy, 4th edn. Williams & Wilkins, BaltimoreGoogle Scholar
  34. 34.
    Saraste M (1999) Oxidative phosphorylation at the fin de siecle. Science 283:1488–1493CrossRefPubMedGoogle Scholar
  35. 35.
    Brown GC (1999) Nitric oxide and mitochondrial respiration. Biochim Biophys Acta 1411:351–369CrossRefPubMedGoogle Scholar
  36. 36.
    Cassina A, Radi R (1996) Differential inhibitory action of nitric oxide and peroxynitrite on mitochondrial electron transport. Arch Biochem Biophys 328:309–316CrossRefPubMedGoogle Scholar
  37. 37.
    Cleeter MW (1994) Reversible inhibition of cytochrome c oxidase, the terminal enzyme of the mitochondrial respiratory chain, by nitric oxide implications for neurodegenerative diseases. FEBS Lett 345:50–54CrossRefPubMedGoogle Scholar
  38. 38.
    Ohyashiki T, Satoh E, Okada M, Takadera T et al (2002) Nerve growth factor protects against Al-mediated cell death. Toxicology 176:195–207CrossRefPubMedGoogle Scholar
  39. 39.
    Mailloux R, Lemire J, Appanna V (2007) Al-induced mitochondrial dysfunction leads to lipid accumulation in human hepatocytes: a link to obesity. Cell Physiol Biochem 20:627–638CrossRefPubMedGoogle Scholar
  40. 40.
    Kraus-Friedman N, Bibes J, Muren H, Carafoli E (1982) Calcium uptake in isolated hepatic plasma membrane vesicles. Eur J Biochem 129:7–12CrossRefGoogle Scholar
  41. 41.
    Bosetti F, Solaini G, Tendi EA, Chikhale EG et al (2001) Mitochondrial cytochrome c oxidase subunit III is selectively down-regulated by Al exposure in PC12S cells. NeuroReport 12:721–724CrossRefPubMedGoogle Scholar
  42. 42.
    Ghribi O, Herman MM, Forbes MS et al (2001) GDNF protects against aluminum-induced apoptosis in rabbits by upregulating Bcl-2 and Bcl-XL and inhibiting mitochondrial Bax translocation. Neurobiol Dis 8:764–773CrossRefPubMedGoogle Scholar
  43. 43.
    Dewitt DA, Hurd JA, Fox N, Townsend BE et al (2006) Peri-nuclear clustering of mitochondria is triggered during Al maltolate induced apoptosis. J Alzheimer Dis 9:195–205Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Pooja Khanna Sood
    • 1
  • Sonia Verma
    • 1
  • Uma Nahar
    • 2
  • Bimla Nehru
    • 1
  1. 1.Biophysics DepartmentPanjab UniversityChandigarhIndia
  2. 2.Histopathology DepartmentPost Graduate Institute of Medial Education and ResearchChandigarhIndia

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