NeuroMolecular Medicine

, Volume 15, Issue 1, pp 192–208 | Cite as

Fisetin Enhances Behavioral Performances and Attenuates Reactive Gliosis and Inflammation During Aluminum Chloride-Induced Neurotoxicity

  • Dharmalingam Prakash
  • Kulasekaran Gopinath
  • Ganapasam Sudhandiran
Original Paper
First Online:
Received:
Accepted:

Abstract

Aluminum (Al) is an environmental neurotoxin that affects cerebral functions and causes health complications. However, the role of Al in arbitrating glia homeostasis and pathophysiology remains obscure. Astrocyte, microglia activation (reactive gliosis), and associated inflammatory events play a decisive role in neurodegeneration and may represent a target for treating neurodegenerative disorders. In this study, we have analyzed the role of aluminum chloride (AlCl3) in causing reactive gliosis in the brain of mice and the ability of fisetin, a flavonoid to attenuate reactive gliosis and neuronal inflammation. Reports suggest that fisetin exerts antioxidant and anti-inflammatory actions. Fisetin at a dose of 15 mg/kg body weight was orally administered, daily (pre-treated for 4 weeks before AlCl3 induction and co-treated until experimental period of 8 weeks) to mice induced with AlCl3 (200 mg/kg b.wt./day/8 weeks, orally). Administration of AlCl3 developed behavioral deficits, triggered lipid peroxidation (LPO), compromised acetylcholine esterase (AChE) activity, and reduced the levels of superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), and reduced glutathione (GSH), and caused histologic aberrations. These effects were accompanied by increased expressions of Glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1. Pro-inflammatory cytokines, such as tumor necrosis factor alpha, interleukin-1β, inducible nitric oxide synthase, were increased upon AlCl3 administration. AlCl3-induced alterations in the activities of SOD, CAT, GST, AChE and levels of GSH, LPO, activity of AChE, behavioral deficits, histologic aberrations, reactive gliosis, and inflammatory niche were attenuated on treatment with fisetin. Collectively, our results indicate that fisetin exerts neuroprotection against AlCl3-induced brain pathology.

Keywords

Fisetin GFAP Iba-1 Reactive gliosis TNF-α IL1-β iNOS Aluminum chloride 
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Notes

Acknowledgments

This work is supported in part by a major grant in neurosciences awarded to GS from Department of Science and Technology (DST), New Delhi, Govt. of India. We thank Dr. Niranjali Devaraj, Professor & Head, Department of Biochemistry, and Dr. H. Devaraj, Professor, Department of Zoology, University of Madras, Guindy campus for their helpful discussions in doctoral committee meeting of DP.

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

12017_2012_8210_MOESM1_ESM.jpg (366 kb)
Supplementary Fig. 1 shows the effect of fisetin on hanging wire test upon AlCl3-induced neuromuscular dysfunction. Graph shows the total time spent in seconds (S) by the animals in the control and experimental groups to hang under the grid. Aluminum chloride (AlC3)-induced mice exhibited significantly low latency to fall in hanging wire test. Treatment with fisetin [pre-treatment (PT) and co-treatment (CT)] showed an improved performance in the hanging wire test. Hypothesis testing method included two-way analysis of variance (ANOVA) followed by post hoc Tukey’s test. Results were expressed as Mean ± SD; given as statistically significant at p<0.05; compared with (a): group 1, (b): group 2, (c): group 3 and, (d): group 4 (JPEG 366kb)
12017_2012_8210_MOESM2_ESM.jpg (1.7 mb)
Supplementary Fig. 2 shows the effect of fisetin on gait performances upon AlCl3-induced behavioral deficits in mice. Front paws were colored with tomato red, and the hind paws were colored with dark green (non-toxic water colors). Representative foot print patterns of control and experimental groups of mice are shown. In particular, three parameters have been recorded. Stride length (cm), sway length (cm), and stance length (cm). Qualitative analyses of foot prints showed significant differences in the stride, sway, and stance lengths. Disoriented walking patterns in Aluminum chloride (AlCl3)-induced mice are noticed form increases stride to stance length, which is typically abnormal as compared with control. Treatment with fisetin [pre-treatment (PT) and co-treatment (CT)] significantly improved the gait performance in AlCl3-induced mice. Hypothesis testing method included two-way analysis of variance (ANOVA) followed by post hoc Tukey’s test. Results were expressed as Mean ± SD; given as statistically significant at p<0.05; compared with (a): group 1, (b): group 2, (c): group 3, and (d): group 4 (JPEG 1711kb)

References

  1. Aremu, D. A., & Meshitsuka, S. (2006). Some aspects of astroglial functions and aluminum implications for neurodegeneration. Brain Research Reviews, 52, 193–200.PubMedCrossRefGoogle Scholar
  2. Ashokkumar, P., & Sudhandiran, G. (2011). Luteolin inhibits cell proliferation during Azoxymethane-induced experimental colon carcinogenesis via Wnt/β-catenin pathway. Investigational New Drugs, 29, 273–284.PubMedCrossRefGoogle Scholar
  3. Bal-Price, A., & Brown, G. C. (2001). Inflammatory neurodegeneration mediated by nitric oxide from activated glia-inhibiting neuronal respiration, causing glutamate release and excitotoxicity. Journal of Neuroscience, 21, 6480–6491.PubMedGoogle Scholar
  4. Bancroft, J. D., & Stevens, A. (Eds.). (1982). Theory and practice of histological techniques (foreword by I. M. P. Dawson) (2nd ed.). Edinburgh, NY: Churchill Livingstone.Google Scholar
  5. Bargagna-Mohan, P., Paranthan, R. R., Hamza, A., Dimova, N., Trucchi, B., Srinivasan, C., et al. (2010). Withaferin A targets intermediate filaments glial fibrillary acidic protein and vimentin in a model of retinal gliosis. Journal of Biological Chemistry, 285, 7657–7669.PubMedCrossRefGoogle Scholar
  6. Bevins, R. A., & Besheer, J. (2006). Object recognition in rats and mice: A one-trial non-matching-to-sample learning task to study ‘recognition memory. Nature Protocols, 1, 11306–11311.CrossRefGoogle Scholar
  7. Carter, R. J., Morton, J., & Dunnett S. B. (2001). Motor coordination and balance in rodents. Current protocols in neuroscience, chap. 8: Unit 8.12.Google Scholar
  8. Chen, T., Hung, H., Wang, D., & Chen, S. S. (2010). The protective effect of Rho-associated kinase inhibitor on aluminum-induced neurotoxicity in rat cortical neurons. Toxicological Sciences, 116, 264–272.PubMedCrossRefGoogle Scholar
  9. Chiruta, C., Schubert, D., Dargusch, R., & Maher, P. (2012). Chemical modification of the multitarget neuroprotective compound fisetin. Journal of Medicinal Chemistry, 55, 378–389.PubMedCrossRefGoogle Scholar
  10. Cho, H., Kim, S., Lee, S., Park, J., Kim, S., & Chun, H. (2008). Protective effect of green tea component, L-theanine on environmental toxins-induced neuronal cell death. Neurotoxicology, 29, 656–662.PubMedCrossRefGoogle Scholar
  11. Cruz, A., Tunez, I., Martinez, R., Munoz-Castaneda, J. R., Ramirez, L. M., Recio, M., et al. (2007). Melatonin prevents brain oxidative stress induced by obstructive jaundice in rats. Journal of Neuroscience Research, 85, 3652–3656.PubMedCrossRefGoogle Scholar
  12. Draper, H. H., Squires, E. J., Mahmoodi, H., Wu, J., Agarwal, S., & Hadley, M. A. (1993). Comparative evaluation of thiobarbituric acid methods for the determination of malondialdehyde in biological materials. Free Radical Biology & Medicine, 15, 353–363.CrossRefGoogle Scholar
  13. Dumont, M. (2011). Behavioral phenotyping of mouse models of neurodegeneration. Methods in Molecular Biology, 793, 229–237.PubMedCrossRefGoogle Scholar
  14. Eckenstein, F., & Sofroniew, M. V. (1983). Identification of central cholinergic neurons containing both choline acetyltransferase and acetylcholinesterase and of central neurons containing only acetylcholinesterase. Journal of Neuroscience, 3, 2286–2291.PubMedGoogle Scholar
  15. Ellman, G. L. (1959). Tissue sulfhydryl groups. Archives of Biochemistry and Biophysics, 82, 70.PubMedCrossRefGoogle Scholar
  16. Ellman, G. L., Courteney, K. D., Andres, V., & Featherstone, R. M. A. (1961). New and rapid colorimetric determination of acetylcholinesterase activity. Biochemical Pharmacology, 7, 88–95.PubMedCrossRefGoogle Scholar
  17. Fukui, K., Onodera, K., Shinkai, T., Suzuki, S., & Urano, S. (2001). Impairment of learning and memory in rats caused by oxidative stress and aging, and changes in antioxidative defense systems. Annals of the New York Academy of Sciences, 928, 168–175.PubMedCrossRefGoogle Scholar
  18. Glowinski, J., & Iversen, L. L. (1996). Regional studies of catecholamines in the rat brain. Journal of Neurochemistry, 13, 655–669.CrossRefGoogle Scholar
  19. Goth, J. (1999). A simple method for determination of serum catalase activity and revision of reference range. Clinica Chimica Acta, 196, 143.CrossRefGoogle Scholar
  20. Guix, F. X., Uribesalgo, M., Coma, M., & Munoz, F. J. (2005). The physiology and pathophysiology of nitric oxide in the brain. Progress in Neurobiology, 76, 126–152.PubMedCrossRefGoogle Scholar
  21. Habig, W. H., & Jakoby, W. B. (1981). Assays for differentiation of glutathione-s-transferases. Methods in Enzymology, 77, 398–405.PubMedCrossRefGoogle Scholar
  22. Haettig, J., Stefanko, D. P., Multani, M. L., Figueroa, D. X., McQuown, S. C., & Wood, M. A. (2011). HDAC inhibition modulates hippocampus-dependent long-term memory for object location in a CBP-dependent manner. Learning and Memory, 18, 71–79.PubMedCrossRefGoogle Scholar
  23. Hald, A., & Lotharius, J. (2005). Oxidative stress and inflammation in Parkinson’s disease: Is there a causal link? Experimental Neurology, 193, 279–290.PubMedCrossRefGoogle Scholar
  24. Heneka, M. T., & O’Banion, M. K. (2007). Inflammatory processes in Alzheimer’s disease. Journal of Neuroimmunology, 184, 69–91.PubMedCrossRefGoogle Scholar
  25. Hong, J., Cho, I., Kwak, K. I., Cheng Suh, E., Seo, J., Min, H. J., et al. (2010). Microglial toll-like receptor 2 contributes to kainic acid-induced glial activation and hippocampal neuronal cell death. Journal of Biological Chemistry, 50, 39447–39457.CrossRefGoogle Scholar
  26. Ikeda, Y., Hayashi, M., Dohi, K., & Matsumoto, K. (2001). Biochemical markers for brain damage. Neurosurgery Quarterly, 11, 173–180.CrossRefGoogle Scholar
  27. Kaushik, D. K., Mukhopadhyay, R., Kumawat, K. L., Gupta, M., & Basu, A. (2012). Therapeutic targeting of Krüppel-like factor 4 abrogates microglial activation. Journal of Neuroinflammation, 9, 57.PubMedCrossRefGoogle Scholar
  28. Kleine, T. O., Benes, L., & Zofel, P. (2003). Studies of the brain specificity of S100B and neuron-specific enolase (NSE) in blood serum of acute care patients. Brain Research Bulletin, 61, 265–279.PubMedCrossRefGoogle Scholar
  29. Kono, Y. (1978). Generation of superoxide radical during auto-oxidation of hydroxylamine and an assay of superoxide dismutase. Archives of Biochemistry Biophysics, 186, 189–195.CrossRefGoogle Scholar
  30. Lowry, O. H., Rosenbrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193, 265–275.PubMedGoogle Scholar
  31. Maher, P. (2009). Modulation of multiple pathways involved in the maintenance of neuronal function during aging by fisetin. Genes and Nutrition, 4, 297–307.PubMedCrossRefGoogle Scholar
  32. Maher, P., Salgado, K. F., Zivin, J. A., & Lapchak, P. A. (2007). A novel approach to screening for new neuroprotective compounds for the treatment of stroke. Brain Research, 1173, 117–125.PubMedCrossRefGoogle Scholar
  33. Montezumaa, K., Caroline Biojoneb, C., Lisboab, S. F., Cunhab, F. Q., Guimaraes, F. S., & Jocaa, S. R. L. (2012). Inhibition of iNOS induces antidepressant-like effects in mice: Pharmacological and genetic evidence. Neuropharmacology, 62, 485–491.CrossRefGoogle Scholar
  34. O’Callaghan, J. P., Sriram, K., & Miller, D. B. (2008). Defining neuroinflammation. Annals of the New York Academy of Sciences, 1139, 318–330.PubMedCrossRefGoogle Scholar
  35. Ohsawa, K., Imai, Y., Kanazawa, H., Sasaki, Y., & Kohsaka, S. (2000). Involvement of Iba-1 in membrane ruffling and phagocytosis of macrophages/microglia. Journal of Cell Science, 113, 3073–3084.PubMedGoogle Scholar
  36. Papandreoua, M. A., Tsachaki, M., Efthimiopoulos, S., Cordopatisc, P., Lamaric, F. N., & Margarity, M. (2011). Memory enhancing effects of saffron in aged mice are correlated with antioxidant protection. Behavioural Brain Research, 219, 197–204.CrossRefGoogle Scholar
  37. Park, K. M., & Bowers, W. J. (2010). Tumor necrosis factor-alpha mediated signaling in neuronal homeostasis and dysfunction. Cellular Signalling, 22, 977–983.PubMedCrossRefGoogle Scholar
  38. Petrik, S. M., Wong, C. M., Tabata, C. R., Garry, F. R., & Shaw, C. A. (2007). Aluminum adjuvant linked to Gulf war illness induces motor neuron death in mice. NeuroMolecular Medicine, 9, 83–100.PubMedCrossRefGoogle Scholar
  39. Prut, L., & Belzung, C. (2003). The open field as a paradigm to measure the effects of drugs on anxiety-like behaviors: A review. European Journal of Pharmacology, 463, 3–33.PubMedCrossRefGoogle Scholar
  40. Reale, M., Larlori, C., Thomas, A., Gambi, D., Perfetti, B., Di Nicola, M., et al. (2009). Peripheral cytokines profile in Parkinson’s disease. Brain Behavioural Immunity, 23, 55–63.CrossRefGoogle Scholar
  41. Rendeiro, C., Guerreiro, J. D., Williams, C. M., & Spencer, J. P. (2012). Flavonoids as modulators of memory and learning: Molecular interactions resulting in behavioural effects. Proceedings of the Nutrition Society, 71, 246–262.PubMedCrossRefGoogle Scholar
  42. Rolls, A., Shechter, R., & Schwartz, M. (2009). The bright side of the glial scar in CNS repairs. Nature Review Neuroscience, 10, 235–241.CrossRefGoogle Scholar
  43. Rothwell, N. J., & Luheshi, G. N. (2000). Interleukin 1 in the brain: Biology, pathology, and therapeutic target. Trends in Neuroscience, 23, 618–625.CrossRefGoogle Scholar
  44. Rui, D., & Yongjian, Y. (2010). Aluminum chloride induced oxidative damage on cells derived from hippocampus and cortex of ICR mice. Brain Research, 1324, 96–102.PubMedCrossRefGoogle Scholar
  45. Saha, R. N., & Pahan, K. (2006). Regulation of inducible nitric oxide synthase gene in glial cells. Antioxidants & Redox Signaling, 8, 929–947.CrossRefGoogle Scholar
  46. Santosa, D., Milatovic, D., Andradea, V., Batoreua, C., Aschner, M., & Marreilha dos Santosa, A. P. (2012). The inhibitory effect of manganese on acetylcholinesterase activity enhances oxidative stress and neuroinflammation in the rat brain. Toxicology, 292, 90–98.CrossRefGoogle Scholar
  47. Savory, J., Herman, M. M., & Ghribi, O. (2006). Mechanisms of aluminum-induced neurodegeneration in animals: Implications for Alzheimer’s disease. Journal of Alzheimer’s Disease, 10, 135–144.PubMedGoogle Scholar
  48. Shaw, C. A., & Petrik, M. S. (2009). Aluminum hydroxide injections lead to motor deficits and motor neuron degeneration. Journal of Inorganic Biochemistry, 103, 1555–1562.PubMedCrossRefGoogle Scholar
  49. Silman, I., & Sussman, J. (2005). Acetylcholinesterase: Classical and non-classical functions and pharmacology. Current Opinion in Pharmacology, 5, 293–302.PubMedCrossRefGoogle Scholar
  50. Tan, Z. S., Beiser, A. S., Vasan, R. S., Roubenoff, R., Dinarello, C. A., Harris, T. B., et al. (2007). Inflammatory markers and the risk of Alzheimer disease: The Framingham Study. Neurology, 68, 1902–1908.PubMedCrossRefGoogle Scholar
  51. Teismann, P., & Schulz, J. B. (2004). Cellular pathology of Parkinson’s disease: Astrocytes, microglia and inflammation. Cell Tissue Research, 318, 149–161.PubMedCrossRefGoogle Scholar
  52. Wang, Y., Wu, Y., Li, L., Zheng, J., Liu, R., Zhou, J., et al. (2011). Aspirin-triggered lipoxin A4 attenuates LPS-induced pro-inflammatory responses by inhibiting activation of NF-κB and MAPKs in BV-2 microglial cells. Journal of Neuroinflammation, 8, 95.PubMedCrossRefGoogle Scholar
  53. Zatta, P., Ibn-Lkhayat, M., Zambenedetti, P., Kilyen, M., & Kiss, T. (2002). In vivo and In vitro effects of aluminium on the activity of mouse brain acetylcholine esterase. Brain Research Bulletin, 59, 41–45.PubMedCrossRefGoogle Scholar
  54. Zeng, K., Fu, H., Liu, G., & Wang, X. (2012). Aluminum maltolate induces primary rat astrocyte apoptosis via overactivation of the class III PI3 K/Beclin 1-dependent autophagy signal. Toxicology in Vitro, 26, 215–220.PubMedCrossRefGoogle Scholar
  55. Zhang, Q. G., Laird, M. D., Han, D., Nguyen, K., Scott, E., Dong, Y., et al. (2012). Critical role of NADPH oxidase in neuronal oxidative damage and microglia activation following traumatic brain injury. PLoS ONE, 7, e34504.PubMedCrossRefGoogle Scholar
  56. Zhao, F., Cai, T., Liu, M., Zheng, G., Luo, W., & Chen, J. (2009). Manganese induces dopaminergic neurodegeneration via microglial activation in a rat model of manganism. Toxicological Sciences, 107, 156–164.PubMedCrossRefGoogle Scholar
  57. Zheng, L. T., Ock, J., Kwon, B. M., & Suk, K. (2008). Suppressive effects of flavonoid fisetin on lipopolysaccharide-induced microglial activation and neurotoxicity. International Immunopharmacology, 8, 484–494.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Dharmalingam Prakash
    • 1
  • Kulasekaran Gopinath
    • 1
  • Ganapasam Sudhandiran
    • 1
  1. 1.Department of Biochemistry, Cell Biology LaboratoryUniversity of MadrasChennaiIndia

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