Abstract
Betalains obtained from Beta vulgaris (family Caryophyllales) are regularly consumed as part of the regular diet with medicinal benefits due to antioxidant and anti-inflammatory properties. The objective of this article was to evaluate betanin’s neuroprotective properties in a scopolamine-induced zebrafish paradigm. Betanin (BET) (50, 100, and 200 mg/L), and donepezil (10 mg/L) were delivered to zebrafish in a treatment tank once a day for 8 days, while memory impairment was produced by scopolamine (100 µM), which was given 60 min before behavioral assessments. The treatment dosages were determined based on acute toxicity studies. The existence of betacyanin and betaxanthins of BET was tested using liquid chromatography-mass spectrometry (LC–MS). The Y-maze task was used to examine the novelty and spatial memory, while the novel tank diving test was used to assess anxiety-like behavior (NTT). The activities of acetylcholinesterase (AChE) and the oxidative stress sensitivity in zebrafish brains were examined. Also, brain-derived neurotrophic factor (BDNF) level is quantified by an ELISA kit. Scopolamine-induced rises in AChE activity, memory loss, anxiety, and brain oxidant capacity were all reduced by BET. These results suggest that BET (50 and 100 mg/L) has a therapeutic ability to treat brain oxidative stress and cognitive deficits in amnesic zebrafish.
Graphical Abstract
Similar content being viewed by others
Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author upon reasonable request.
Code availability
NA.
References
De Abreu MS, Friend AJ, Amstislavskaya TG, Kalueff AV (2018) Commentary establishing zebrafish as a model to study the anxiolytic effects of scopolamine. Front Pharmacol 9:293
Baek SY, Li FY, Kim DH et al (2020) Enteromorpha prolifera extract improves memory in scopolamine-treated mice via downregulating amyloid-β expression and upregulating BDNF/TrkB pathway. Antioxidants 9:1–16. https://doi.org/10.3390/antiox9070620
Bastos EL, Schliemann W (2021) Betalains as antioxidants. Plant antioxidants and health. Springer, Cham, pp 1–44
Bekinschtein P, Cammarota M, Igaz LM et al (2007) Persistence of long-term memory storage requires a late protein synthesis- and BDNF- dependent phase in the hippocampus. Neuron 53:261–277. https://doi.org/10.1016/j.neuron.2006.11.025
Bhuvanendran S, Kumari Y, Othman I, Shaikh MF (2018) Amelioration of cognitive deficit by embelin in a scopolamine-induced Alzheimer’s disease-like condition in a rat model. Front Pharmacol 9:1–12. https://doi.org/10.3389/fphar.2018.00665
Boiangiu RS, Mihasan M, Gorgan DL et al (2021) Anxiolytic, promnesic, anti-acetylcholinesterase and antioxidant effects of cotinine and 6-hydroxy-l-nicotine in scopolamine-induced zebrafish (Danio rerio) model of Alzheimer’s disease. Antioxidants 10:1–28. https://doi.org/10.3390/antiox10020212
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254. https://doi.org/10.1016/0003-2697(76)90527-3
Braida D, Ponzoni L, Martucci R et al (2014) Role of neuronal nicotinic acetylcholine receptors (nAChRs) on learning and memory in zebrafish. Psychopharmacology 231:1975–1985. https://doi.org/10.1007/s00213-013-3340-1
Brinza I, Ayoub IM, Eldahshan OA, Hritcu L (2021a) Baicalein 5,6-dimethyl ether prevents memory deficits in the scopolamine zebrafish model by regulating cholinergic and antioxidant systems. Plants 10:1–15. https://doi.org/10.3390/plants10061245
Brinza I, Ayoub IM, Eldahshan OA, Hritcu L (2021b) Baicalein 5 6-dimethyl ether prevents memory deficits in the scopolamine zebrafish model by regulating cholinergic and antioxidant systems. Plants 10. https://doi.org/10.3390/plants10061245
Capatina L, Todirascu-Ciornea E, Napoli EM et al (2020) Thymus vulgaris essential oil protects zebrafish against cognitive dysfunction by regulating cholinergic and antioxidants systems. Antioxidants 9:1–18. https://doi.org/10.3390/antiox9111083
Chen BH, Park JH, Lee TK et al (2018) Melatonin attenuates scopolamine-induced cognitive impairment via protecting against demyelination through BDNF-TrkB signaling in the mouse dentate gyrus. Chem Biol Interact 285:8–13. https://doi.org/10.1016/j.cbi.2018.02.023
Corpuz HM, Fujii H, Nakamura S, Katayama S (2019) Fermented rice peptides attenuate scopolamine-induced memory impairment in mice by regulating neurotrophic signaling pathways in the hippocampus. Brain Res 1720:146322. https://doi.org/10.1016/j.brainres.2019.146322
Da Penha BM, Cooper J, Castren E et al (1993) Cholinergic regulation of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) but not neurotrophin-3 (NT-3) mRNA levels in the developing rat hippocampus. J Neurosci 13:3818–3826. https://doi.org/10.1523/jneurosci.13-09-03818.1993
da Silva Aleluia GA, Keita H, da Silva HR et al (2018) Leaves of Spondias mombin L. a traditional anxiolytic and antidepressant: pharmacological evaluation on zebrafish (Danio rerio). J Ethnopharmacol 224:563–578. https://doi.org/10.1016/j.jep.2018.05.037
Deiana S, Platt B, Riedel G (2011) The cholinergic system and spatial learning. Behav Brain Res 221:389–411. https://doi.org/10.1016/j.bbr.2010.11.036
Desseva I, Stoyanova M, Petkova N, Mihaylova D (2020) Red beetroot juice phytochemicals bioaccessibility an in vitro approach. Polish J Food Nutr Sci 70(45):53. https://doi.org/10.31883/pjfns/116590
Ellman GL, Courtney KD, Andres V, Featherstone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95. https://doi.org/10.1016/0006-2952(61)90145-9
Gulay H, Senturk A, Ince I, Alver A (2016) Assessment of oxidative stress parameters of brain-derived neurotrophic factor heterozygous mice in acute stress model assessment of oxidative stress parameters of brain-derived neurotrophic factor heterozygous mi. Iran J Basic Med Sci 19:388–393
Hamilton TJ, Morrill A, Lucas K et al (2017) Establishing zebrafish as a model to study the anxiolytic effects of scopolamine. Sci Rep 7:1–9. https://doi.org/10.1038/s41598-017-15374-w
Hammond TC, Xing X, Wang C et al (2020) β-amyloid and tau drive early Alzheimer’s disease decline while glucose hypometabolism drives late decline. Commun Biol 31(3):1–13. https://doi.org/10.1038/s42003-020-1079-x
Holcombe A, Schalomon M, Hamilton TJ (2014) A novel method of drug administration to multiple zebrafish Danio rerio and the quantification of withdrawal. J Vis Exp e51851. https://doi.org/10.3791/51851
Howe K, Clark MD, Torroja CF et al (2013) The zebrafish reference genome sequence and its relationship to the human genome. Nature 496:498–503. https://doi.org/10.1038/nature12111
Ibrahim AM, Chauhan L, Bhardwaj A et al (2022) Brain-derived neurotropic factor in neurodegenerative disorders. Biomedicines 10. https://doi.org/10.3390/biomedicines10051143
Karnik I, Gerlai R (2012) Can zebrafish learn spatial tasks? An empirical analysis of place and single CS-US associative learning. Behav Brain Res 233:415–421. https://doi.org/10.1016/j.bbr.2012.05.024
Kaur G, Thawkar B, Dubey S, Jadhav P (2018) Pharmacological potentials of betalains. J Complement Integr Med 15:1–9. https://doi.org/10.1515/jcim-2017-0063
Klinkenberg I, Blokland A (2010) The validity of scopolamine as a pharmacological model for cognitive impairment: a review of animal behavioral studies. Neurosci Biobehav Rev 34:1307–1350
Ko YH, Kwon SH, Lee SY, Jang CG (2017) Liquiritigenin ameliorates memory and cognitive impairment through cholinergic and BDNF pathways in the mouse hippocampus. Arch Pharm Res 40:1209–1217. https://doi.org/10.1007/s12272-017-0954-6
Kruger NJ (1994) The Bradford method for protein quantitation. Methods Mol Biol 32:9–15
Lee JS, Kim HG, Lee HW et al (2015) Hippocampal memory enhancing activity of pine needle extract against scopolamine-induced amnesia in a mouse model. Sci Rep 5:1–10. https://doi.org/10.1038/srep09651
Lian B, Gu J, Zhang C et al (2022) Protective effects of isofraxidin against scopolamine-induced cognitive and memory impairments in mice involve modulation of the BDNF CREB ERK signaling pathway. Metab Brain Dis:1–22. https://doi.org/10.1007/s11011-022-00980-z
Marcus DL, Thomas C, Rodriguez C et al (1998) Increased peroxidation and reduced antioxidant enzyme activity in Alzheimer’s disease. Exp Neurol 150:40–44. https://doi.org/10.1006/exnr.1997.6750
Meshalkina DA, Kizlyk MN, Kysil EV et al (2017) Understanding zebrafish cognition. Behav Processes 141:229–241
Stewart AM, Kalueff AV (2012) The developing utility of zebrafish models for cognitive enhancers research. Curr Neuropharmacol 10:263–271. https://doi.org/10.2174/157015912803217323
Mufson EJ, Counts SE, Perez SE, Ginsberg SD (2008) Cholinergic system during the progression of Alzheimer’s disease: therapeutic implications. Expert Rev Neurother 8:1703–1718. https://doi.org/10.1586/14737175.8.11.1703
OECD Guidelines O, Development Economic Cooperation and 1992 Fish Acute Toxicity Test OECD 203 Effects on biotic systems pdf
Otálora MC, Carriazo JG, Iturriaga L et al (2015) Microencapsulation of betalains obtained from cactus fruit (Opuntia ficus-indica) by spray drying using cactus cladode mucilage and maltodextrin as encapsulating agents. Food Chem 187:174–181. https://doi.org/10.1016/j.foodchem.2015.04.090
Rahimi P, Mesbah-Namin SA, Ostadrahimi A et al (2019) Betalain-and betacyanin-rich supplements impacts on the PBMC SIRT1 and LOX1 genes expression and Sirtuin-1 protein levels in coronary artery disease patients a pilot crossover clinical trial. J Funct Foods 60 103401. https://doi.org/10.1016/j.jff.2019.06.003
Rahnama S, Rabiei Z, Alibabaei Z et al (2015) Anti-amnesic activity of citrus aurantium flowers extract against scopolamine-induced memory impairments in rats. Neurol Sci 36:553–560. https://doi.org/10.1007/s10072-014-1991-2
Rehman S, Ashfaq UA, Sufyan M et al (2022) The insight of in silico and in vitro evaluation of beta vulgaris phytochemicals against Alzheimer’s disease targeting acetylcholinesterase. PLoS ONE 17:1–19. https://doi.org/10.1371/journal.pone.0264074
Richetti SK, Rosemberg DB, Ventura-Lima J et al (2011) Acetylcholinesterase activity and antioxidant capacity of zebrafish brain is altered by heavy metal exposure. Neurotoxicology 32:116–122. https://doi.org/10.1016/j.neuro.2010.11.001
Saleem S, Kannan RR (2018) Zebrafish: an emerging real-time model system to study Alzheimer’s disease and neurospecific drug discovery. Cell Death Discov 4:45. https://doi.org/10.1038/s41420-018-0109-7
Salimi A, Sabur M, Dadkhah M, Shabani M (2022) Inhibition of scopolamine induced memory and mitochondrial impairment by betanin. J Biochem Mol Toxicol 36:e23076. https://doi.org/10.1002/jbt.23076
Spinelli S, Ballard T, Feldon J et al (2006) Enhancing effects of nicotine and impairing effects of scopolamine on distinct aspects of performance in computerized attention and working memory tasks in marmoset monkeys. Neuropharmacology 51:238–250. https://doi.org/10.1016/j.neuropharm.2006.03.012
Squire LR (1992) Memory and the hippocampus: a synthesis from findings with rats, monkeys, and humans. Psychol Rev 99:195–231. https://doi.org/10.1037/0033-295X.99.2.195
Suryavanshi SV, Barve K, Addepalli V et al (2021) Triphala Churna—a traditional formulation in Ayurveda mitigates diabetic neuropathy in rats. Front Pharmacol 12:1–10. https://doi.org/10.3389/fphar.2021.662000
Tan HM, Wills TJ, Cacucci F (2017) The development of spatial and memory circuits in the rat. Wiley Interdiscip Rev Cogn Sci 8
Tan ML, Hamid SBS (2021) Beetroot as a potential functional food for cancer chemoprevention a narrative review. J Cancer Prev 26(1):17. https://doi.org/10.15430/jcp.2021.26.1.1
Thawkar BS, Banerjee M, Kaur G (2023) Alzheimer’s disease preliminary screening in zebrafish integrating behavioral models and molecular markers. In Handbook of animal models in neurological disorders Academic Press, pp 3–16
Thawkar BS, Kaur G (2021) Zebrafish as a promising tool for modeling neurotoxin-induced Alzheimer’s disease. Neurotox Res 39:949–965. https://doi.org/10.1007/s12640-021-00343-z
Valu M-V, Soare LC, Ducu C et al (2021) Hericium erinaceus (Bull.) Pers. ethanolic extract with antioxidant properties on scopolamine-induced memory deficits in a zebrafish model of cognitive impairment. J Fungi 7:477. https://doi.org/10.3390/jof7060477
Volgin AD, Yakovlev OA, Demin KA et al (2019) Acute behavioral effects of deliriant hallucinogens atropine and scopolamine in adult zebrafish. Behav Brain Res 359:274–280. https://doi.org/10.1016/j.bbr.2018.10.033
Zhang L, Fang Y, Xu Y et al (2015) Curcumin improves amyloid β-peptide (1–42) induced spatial memory deficits through BDNF-ERK signaling pathway. PLoS ONE 10:1–17. https://doi.org/10.1371/journal.pone.0131525
Acknowledgements
I would like to thank SPPSPTM, SVKM’s NMIMS University, Mumbai, for their support.
Author information
Authors and Affiliations
Contributions
Conceptualization, GK and BT; methodology, GK and BT; software, BT; investigation, BT; writing—original draft preparation, BT; writing, review, and editing, GK; supervision, GK; All the authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Ethics approval
The Institutional Animal Ethics Committee permitted the study protocol (approval no. CPCSEA/IAEC/P-18/2019 dated March 22, 2019), which was formed following the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Government of India, and The National Institutes of Health (NIH) guidelines for the care of experimental animals were followed.
Consent to participate
NA.
Consent for publication
NA.
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Highlights
• Betanin ameliorates cognitive dysfunction in a scopolamine-induced zebrafish model.
• In a novel tank test, betanin improves exploratory behavior.
• Betanin promotes zebrafish novelty response and spatial memory in the Y maze test.
• Betanin significantly decreases AChE-specific activity.
• In zebrafish, betanin therapy dramatically increases the antioxidant state of the brain.
• Betanin treatment raises BDNF levels in the brain of zebrafish.
• Betanin (50 and 100 mg/L) showed promising cognitive-enhancing potential.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Thawkar, B.S., Kaur, G. Betanin mitigates scopolamine-induced cognitive impairment by restoring cholinergic function, boosting brain antioxidative status, and increasing BDNF level in the zebrafish model. Fish Physiol Biochem 49, 335–349 (2023). https://doi.org/10.1007/s10695-023-01185-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10695-023-01185-6