Skip to main content

Advertisement

SpringerLink
Log in

Early Detection of Toxic Cyanobacteria in Bulgarian Dam Water and In Vitro Evaluation of the Effect of Saponins From Astragalus glycyphyllos and A. glycyphylloides, in Cyanotoxin (Anatoxin-α)-Induced Neurotoxicity

  • Original Article
  • Published:
Revista Brasileira de Farmacognosia Aims and scope Submit manuscript

Abstract

As a result of the blooming, certain Cyanobacteria (i.e., Microcystis) produce toxins, at concentrations that could be toxic. In this study, a group of the early detection markers for toxic microalgae were selected and applied. Samples from three Bulgarian dams, used as a source of drinking water for the general population, were tested via real time PCR. To monitor the presence of bacterial DNA, specific primers were selected to demonstrate Cyanobactеria. Some Cyanobacteria produced anatoxin-α, which is neurotoxic. The possible neuroprotective effect of two saponins, isolated from Astragalus glycyphyllos L. and A. glycyphylloides DC., Fabaceae, in a model of cyanotoxin (anatoxin-α)-induced neurotoxicity on subcellular fractions (rat brain microsomes, mitochondria, synaptosomes) and at cellular (neuroblastoma cell line SH-SY5Y) level, were further investigated. For preliminary assessment of physicochemical properties, pharmacokinetics, and drug-likeness of the investigated compounds, an in silico approach was applied. Anatoxin-α did not exert neurotoxic effect in SH-SY5Y cell line. The data results showed that the saponin from A. glycyphyllos had more pronounced neuroprotective effect on subcellular fractions, compared with the saponin from A. glycyphylloides. These results were supported by the in silico data analysis, where A. glycyphyllos saponin had better physicochemical and pharmacokinetics profile than the saponin from A. glycyphylloides.

Graphical abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Antoine D, Olivier M, Vincent Z (2017) SwissADME: a free web tool to evaluate pharmacokinetics, druglikeness and medicinal chemistry friendliness of small molecules. Sci Rep 7:42717. https://doi.org/10.1038/srep42717

    Article  Google Scholar 

  • Aráoz R, Molgó J, de Marsac NT (2010) Neurotoxic cyanobacterial toxins. Toxicon 56:813–828

    Article  Google Scholar 

  • Auyeung KK, Han Q-B, Ko J (2016) Astragalus membranaceus: a review of its protection against inflammation and gastrointestinal cancers. Am J Chin Med 44:1–22. https://doi.org/10.1142/S0192415X16500014

    Article  PubMed  Google Scholar 

  • Chan W-S, Durairajan SSK, Lu J-H, Wang Y, Xie L-X, Kum W-F, Koo I, Yung KKL, Li M (2009) Neuroprotective effects of Astragaloside IV in 6-hydroxydopamine-treated primary nigral cell culture. Neurochem Int 55:414–422

    Article  CAS  Google Scholar 

  • Dahlin J, Inglese J, Walters M (2015) Mitigating risk in academic preclinical drug discovery. Nature Rev Drug Discov 14:279–294. https://doi.org/10.1038/nrd4578

    Article  CAS  Google Scholar 

  • Forster JI, Köglsberger S, Trefois C, Boyd O, Baumuratov AS, Buck L, Balling R, Antony PMA (2016) Characterization of differentiated SH-SY5Y as neuronal screening model reveals increased oxidative vulnerability. J Biomol Screen 21:496–509. https://doi.org/10.1177/1087057115625190

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hawkins B, Bosch S, Odin M, Wohlers D, Donohue J, Hilborn E,Sinclair J (2006) Toxicological review of cyanobacterial toxins: anatoxin-a (external review draft). U.S. Environmental Protection Agency, Washington, DC, EPA/600/R-06/137

  • Hughes JP, Rees S, Kalindjian SB, Philpott KL (2011) Principles of early drug discovery. Br J Pharmacol 162:1239–1249

    Article  CAS  Google Scholar 

  • Kellmann R, Ploux O, Neilan BA (2013) Neurotoxic alkaloids from Cyanobacteria. In: Ramawat KG, Mérillon J-M (eds) Natural products: Phytochemistry, botany and metabolism of alkaloids. Phenolics and Terpenes. Springer, Berlin, pp 39–83. https://doi.org/10.1007/978-3-642-22144-6_47

    Chapter  Google Scholar 

  • Kondeva-Burdina M, Voynova M, Shkondrov A, Aluani D, Tzankova V, Krasteva I (2019) Effects of Amanita muscaria extract on different in vitro neurotoxicity models at sub-cellular and cellular levels. Food Chem Toxicol. https://doi.org/10.1016/j.fct.2019.110687

  • Krasteva I, Shkondrov A, Ionkova I, Zdraveva P (2016) Advances in phytochemistry, pharmacology and biotechnology of Bulgarian Astragalus species. Phytochem Rev 15:567–590. https://doi.org/10.1007/s11101-016-9462-4

    Article  CAS  Google Scholar 

  • Kurmayer R, Kutzenberger T (2003) Application of real-time PCR for quantification of microcystin genotypes in a population of the toxic Cyanobacterium microcystis sp. Appl Environ Microbiol 69:6723–6730

    Article  CAS  Google Scholar 

  • Lee D-Y, Noh H-J, Choi J, Lee K-H, Lee M-H, Lee J-H, Hong Y, Lee S-E, Kim S-Y, Kim G-S (2013) Anti-inflammatory cycloartane-type saponins of Astragalus membranaceus. Molecules 18:3725–3732

    Article  CAS  Google Scholar 

  • Lysiuk R, Darmohray R (2016) Pharmacology and ethnomedicine of the genus Astragalus. Int J Pharmacol Phytochem Ethnomed 3:46–53

    Google Scholar 

  • Mansuy D, Sassi A, Dansette PM, Plat M (1986) A new potent inhibitor of lipid peroxidation in vitro and in vivo, the hepatoprotective drug anisyldithiolthione. Biochem Biophys Res Commun 135:1015–1021. https://doi.org/10.1016/0006-291X(86)91029-6

    Article  CAS  PubMed  Google Scholar 

  • Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63. https://doi.org/10.1016/0022-1759(83)90303-4

    Article  CAS  Google Scholar 

  • Mungarro-Menchaca X, Ferrera P, Morán J, Arias C (2002) β-Amyloid peptide induces ultrastructural changes in synaptosomes and potentiates mitochondrial dysfunction in the presence of ryanodine. J Neurosci Res 68:89–96

    Article  CAS  Google Scholar 

  • Pavlova V, Babica P, Todorova D, Bratanova Z, Maršálek B (2006) Contamination of some reservoirs and lakes in Republic of Bulgaria by microcystins. Acta Hydrochim Hydrobiol 34:437–441

    Article  CAS  Google Scholar 

  • Rao PL, Bhattacharya R, Gupta N, Parida M, Bhaskar A, Dubey R (2002) Involvement of caspase and reactive oxygen species in cyanobacterial toxin anatoxin-a-induced cytotoxicity and apoptosis in rat thymocytes and Vero cells. Arch Toxicol 76:227–235

    Article  Google Scholar 

  • Robyt JF, Ackerman RJ, Chittenden CG (1971) Reaction of protein disulfide groups with Ellman’s reagent: a case study of the number of sulfhydryl and disulfide groups in Aspergillus oryzae α-amylase, papain, and lysozyme. Arch Biochem Biophys 147:262–269

    Article  CAS  Google Scholar 

  • Rodgers KJ, Main BJ, Samardzic K (2018) Cyanobacterial neurotoxins: their occurrence and mechanisms of toxicity. Neurotox Res 33:168–177. https://doi.org/10.1007/s12640-017-9757-2

    Article  CAS  PubMed  Google Scholar 

  • Rymuszka A (2012) Cytotoxic activity of the neurotoxin anatoxin-a on fish leukocytes in vitro and in vivo studies. Acta Vet Brno 81:175–182

    Article  CAS  Google Scholar 

  • Shirani M, Alizadeh S, Mahdavinia M, Dehghani MA (2019) The ameliorative effect of quercetin on bisphenol A-induced toxicity in mitochondria isolated from rats. Environ Sci Pollut Res 26:7688–7696. https://doi.org/10.1007/s11356-018-04119-5

    Article  CAS  Google Scholar 

  • Shkondrov A, Krasteva I, Bucar F, Kunert O, Kondeva-Burdina M, Ionkova I (2018) Flavonoids and saponins from two Bulgarian Astragalus species and their neuroprotective activity. Phytochem Lett 26:44–49. https://doi.org/10.1016/j.phytol.2018.05.015

    Article  CAS  Google Scholar 

  • Shkondrov A, Krasteva I, Bucar F, Kunert O, Kondeva-Burdina M, Ionkova I (2020) A new tetracyclic saponin from Astragalus glycyphyllos L. and its neuroprotective and hMAO-B inhibiting activity. Nat Prod Res 34:511–517. https://doi.org/10.1080/14786419.2018.1491040

    Article  CAS  PubMed  Google Scholar 

  • Sims NR, Anderson MF (2008) Isolation of mitochondria from rat brain using Percoll density gradient centrifugation. Nat Protoc 3:1228–1239

    Article  CAS  Google Scholar 

  • Stoyneva-Gärtner MP, Descy J-P, Latli A, Uzunov BA, Pavlova VT, Bratanova Z, Babica P, Maršálek B, Meriluoto J, Spoof L (2017) Assessment of cyanoprokaryote blooms and of cyanotoxins in Bulgaria in a 15-years period (2000-2015). Adv Oceanogr Limnol 8:131–152

    Article  Google Scholar 

  • Takser L, Benachour N, Husk B, Cabana H, Gris D (2016) Cyanotoxins at low doses induce apoptosis and inflammatory effects in murine brain cells: potential implications for neurodegenerative diseases. Toxicol Reports 3:180–189

    Article  CAS  Google Scholar 

  • Taupin P, Zini S, Cesselin F, Ben-Ari Y, Roisin M-P (1994) Subcellular fractionation on Percoll gradient of mossy fiber synaptosomes: morphological and biochemical characterization in control and degranulated rat hippocampus. J Neurochem 62:1586–1595

    Article  CAS  Google Scholar 

  • Tillett D, Neilan BA (2000) Xanthogenate nucleic acid isolation from cultured and environmental cyanobacteria. J Phycol 36:251–258

    Article  CAS  Google Scholar 

  • Xicoy H, Wieringa B, Martens GJM (2017) The SH-SY5Y cell line in Parkinson’s disease research: a systematic review. Mol Neurodegener. https://doi.org/10.1186/s13024-017-0149-0

Download references

Acknowledgements

Authors are grateful for the technical support from the whole team of “Genetically Modified Organisms” Department, National Center of Public Health and Analyses, Sofia, Bulgaria.

Funding

The study was carried out with the financial support of the Council of Medicinal Science at Medical University of Sofia, Contract no. D-145/2019, project no. 8321/22.11.2018.

Author information

Authors and Affiliations

  1. Laboratory of Drug Metabolism and Drug Toxicity, Section of Toxicology, Department of Pharmacology, Pharmacotherapy and Toxicology, Faculty of Pharmacy, Medical University of Sofia, 2 Dunav St., 1000, Sofia, Bulgaria

    Victoria Ilieva, Denitsa Aluani, Magdalena Kondeva-Burdina & Virginia Tzankova

  2. Department of Pharmacognosy, Faculty of Pharmacy, Medical University of Sofia, Sofia, Bulgaria

    Aleksandar Shkondrov & Ilina Krasteva

  3. Department of Chemistry, Faculty of Pharmacy, Medical University of Sofia, Sofia, Bulgaria

    Iva Valkova

  4. Laboratory of Genetically Modified Organisms, National Center of Public Health and Analyses, Sofia, Bulgaria

    Tzveta Georgieva

  5. Laboratory of Applied Biotechnologies, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, Plovdiv, Bulgaria

    Milen Georgiev

Authors
  1. Victoria Ilieva
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Aleksandar Shkondrov
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Denitsa Aluani
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Iva Valkova
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Magdalena Kondeva-Burdina
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tzveta Georgieva
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Virginia Tzankova
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ilina Krasteva
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Milen Georgiev
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

VI and MKB isolated rat brain mitochondria, microsomes, and synaptosomes and worked with them. DA and VT worked with SH-SY5Y. IV made the in silico analysis. VI and TG performed PCR experiment. AS and IK isolated saponins from Astragalus. MG critically revised the material and made the concept for it.

Corresponding author

Correspondence to Victoria Ilieva.

Ethics declarations

Conflict of Interest

The authors declare that they have no conflict of interest.

Confidentiality of Data

The authors declare that no patient data appear in this article.

Right to Privacy and Informed Consent

The authors declare that no patient data appear in this article.

Electronic Supplementary Material

ESM 1

(PDF 296 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ilieva, V., Shkondrov, A., Aluani, D. et al. Early Detection of Toxic Cyanobacteria in Bulgarian Dam Water and In Vitro Evaluation of the Effect of Saponins From Astragalus glycyphyllos and A. glycyphylloides, in Cyanotoxin (Anatoxin-α)-Induced Neurotoxicity. Rev. Bras. Farmacogn. 30, 202–213 (2020). https://doi.org/10.1007/s43450-020-00050-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43450-020-00050-4

Keywords

Search

Navigation