Abstract
Chlorella is a green alga consumed as dietary food supplement in pulverized form. In addition to its high nutritional value, it is reported as an excellent detoxifying agent. The pulverized Chlorella is partially soluble in water and insoluble portion has been reported for removal of mercury, cadmium and radioactive strontium from body. Chlorella contains a variety of metal-binding functional groups such as carboxyl, amino, phosphoryl, hydroxyl and carbonyl groups, which has high affinity towards various metal ions. The present study was envisaged to evaluate the chelating effect of water soluble fraction of Chlorella powder (AqCH) on metal ions. Fura-2 fluorescence ratio (F340/F380) was measured by fluorescence spectrometer (FS) after the exposure of chloride salt of metals viz., strontium, cobalt, barium, cesium, thallium and mercury to lymphocytes. Pretreatment of AqCH (0.1–20 mg mL−1) was given to evaluate the attenuating effect on fura-2 fluorescence ratio induced by metal ions. The intracellular levels of these metal ions were analyzed by atomic absorption spectrophotometer (AAS) and fluorescence microscopy (FM). Pretreatment with AqCH significantly attenuated the metal induced fluorescence ratio in dose-dependent manner. The results of AAS and FM were found in coherence with fura-2 fluorescence ratio which emphasized that AqCH significantly prevented the metal ions internalization. The present study suggests AqCH chelates with these metal ions and prevents its interaction with cells thereby reducing the intracellular mobilization of Ca2+.
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References
Amini M, Younesi H, Bahramifar N (2013) Biosorption of U(VI) from aqueous solution by Chlorella vulgaris: equilibrium, kinetic and thermodynamic studies. J Environ Eng 139:410–421. https://doi.org/10.1061/(ASCE)EE.1943-7870.0000651
Atar D, Backx PH, Appel MM, Gao WD, Marban E (1995) Excitation-transcription coupling mediated by zinc influx through voltage-dependent calcium channels. J Biol Chem 270:2473–2477
ATSDR (2004) Toxicological profile for strontium. Agency for Toxic Substances and Disease Registry, April 2004. https://www.atsdr.cdc.gov/ToxProfiles/tp159.pdf. Accessed 29 Dec 2018
Avery SV (1995) Caesium accumulation by microorganisms: uptake mechanisms, cation competition, compartmentalization and toxicity. J Ind Microbiol 14(2):76–84. https://doi.org/10.1007/BF01569888
Barkleit A, Moll H, Bernhard G (2008) Interaction of uranium(VI) with lipopolysaccharide. Dalton Trans 0:2879–2886
Benters J, Flögel U, Schäfer T, Leibfritz D, Hechtenberg S, Beyersmann D (1997) Study of the interactions of cadmium and zinc ions with cellular calcium homoeostasis using 19F-NMR spectroscopy. Biochem J 15(Pt 3):793–799. https://doi.org/10.1042/bj3220793
Bouton CM, Frelin LP, Forde CE, Godwin HA, Pevsner J (2001) Synaptotagmin I is a molecular target for lead. J Neurochem 76(6):1724–1735
Ciapetti G, Roda P, Landi L, Facchini A, Pizzoferrato A (1992) In vitro methods to evaluate metal–cell interactions. Int J Artif Organs 15(1):62–66. https://doi.org/10.1177/039139889201500111
Dakovic M, Popovic Z, Giester G, Rajic-Linaric M (2008) Thiocyanate complexes of the group 12 metals with pyridine-2-carboxamide: synthesis and structural characterization. Polyhedron 27(1):210–222. https://doi.org/10.1016/j.poly.2007.09.014
Edris G, Alhamed Y, Alzahrani A (2014) Biosorption of cadmium and lead from aqueous solutions by Chlorella vulgaris biomass: equilibrium and kinetic study. Arab J Sci Eng 39:87–93. https://doi.org/10.1007/s13369-013-0820-x
EPA (2017) Radionuclide basics: strontium-90. Radiation Protection. United States Environmental Protection Agency (US EPA), August 8 2017. https://www.epa.gov/radiation/radionuclide-basics-strontium-90. Accessed 10 June 2019
Granchi D, Ciapetti G, Savarino L, Cavedagna D, Donati ME, Pizzoferrato A (1996) Assessment of metal extract toxicity on human lymphocytes cultured in vitro. J Biomed Mater Res 31(2):183–191. https://doi.org/10.1002/(SICI)1097-4636(199606)31:2%3C183::AID-JBM4%3E3.0.CO;2-J
Hinkle PM, Shanshala ED II, Nelson EJ (1992) Measurement of intracellular cadmium with fluorescent dyes. Further evidence for the role of calcium channels in cadmium uptake. J Biol Chem 267:25553–25559
Kalin M, Wheeler WN, Meinrath G (2005) The removal of uranium from mining waste water using algal/microbial biomass. J Environ Radioact 78:151–177. https://doi.org/10.1016/j.jenvrad.2004.05.002
Kern M, Wisniewski M, Cabell L, Audesirk G (2000) Inorganic lead and calcium interact positively in activation of calmodulin. NeuroToxicology 21(3):353–364
Liang S, Liu X, Chen F, Chen Z (2004) Current microalgal health food R&D activities in China. Put O. Ang Jr. (ed.) Asian Pacific phycology in the 21st century: prospects and challenges. Hydrobiologia 512:45–48
Limke TL, Otero-Montañez JK, Atchison WD (2003) Evidence for interactions between intracellular calcium stores during methylmercury-induced intracellular calcium dysregulation in rat cerebellar granule neurons. J Pharmacol Exp Ther 304(3):949–958
Louis KS, Siegel AC (2011) cell viability analysis using trypan blue: manual and automated methods. Methods Mol Biol 740:7–12. https://doi.org/10.1007/978-1-61779-108-6_2
Marchetti C (2013) Role of calcium channels in heavy metal toxicity. ISRN Toxicol 30:1–9. https://doi.org/10.1155/2013/184360
Marchi B, Burlando B, Panfoli I, Viarengo A (2000) Interference of heavy metal cations with fluorescent Ca2+ probes does not affect Ca2+ measurements in living cells. Cell Calcium 28:225–231. https://doi.org/10.1054/ceca.2000.0155
Marczenko Z, Balcerzak M (2000) Uranium. Separation, preconcentration and spectrophotometry in inorganic analysis. Anal Spectr 10:446–455. https://doi.org/10.1016/s0926-4345(00)80118-2
Marty MS, Atchison WD (1998) Elevations of intracellular Ca2+ as a probable contributor to decreased viability in cerebellar granule cells following acute exposure to methylmercury. Toxicol Appl Pharmacol 150:98–105
Nicotera P, Bellomo G, Orrenius S (1992) Calcium-mediated mechanisms in chemically induced cell death. Annu Rev Pharmacol Toxicol 32:449–470
Ogawa K, Fukuda T, Han J, Kitamura Y, Shiba K, Odani A (2016) Evaluation of Chlorella as a decorporation agent to enhance the elimination of radioactive strontium from body. PLoS ONE 11:e0148080. https://doi.org/10.1371/journal.pone.0148080
Ohkubo M, Miyamoto A, Shiraishi M (2016) Heavy metal chelator TPEN attenuates fura-2 fluorescence changes induced by cadmium, mercury and methylmercury. J Vet Med Sci 78(5):761–767. https://doi.org/10.1292/jvms.15-0620
Ouyang H, Vogel HJ (1998) Metal ion binding to calmodulin: NMR and fluorescence studies. Biometals 11(3):213–222
Pantami HA, Ahamad Bustamam MS, Lee SY, Ismail IS, Mohd Faudzi SM, Nakakuni M, Shaari K (2020) Comprehensive GCMS and LC–MS/MS metabolite profiling of Chlorella vulgaris. Mar Drugs 18:367. https://doi.org/10.3390/md18070367
Pantami HA, Shaari K, Ahamad Bustamam MS, Ismail IS (2020) Metabolite profiling of different solvent extracts of the microalgae Chlorella vulgaris via 1H NMR-based metabolomics. Curr Metab Syst Biol 7:1–11
Peppiatt CM, Collins TJ, Mackenzie L, Conway SJ, Holmes AB, Bootman MD, Berridge MJ, Seo JT, Roderick HL (2003) 2-Aminoethoxydiphenyl borate (2-APB) antagonises inositol 1,4,5-trisphosphate-induced calcium release, inhibits calcium pumps and has a use-dependent and slowly reversible action on store-operated calcium entry channels. Cell Calcium 34:97–108
Prananto YP, Urbatsch A, Moubaraki B, Murray KS, Turner DR, Deacon GB, Batten SR (2017) Transition metal thiocyanate complexes of picolylcyanoacetamides. Aust J Chem 70:516–528. https://doi.org/10.1071/CH16648
Rezaei H, Kulkarni SD, Saptarshi PG (2012) Study of physical chemistry on biosorption of zinc by using Chlorella pyrenoidosa. Russ J Phys Chem 86:1332–1339. https://doi.org/10.1134/S0036024412060118
Safi C, Zebib B, Merah O, Pontalier PY, Vaca-Garcia C (2014) Morphology, composition, production, processing and applications of Chlorella vulgaris: a review. Renew Sustain Energy Rev 35:265–278. https://doi.org/10.1016/j.rser.2014.04.007
Shen QH, Zhi TT, Cheng LH, Xu XH, Chen HL (2013) Hexavalent chromium detoxification by nonliving Chlorella vulgaris cultivated under tuned conditions. Chem Eng J 228:993–1002. https://doi.org/10.1016/j.cej.2013.05.074
Shim JE, Son YA, Park JM, Kim MK (2009) Effect of Chlorella intake on cadmium metabolism in rats. Nutr Res Pract 3:15–22. https://doi.org/10.4162/nrp.2009.3.1.15
Silva-Pereira LC, Cardoso PC, Leite DS, Bahia MO, Bastos WR, Smith MA, Burbano RR (2005) Cytotoxicity and genotoxicity of low doses of mercury chloride and methylmercury chloride on human lymphocytes in vitro. Braz J Med Biol Res 38(6):901–907. https://doi.org/10.1590/s0100-879x2005000600012
Solisio C, Al Arni S, Converti A (2017) Adsorption of inorganic mercury from aqueous solutions onto dry biomass of Chlorella vulgaris: kinetic and isotherm study. Environ Technol 3330:1–9. https://doi.org/10.1080/09593330.2017.1400114
Sun X, Tian X, Tomsig JL, Suszkiw JB (1999) Analysis of differential effects of Pb2+ on protein kinase C isozymes. Toxicol Appl Pharmacol 156(1):40–45
Švadlenková M, Lukavský J, Kvíderová J (2005) Radionuclides 137Cs and 60Co uptake by freshwater and marine microalgae Chlorella, Navicula, Phaeodactylum. In: Bréchignac F, Desmet G (eds) Equidosimetry—ecological standardization and equidosimetry for radioecology and environmental ecology. Springer, Dordrecht, pp 379–387. https://doi.org/10.1007/1-4020-3650-7_44
Tan XX, Tang C, Castoldi AF, Manzo L, Costa LG (1993) Effects of inorganic and organic mercury on intracellular calcium levels in rat T lymphocytes. J Toxicol Environ Health 38:159–170
Thevenod F (2010) Catch me if you can! Novel aspects of cadmium transport in mammalian cells. Biometals 23:857–875
Vijayan p, Kumar SV, Dhanaraj SA, Badami S, Suresh B (2002) In vitro cytotoxicity and anti-tumor properties of the total alkaloid fraction of unripe fruits of Solanum pseudocapsicum. Pharm Biol 40:456–460
Vogel M, Gunther A, Rossberg A, Li B, Bernhard G, Raff J (2010) Biosorption of U(VI) by the green algae Chlorella vulgaris in dependence of pH value and cell activity. Sci Total Environ 409:384–395. https://doi.org/10.1016/j.scitotenv.2010.10.011
Wehrheim B, Wettern M (1994) Biosorption of cadmium, copper and lead by isolated mother cell walls and whole cells of Chlorella fusca. Appl Microbiol Biotechnol 41:725–728. https://doi.org/10.1007/BF00167291
Yadav M, Rani K, Chauhan MK, Panwar A, Sandal N (2020) Evaluation of mercury adsorption and removal efficacy of pulverized Chlorella (C. vulgaris). J Appl Phycol 32:1253–1262. https://doi.org/10.1007/s10811-020-02052-0
Yen HW, Chen PW, Hsu CY, Lee L (2017) The use of autotrophic Chlorella vulgaris in chromium(VI) reduction under different reduction conditions. J Taiwan Inst Chem Eng 74:1–6. https://doi.org/10.1016/j.jtice.2016.08.017
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The authors would like to thanks Director, Institute of Nuclear Medicine and Allied Sciences (INMAS), DRDO, New Delhi for providing all the necessary facilities and requirement to complete this study.
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Yadav, M., Soni, R., Chauhan, M.K. et al. Cellular and physiological approaches to evaluate the chelating effect of Chlorella on metal ion stressed lymphocytes. Biometals 34, 351–363 (2021). https://doi.org/10.1007/s10534-021-00285-1
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DOI: https://doi.org/10.1007/s10534-021-00285-1