Antimitotic and toxicogenetic action of Stevia urticifolia aerial parts on proliferating vegetal and mammalian cells: in vitro and in vivo traditional and replacement methods

ABSTRACT

Stevia urticifolia Thunb. is an underexploited herb possessing bioactive flavonoids, saponins, and terpenoids. The aim of this study was to examine the antiproliferative and toxicogenetic properties of the ethyl acetate extract from Stevia urticifolia aerial parts (EtAcSur) upon Artemia salina, erythrocytes, Allium cepa and sarcoma 180 cells and fibroblasts, as well as in vivo studies on mice to determine systemic, macroscopic, and behavioral alterations and bone marrow chromosomal damage. The assessment using A. salina larvae and mouse blood cells revealed LC₅₀ and EC₅₀ values of 68.9 and 113.6 µg/ml, respectively. Root growth and mitosis were inhibited by EtAcSur, and chromosomal aberrations were detected only at 100 μg/ml. EtAcSur exhibited potent concentration-dependent viability reduction of S180 and L-929 cells and antioxidant capacity employing ABTS^• and DPPH^•. No previous in vivo studies were performed before with the EtAcSur. Signals of acute toxicity were not observed at 300 mg/kg. Physiological and toxicological investigations at 25 and 50 mg/mg/day i.p. for 8 days did not markedly change body or organ relative weights, nor patterns of spontaneous locomotor and exploratory activities. In contrast, clastogenic effects on bone marrow were found at 50 mg/mg/day. EtAcSur was found to (1) produce toxicity in microcrustaceans, (2) capacity as free radical scavenger, (3) antimitotic, cytotoxic and clastogenic activties upon vegetal and mammalian cells, and (4) lethality on both tumor and normal murine cells indistinctly. In vivo damage systemic effects were not remarkable and clinical signals of toxicity were not observed, suggesting the significant pharmacological potential of S. urticifolia for the development of antineoplastic agents.

Abbreviations: ABTS: 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid); DMSO: dimethylsulfoxide; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EC₅₀: effective concentration 50%; EtAcSur: ethyl acetate extract from Stevia urticifolia aerial parts; Hb, hemoglobin; IC₅₀: inhibitory concentration 50%; LC₅₀,: lethal concentration 50%; MI: mitotic index; RBC, red blood cells; Trolox: 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid.

KEYWORDS:

• Allium cepa
• chemoprevention
• in vivo toxicity
• genotoxicity
• sarcoma 180

Acknowledgments

Paulo Michel Pinheiro Ferreira and José Roberto de Oliveira Ferreira are grateful to the public Brazilian agencies “Conselho Nacional de Desenvolvimento Científico e Tecnológico” [CNPq (#303247/2019-3)] and “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior” [CAPES (Finance code 001)] for their personal scholarship, respectively. We also thank Leane Brunelle dos Santos Alves for technical assistance and the Postgraduate Program in Pharmaceutical Sciences (PPGCF, Teresina, Piauí, Brazil) for structural support.

Data availability statement

Raw data were generated at GraphPad Prisma. Derived data supporting the findings of this study are available from the corresponding author [P.M.P.F.] on request.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Ethics approval

The registry in SisGen (Sistema Nacional de Gestão do Patrimônio Genético e do Conhecimento Tradicional Associado – National System of Management of Genetic Heritage and Associated Traditional Knowledge: #AF467FD) according to the Brazilian Federal Law No 13,123/2015 about access to the national biodiversity (Brazil 2015). All protocols involving animals were approved by the Ethical Committee on Animal Experimentation at UFPI (CEUA #008/2015 and #0555/2019) and followed Brazilian (Sociedade Brasileira de Ciência em Animais de Laboratório - SBCAL) and International (Directive 2010/63/EU of the European Parliament and of the Council on the protection of animals used for scientific purposes) rules on the care and use of experimental animals.

Submission declaration and verification

The work described has not been previously published or submitted for publication elsewhere. The publication is approved by all authors.

Additional information

Funding

This investigation was partially supported by the Brazilian agency “Conselho Nacional de Desenvolvimento Científico e Tecnológico” [CNPq (#427366/2018-6)].