Phytochemical composition, cytotoxicity and acute toxicity of the ethyl acetate extract from Clidemia hirta leaves

Phytochemical composition, cytotoxicity and acute toxicity of the ethyl acetate extract from Clidemia hirta leaves

• Author
• STELLA DE JESUS LOURENCO DA SILVA
• Co-authors
• Elizabeth Fernanda de Oliveira Borba , Tonny Cley Campos Leite , Marília Grasielly de Farias Silva , Abner Lins Dantas , Nara de Melo Santos , Marley Gabriel da Silva Santos , Teresinha Gonçalves da Silva
• Abstract

• Phytochemical composition, cytotoxicity, and acute toxicity of the ethyl acetate extract from Clidemia hirta leaves

   

  Stella de Jesus Lourenço da Silva ¹; Elizabeth Fernanda de Oliveira Borba ²; Tonny Cley Campos Leite ³; Marília Grasielly de Farias Silva ⁴; Abner Lins Dantas ⁵; Nara de Melo Santos ⁶;  Marley Gabriel da Silva Santos ⁷; Teresinha Gonçalves da Silva ⁸

  ^{1, 2, 4, 5, 6,7,8} Federal University of Pernambuco (UFPE), Recife, Pernambuco, Brazil.

  ³ Federal Institute of Education, Science and Technology of Pernambuco (IFPE), Barreiros, Pernambuco, Brazil.

   

  stella.lourenco99@gmail.com; teresinha.goncalves@ufpe.br

   

  ABSTRACT

  INTRODUCTION: Cancer remains a challenging disease worldwide. The treatments available frequently present side effects and toxicity [1]. Given this, medicinal plants provide a great resource for discovering new anticancer biomolecules [2]. Clidemia hirta, known as ‘caiua’, is commonly used to treat wounds, gastrointestinal disorders, and tumors [3]. These uses are supported by the presence of tannins, flavonoids, and phenolic compounds, which have been associated with antimicrobial, antioxidant and anticancer activities [4,5]. AIMS: This work aimed to evaluate the phytochemical components, cytotoxicity, and acute toxicity of the ethyl acetate extract from C. hirta leaves (ChAcOEt). METHODS: The species was collected in Barreiros (PE-Brazil), and then identified at the Herbarium of the Agronomic Research Institute. The crude extract was obtained by exhaustive maceration of the dried leaves in ethyl acetate, and the solvent was removed in a rotatory evaporator. The phytochemistry was evaluated by the thin layer chromatography, and the determination of flavonoids, total phenolics, and tannins by the Aluminium Chloride (AlCl₃) and Folin-Ciocalteu methods, respectively. The cytotoxicity of ChAcOEt on non-tumor (L-929, VERO) and tumor (HT-29, NCI-H292, MCF-7) lineages was performed using the MTT (3-(4,5, dimethylthiazol-2-yl)-2,5 ditetrazolium bromide) assay [6]. Finally, the acute toxicity was analyzed according to the guide no. 423 from the Organization for Economic Co-operation and Development (OECD, 2001). The animal experimentation was approved by the ethical committee of the Federal University of Pernambuco (issue nº 0058/2022). RESULTS AND DISCUSSION: The phytochemical profile showed the presence of flavonoids and tannins. The total phenolics, flavonoids, and tannins resulted in 388. 6 mg EQ.A.G./g; 441.2 mg EQ.Rut./g, and 223,1 mg EQ.A.T./g, respectively. ChAcOEt presented percentages of growth inhibition above 30 % for L929 and VERO. These results allow future pharmacological studies of ChAcOEt, as it did not show significant cytotoxicity on healthy cells [7]. Furthermore, the extract inhibited more than 70 % of cell growth against HT-29, NCI-H292, and MCF-7, showing inhibition of 50% of the cells at the respective concentrations (IC₅₀): 19.6 ?g/mL, 6.4 ?g/mL, and 68.2 ?g/mL. Plant extracts can be considered cytotoxic against cancer cells when they exhibit an IC₅₀ under 30 ?g/mL [8]. The acute toxicity tests revealed no physiological, haematological, or biochemical changes in mice (2000 mg/kg) compared to the control. CONCLUSION: The ChAcOEt showed tannins and flavonoids in its composition, and cytotoxicity against the cancer lineages tested. Besides that, ChAcOEt did not exhibit toxicity in healthy cells or mice. However, it is crucial to identify the molecules responsible for this activity and their mechanism of action. ACKNOWLEDGEMENT: Thanks to the National Council for Scientific and Technological Development for providing financial support.

   

  Keywords: Cancer, Phytochemistry, Medicinal plants, Toxicology.

   

  REFERENCES

  [1] ASHRAFIZAVEH, S. et al. Long non-coding RNAs in the doxorubicin resistance of cancer cells. Cancer Letters, v. 508, p. 104-114, 2021.

  [2] SÜNTAR, Ipek. Importance of ethnopharmacological studies in drug discovery: role of medicinal plants. Phytochemistry Reviews, v. 19, n. 5, p. 1199-1209, 2020.

  [3] MEDEIROS, J. D. Guia de campo: vegetação do Cerrado 500 espécies. 1. ed. Brasília: MMA/SBF, 2011.

  [4] NARASIMHAM, D. et al. Evaluation of in vitro anticancer and antioxidant activities from leaf extracts of medicinal plant Clidemia hirta. International Journal of Pharmaceuctical Science, [S.I], v. 9, n. 4, p. 149-153, 2017.

  [5] BOMFIM, E. M. S. et al. Phytochemical composition and biological activities of extracts from ten species of the family Melastomataceae Juss. Brazilian Journal of Biology, São Paulo, v. 82, , e242112, 2021.

  [6] MOSMANN, T. Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and Cytotoxicity Assays. Journal of Immunological Methods, [Amsterdã], v. 65, n. 1-2, p. 55 – 63, 1983.

  [7] ARISTIZABAL, L. S. R.; MARÍN, D.; GONZÁLEZ, F. J. J.  Atividade ictiotóxica e citotóxica de extratos vegetais Chrysobalanaceae, Melastomataceae, Rubiaceae e Rutaceae, da flora colombiana. Scientia et Technica Año XVIII, [S.I], vol. 18, n. 3, p. 548- 552, 2013.

   

  [8] HENNEBELLE, T. et al. Ethnopharmacology of Lippia alba. Journal of ethnopharmacology, [Ireland], v. 116, n. 2, p. 211-222, 2008.

• Keywords
• Cancer, Phytochemistry, Medicinal plants, Toxicology
• Modality
• Pôster
• Subject Area
• Drug Design and Discovery, Synthesis and Natural Products