Evaluation of seeds ethanolic extracts of Triplaris gardneriana Wedd. using in vitro and in vivo toxicological methods

ABSTRACT

Triplaris gardneriana Wedd. is a tree used in folk medicine to treat venereal diseases and inflammation as well as a source of biological compounds with antioxidant capacity. In order to assess the safety of these bioactive compounds, the present study aimed to determine the toxicity of an ethanolic extract of T. gardneriana, (EETg). Toxicological tests included hemolytic activity, toxicity toward the brine shrimp Artemia, cytotoxicity against breast cancer cells (MCF7) and acute oral toxicity in rodents. In addition, toxicogenomics techniques were used to determine genome expression in MCF7 cells exposed to EETg. The results showed that the extract exhibits approximately 60% of hemolytic activity at the highest tested concentration (64 µg/ml) and toxicity against nauplii of Artemia sp. (LC₅₀ of 67.85 µg/ml). Further, EETg appears to be cytotoxic to MCF7 (cell viability reduced to 40% at 250 µg/ml after 24 hr). Genomic data demonstrated differential expression of 14 genes. Data analysis indicated possible altered pathways (e.g., xenobiotic metabolism), possible adverse health risks (e.g., hepatotoxicity), and drugs with similar gene expression profile (e.g., antimicrobials). The investigation provides important information on potentially adverse aspects of EETg, which need to be considered prior to the therapeutic utilization of this plant.

Abbreviations: EETg: ethanolic extract of T. gardneriana seeds; MCF7: michigan cancer foundation-7 which refers to a human breast cell line (adenocarcinoma); NGS: next-generation sequencing; edgeR: empirical analysis of digital gene expression data in R; Consensus: consensus path database; FDR: false discovery rate; NCBI: national center for biotechnology information; KEGG: kyoto encyclopedia of genes and genomes; Ingenuity: ingenuity pathway analysis software; CMAP: connectivity map; OECD: organization for economic co-operation and development; HL-60: human promyelocytic leukemia cells; PC3: prostate cancer cells.

KEYWORDS:

• Medicinal plant
• ATP-Lite
• toxicogenomics
• toxicity
• MCF7 cells
• quercetin

Conflict of interests

The authors declare that there is no conflict of interest regarding the publication of this manuscript.

Ethics approval

All animals used and the biological material were discarded according to the Biosafety and Bioethics Norms and Procedures (BAZZANO, 2006). The protocol adopted in this work was previously approved by the UFC Committee of Ethics in Animal Research (CEUA, No.105-17). The assay using human red blood cells was performed after being approved by the Ethics and Research Committee (CEP) of the Federal University of Ceará (registration number 3.277.799).

Submission declaration and verification

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

Authorship contributions

Thiago S. Almeida, José J. Lopes Neto, Igor P. Pessoa, Jackeline L. Medeiros, Pedro M. S. Tabosa, Thais B. Moreira, performed experiments and analyzed results. Terezinha M. Souza, Mariana R. Arantes carried out the statistical analysis. Thiago S. Almeida, Davi F. Farias, Ana F. U. Carvalho wrote the report and designed the research.

Additional information

Funding

This investigation in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001, for supporting grants (PVE-A006_2013).