Molecules (Jul 2023)

Cytotoxicity and Genotoxicity Evaluation of <i>Zanthoxylum rhoifolium</i> Lam and In Silico Studies of Its Alkaloids

  • Rufine Azonsivo,
  • Kelly Cristina Oliveira de Albuquerque,
  • Ana Laura Gadelha Castro,
  • Juliana Correa-Barbosa,
  • Helena Joseane Raiol de Souza,
  • Andryo Orfi de Almada-Vilhena,
  • Gleison Gonçalves Ferreira,
  • Anderson Albuquerque de Souza,
  • Andrey Moacir do Rosario Marinho,
  • Sandro Percario,
  • Cleusa Yoshiko Nagamachi,
  • Julio Cesar Pieczarka,
  • Maria Fâni Dolabela

DOI
https://doi.org/10.3390/molecules28145336
Journal volume & issue
Vol. 28, no. 14
p. 5336

Abstract

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The alkaloids isolated from Zanthoxylum rhoifolium have demonstrated great pharmacological potential; however, the toxic profiles of these extracts and fractions are still not well elucidated. This study evaluated the toxicity of the ethanol extract (EEZR) and neutral (FNZR) and alkaloid (FAZR) fractions. Chemical characterization was performed by chromatographic methods: thin-layer chromatography (TLC) and high-performance liquid chromatography coupled with diode array detection (HPLC–DAD). The cytotoxicity of the samples was evaluated in human hepatocellular carcinoma (HepG2) cells using the cell viability method (MTT) and mutagenicity by the Allium cepa assay (ACA). Alkaloids isolated from the species were selected for toxicity prediction using preADMET and PROTOX. The molecular docking of the topoisomerase II protein (TOPOII) was used to investigate the mechanism of cell damage. In the EEZR, FNZR, and FAZR, the presence of alkaloids was detected in TCL and HPLC–DAD analyses. These samples showed a 50% inhibitory concentration (IC50) greater than 400 μg/mL in HepG2 cells. In ACA, time- and concentration-dependent changes were observed, with a significant reduction in the mitotic index and an increase in chromosomal aberrations for all samples. Nuclear sprouts and a micronucleus of the positive control (PC) were observed at 10 µg/mL and in the FAZR at 30 µg/mL; a chromosomal bridge in FNZR was observed at 105 µg/mL, CP at a concentration of 40 µg/mL, and nuclear bud and mitotic abnormalities in the EEZR were observed at 170 µg/mL. The alkaloids with a benzophenanthridine were selected for the in silico study, as structural alterations demonstrated certain toxic effects. Molecular docking with topo II demonstrated that all alkaloids bind to the protein. In summary, the fractionation of Z. rhoifolium did not interfere with toxicity; it seems that alkaloids with a benzophenanthridine nucleus may be involved in this toxicity.

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