Evidence of biological activity of Mentha species extracts on apoptotic and autophagic targets on murine RAW264.7 and human U937 monocytic cells

Pharm Biol. 2017 Dec;55(1):286-293. doi: 10.1080/13880209.2016.1235208.

Abstract

Context: Mints (Lamiaceae) are used as traditional remedies for the treatment of several diseases. Their extracts are recognized as anti-inflammatory compounds.

Objective: This study characterized the cytotoxic effects of Mentha spicata L. (MS), Mentha pulegium L. (MP) and Mentha rotundifolia (L). Huds (MR) on macrophage cells (RAW264.7; U937) and determined their impact on apoptosis and autophagy, which can play a role in controlling inflammation.

Materials and methods: The extracts were prepared in culture medium and tested from 25 to 400 μg/mL after 24-48 h of treatment. To show the effect of the aqueous ethanol (50%) extracts on apoptosis and authophagy, the presence of cleaved caspase-3, and the conversion of LC3-I to LC3-II was evaluated by Western blotting.

Results: Compared with the MTT assay, crystal violet showed a pronounced decrease in the number of cells with all extracts at 48 h. Calculated IC50 values were 257.31, 207.82 and 368.02 μg/mL for MS, MP and MR, respectively. A significant increase in PI positive cells was observed with all extracts at 200-400 μg/mL. Mitochondrial dysfunctions and nuclear morphological changes were detected with MS and MR extracts at 400 μg/mL. At this concentration, no cleaved caspase-3 was found whereas stabilized caspase-3 in its dimeric form was identified. MS and MR extracts also favour LC3-I to LC3-II conversion which is a criterion of autophagy.

Conclusions: The cytotoxic profiles depend on the extracts considered; MS extract showed the strong activity. However, all the mint extracts studied interact with the apoptotic and autophagic pathways at elevated concentrations.

Keywords: Mentha spicata; mentha pulegium; mentha rotundifolia; mitochondria.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / isolation & purification
  • Anti-Inflammatory Agents / pharmacology*
  • Apoptosis / drug effects*
  • Autophagy / drug effects*
  • Caspase 3 / metabolism
  • Cell Survival / drug effects
  • Dose-Response Relationship, Drug
  • Humans
  • Inhibitory Concentration 50
  • Macrophages / drug effects*
  • Macrophages / metabolism
  • Macrophages / pathology
  • Membrane Potential, Mitochondrial / drug effects
  • Mentha / chemistry*
  • Mice
  • Microtubule-Associated Proteins / metabolism
  • Monocytes / drug effects*
  • Monocytes / metabolism
  • Monocytes / pathology
  • Phytotherapy
  • Plant Extracts / isolation & purification
  • Plant Extracts / pharmacology*
  • Plants, Medicinal
  • RAW 264.7 Cells
  • Time Factors
  • U937 Cells

Substances

  • Anti-Inflammatory Agents
  • Map1lc3b protein, mouse
  • Microtubule-Associated Proteins
  • Plant Extracts
  • Casp3 protein, mouse
  • Caspase 3