Active caspase-3 detection to evaluate apoptosis induced by Verbena officinalis essential oil and citral in chronic lymphocytic leukaemia cells

Abstract

Verbena officinalis L., Verbenaceae, commonly known as vervain, is a plant widely used in medicine. Despite of its widespread use in different traditional practices, the mechanisms of pharmacological actions of the plant and its volatile oil are still unclear. We evaluated the pro-apoptotic activity of V. officinalis essential oil and of its main component, citral, on lymphocytes collected from ten patients with chronic lymphocytic leukaemia (CLL), a disease in which a faulty apoptotic mechanism is still retained one of the primary pathogenic events, by adding to treated mononuclear cells, annexin-V, propidium iodide, and CD19. Apoptosis was also evaluated using anti-active-caspase-3 monoclonal antibody after permeabilization of the cells. Both V. officinalis essential oil and citral were found able to induce apoptosis in CLL cells and to activate caspase-3, which is considered the way by means they active apoptosis in B neoplastic cells. This data further support evidences that indicate natural compounds as possible lead structure to develop new therapeutic agents for CLL.

Differential Cytotoxic Activity of Essential Oil of Lippia citriodora from Different Regions in Morocco

“…Furthermore, among 72 peaks revealed by chromatography, 64 peaks (96%) were identified (according to the National Institute of Standards and Technology database). 24 Peaks represent about 80% of the compounds of extracted essential oils ( Table 1).…”

“…The extrinsic pathway is mediated by death receptors, whereas the intrinsic one (mitochondrial pathway) is triggered by the release of apoptogenic proteins such as cytochrome c, which activate caspase proteins that are the main effector molecules that induce this process. [23] In a previous study, De Martino et al [24] have reported that caspase-3 is an active cell-death protease involved in the execution phase of apoptosis, where cells undergo morphological changes such as DNA fragmentation, chromatin condensation and apoptotic body formation. In our study, we think that caspase-3 might be involved in the apoptotic effect of essential oil.…”

Verbena officinalis (Common Vervain) – A Review on the Investigations of This Medicinally Important Plant Species

“…The V. officinalis herb is also characterized by the presence of an essential oil, which consists of about 40 compounds, mainly, monoterpenoids (citral, limonene, cineole, carvone) [42] (▶ Table 2). The composition of V. officinalis essential oil depends on environmental conditions, V. officinalis chemotype tested, type of plant material, and the essential oil extraction method used.…”

“…The composition of V. officinalis essential oil depends on environmental conditions, V. officinalis chemotype tested, type of plant material, and the essential oil extraction method used. Hydrodistilled essential oil obtained from fresh plants grown in Italy revealed substantial amounts of citral (> 45 %) and isobornyl formate (> 40 %) [30,[42][43][44][45]. Steam-distilled essential oil from dried Moroccan plants contained mostly spathulenol (> 10 %), limonene, and eucalyptol (7.5 % each) [46].…”

“…In 2012, at Forman Christian College in Pakistan, research was conducted on the antibacterial activity of ethanolic extracts from the stems, leaves, and roots of V. officinalis using 13 bacterial strains that cause serious infections (including four methicillin-re- [46,47] α-curcumane 6.8 [47] β-caryophyllene 0.1-1.9 [30,42,44,46,47,70] trans-nerolidol 1.5 [47] bicyclosesquiphellandrene 1.3 [47] δ-cadinene 0.7-1.1 [46,47] β-bourbonene 0.6-1.1 [46,47] allo-aromadendrene 0.1-0.6 [30,42,44,47,70] α-cubenene 0.6 [47] γ-cadinene 0.5 [47] continued sistant Staphylococcus aureus strains and four multidrug-resistant Salmonella typhi strains). All the tested extracts showed a clear antibacterial effect on the strains used.…”