Cytotoxicity of Cymbopogon citratus (DC) Stapf fractions, essential oil, citral, and geraniol in human leukocytes and erythrocytes.

ETHNOPHARMACOLOGY RELEVANCE: Our recently published paper demonstrated that ethyl acetate fractions obtained from Cymbopogon citratus (DC.) Stapf (C. citratus) leaves, which are consumed as infusion in folk medicine due to their therapeutic properties, are rich in polyphenols and exhibit promising antioxidant activity by acting through different mechanisms in vitro. However, studies regarding the toxicity of these fractions are necessary to investigate their safe use in future biomedical applications. AIM OF THE STUDY: This study aimed to investigate the toxicity of ethyl acetate (obtained in acidic and basic conditions and after the essential oil removal from the leaves) and chloroform fractions, essential oil, and its pure constituents, citral and geraniol. MATERIALS AND METHODS: The toxicity of C. citratus samples was evaluated by using Artemia salina (A. salina) and human blood cells (leukocytes and erythrocytes). RESULTS: The A. salina lethality assay demonstrated that C. citratus fractions were moderately toxic with LC50 values ranging from 146.12 to 433.15 µg mL-1, whereas the essential oil and isolated compounds were highly toxic with LC50 lower than 100 µg mL-1. Leukocyte viability decreased after incubation in the presence of the fractions obtained after the essential oil removal from the plant leaves, as well as in the presence of essential oil, citral and geraniol. The same samples increased the osmotic fragility of erythrocytes, and field emission gun scanning electron microscopy (FESEM) analysis revealed significant changes in cell morphology. Interestingly, our results suggest that the previous removal of essential oil from plant leaves facilitated the extraction of cytotoxic compounds from C. citratus. CONCLUSIONS: It was demonstrated that C. citratus ethyl acetate and chloroform fractions, essential oil, as well citral and geraniol were considered toxic to A. salina, cytotoxic to human blood cells and showed to induce alterations in the erythrocyte membrane at higher concentrations. These fractions will be further investigated to identify the phytochemicals involved in the observed cytotoxic effects and explored using in vivo models.

Cymbopogon citratus (DC.) Stapf, citral and geraniol exhibit anticonvulsant and neuroprotective effects in pentylenetetrazole-induced seizures in zebrafish.

ETHNOPHARMACOLOGICAL RELEVANCE: Cymbopogon citratus (DC.) Stapf (C. citratus) is consumed as an infusion in folk medicine due to its pharmacological properties and action in the central nervous system. Epilepsy is a neurological disorder that affects millions of people. Since the currently available antiepileptic drugs often cause undesirable side effects, new alternative therapeutic strategies based on medicinal plants have been proposed. AIM OF THE STUDY: This study aimed to investigate the anticonvulsant and neuroprotective effects of C. citratus essential oil (EO) and hydroalcoholic extract (E1) from its leaves, as well as of its related compounds citral (CIT) and geraniol (GER) against the effects of pentylenetetrazole (PTZ) induced seizures in zebrafish (Danio rerio). MATERIALS AND METHODS: To evaluate the anticonvulsant properties of the samples, adult animals were pre-treated (by immersion) and subsequently exposed to PTZ solution. The involvement of GABAA receptors in the antiepileptic effects was investigated by the coadministration of flumazenil (FMZ), a known GABAA receptor antagonist. Oxidative stress markers malondialdehyde (MDA), glutathione (GSH), catalase (CAT) and nitric oxide (NO) were assessed in zebrafish brain homogenates after PTZ exposure. RESULTS: All samples increased the latency time for the first seizure, which was reduced when animals were pretreated with FMZ, suggesting the involvement of GABAA receptors in the observed properties. The association between CIT and GER at the lowest concentration studied showed a synergistic effect on the anticonvulsant activity. Decreases in MDA and NO levels and increases in GSH and CAT levels in the brain of treated animals suggested the neuroprotective effect of the compounds investigated. CONCLUSIONS: Our results proved that C. citratus EO, E1, CIT and GER have anticonvulsant effects in zebrafish and could be used as a promising adjuvant therapeutic strategy for epilepsy treatment. Furthermore, zebrafish demonstrated to be an alternative animal model of epilepsy to evaluate the anticonvulsant and neuroprotective effects of C. citratus.

Anxiolytic properties of Cymbopogon citratus (DC.) stapf extract, essential oil and its constituents in zebrafish (Danio rerio).

ETHNOPHARMACOLOGICAL RELEVANCE: Cymbopogon citratus (DC.) Stapf (Poaceae) leaves is often consumed as infusion in folk medicine due to its therapeutic properties. This plant is also rich in essential oil, which has several beneficial effects to the human health. It is known that medications commonly used to treat anxiety disorders cause undesirable side effects. Thus, it is important to evaluate the anxiolytic effects of natural products from plants, such as C. citratus, as an alternative therapy to treat these disorders. OBJECTIVE: The aim of this study was to investigate the anxiolytic properties of C. citratus essential oil (EO), hydroalcoholic extract (E1), citral (CIT), geraniol (GER) and the mixture of these terpenoids, as well as its possible mechanism of action by using zebrafish as an anxiety model. METHODS: Adult zebrafish were treated (by immersion) with C. citratus EO, E1, CIT and/or GER. The anxiolytic effects were analyzed by using the light-dark test. The mechanism involved in the anxiolytic effects was further investigated by the coadministration of flumazenil (FMZ), an antagonist of GABAA receptors. The total polyphenols (phenolic and flavonoid compounds) content of E1 was determined by using spectrophotometric assays. RESULTS: All analyzed samples showed a remarkable anxiolytic effect on zebrafish in the highest concentrations, as the animals showed a preference for the light side of the tank. Furthermore, the observed effect of EO, E1, CIT and GER was reversed by pre-treatment with FMZ, suggesting that GABAergic receptors were involved in the anxiolytic effect displayed by these samples. The association between CIT and GER in the lowest studied concentrations showed an interesting synergistic behavior on anxiolytic effect observed in light-dark test. Besides, it was demonstrated that E1 was constituted by phenolic and flavonoid compounds, which could be involved in the observed effect. CONCLUSION: This work has proved that the low-cost zebrafish can be an adequate alternative as an animal model to evaluate the anxiolytic effect of C. citratus and its related compounds. Moreover, the involvement of GABAA receptors could be responsible for the effect showed by the samples. These obtained results can potentially validate the ethnopharmacological use of C. citratus as a medicinal plant for the treatment of anxiety disorders in folk medicine.

Probing the antioxidant activity of Δ9-tetrahydrocannabinol and cannabidiol in Cannabis sativa extracts.

Herein, we report the antioxidant activity of cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC) in pure and mixed solutions at different ratios, as well as of six different Cannabis sativa extracts containing various proportions of CBD and THC by using spectrophotometric (reducing power assay, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), hypochlorous acid (HOCl) scavenging assays) and electrochemical methods (cyclic voltammetry and differential pulse voltammetry). The isolated cannabinoids, the different stoichiometric ratios of CBD and THC, and the natural extracts proved to have remarkable antioxidant properties in all the methods employed in this work. The antioxidant activity of CBD and THC was compared against that of the well-defined antioxidants such as ascorbic acid (AA), resveratrol (Resv) and (-)-epigallocatechin-3-gallate (EGCG). Clear evidence of the synergistic and antagonistic effects between CBD and THC regarding to their antioxidant activities was observed. Moreover, a good correlation was obtained between the optical and electrochemical methods, which proved that the reported experimental procedures can easily be adapted to determine the antioxidant activity of extracts from various Cannabis sativa species and related compounds.