Chemical composition, in vitro cytotoxic and antioxidant activities of the essential oil of Peruvian Minthostachys mollis Griseb / Composición química, actividades citotóxicas y antioxidantes in vitro del aceite esencial de Minthostachys mollis Griseb peruano

The composition of the essential oil obtained by hydrodistillation from Minthostachys mollis Griseb (Lamiaceae) aerial parts was determined by GC and GC/MS. Menthone (13.2%), pulegone (12.4%), cis-dihydrocarvone (9.8%) and carvacrol acetate (8.8%) were the main essential oil components. The cytotoxic activity of the essential oil was in vitro measured using the MTT colorimetric assay. IC50 values were calculated on healthy non-tumor cells (HEK-293) and three human cancer cell lines (T24, DU-145 and MCF-7). In such latter cells, the estimated values were around 0.2 mg/mL. In addition, the antioxidant activity was determined by interaction with the stable free radical 2,2"-diphenyl-1-picrylhydrazyl. The essential oil was almost devoid of antioxidant activity indicating that its anti-proliferative action relies on other unknown mechanism.

La composición del aceite esencial obtenido por hidrodestilación a partir de partes aéreas de Minthostachys mollis Griseb (Lamiaceae) se determinó mediante GC y GC/MS. Mentona (13.2%), pulegona (12.4%), junto con cis-dihidrocarvona (9.8%) y acetato de carvacrol (8.8%) fueron los principales componentes del aceite esencial. La actividad citotóxica del aceite esencial se midió in vitro utilizando el ensayo colorimétrico MTT tanto en células sanas no tumorales (HEK-293) como en tres líneas celulares de cáncer humano (T24, DU-145 y MCF-7). Los valores de IC50 calculados fueron de alrededor de 0.2 mg/mL. Además, se determinó la actividad antioxidante por su interacción con el radical libre 2,2"-difenil-1-picrilhidrazilo. El aceite esencial tiene baja actividad antioxidante, lo que indica que su acción antiproliferativa depende de otro mecanismo desconocido.

Direct effect of p, p'-DDT on mice liver

ABSTRACT Contact with the pesticide dichlorodiphenyltrichloroethane (p,p′-DDT) can be the cause of various harmful effects in humans, wildlife, and the environment. This pesticide is known to be persistent, lipophilic, resistant to degradation, and bioaccumulive in the environment and to be slowly released into bloodstream. Growing evidence shows that exposure to DDT is linked to type 2 diabetes mellitus. Individuals exposed to elevated levels of DDT and its metabolite have an increased prevalence of diabetes and insulin resistance. To evaluate these possible relationships, experiments were performed on eight-week-old female mice, divided into three groups (n = 10 per group): Group 1 received a vehicle-control intraperitoneal (i.p.) injection of sesame oil; Groups 2 and 3 received an i.p. dose of 50 and 100 µg/g p,p′-DDT respectively, dissolved in sesame oil. All groups were treated once daily for four days. Real-time PCR analysis of several genes was undertaken. Additionally, biochemical parameters and histopathological changes were measured. NQO1, HMOX1, NR1I3 and NR3C1 were up-regulated in DDT-exposed animals compared to the vehicle control group, while only SREBP1 was down-regulated in the 100 µg/g group. MTTP and FABP5, not previously reported for DDT exposure, but involved in regulation of fatty acid fluxes, could also function as biomarkers cross-talking between these signaling pathways. These results suggest that beyond epidemiological data, there is increasing molecular evidence that DDT may mimic different processes involved in diabetes and insulin resistance pathways.

RESUMO O contato com o praguicida diclorodifeniltricloroetano (p, p'-DDT) pode ser a causa de vários efeitos nocivos sobre os seres humanos, animais silvestres e o meio ambiente. Sabe-se de sua característica de bioacumulação, ser altamente persistente no meio ambiente, lipofílico, resistente à degradação e lentamente liberado na corrente sanguínea. Existe uma evidência crescente de que a exposição ao DDT pode ser ligada a Diabetes mellitus tipo 2. Os indivíduos expostos a níveis elevados de DDT e seu metabólito apresentam maior prevalência de diabetes e resistência à insulina. A fim de obter informações sobre essas possíveis relações, camundongos fêmeas de oito semanas de idade foram divididos em três grupos (n = 10 por grupo): Grupo 1 recebeu um veículo de óleo de gergelim via i.p.; os Grupos 2 e 3 receberam, via i.p., 50 e 100 µg/g de p, p'-DDT, respectivamente, dissolvidos em óleo de gergelim. Todos os grupos foram tratados uma vez ao dia durante quatro dias. Além da análise de PCR em Tempo Real de vários genes, os parâmetros bioquímicos e alterações histopatológicas também foram medidos. A expressão gênica do mRNA dos genes NQO1, HMOX1, NR1I3 e NR3C1 foi maior nos animais expostos ao DDT, em comparação ao grupo controle, enquanto a expressão gênica do SREBP1 diminuiu na concentração de 100 µg/g. Os genes MTTP e FABP5 envolvidos na regulação do fluxo de ácidos graxos, embora não estudados quanto à exposição ao DDT, também podem funcionar como biomarcadores de resposta cruzada entre essas vias de sinalização. Esses resultados sugerem que, além de dados epidemiológicos, há cada vez mais evidências moleculares de que o DDT poderia, de fato, imitar diferentes processos que envolvem as rotas de diabetes e de resistência à insulina.

Perfil de la expresión génica en células sanguíneas aisladas de carpinteros expuestos a solventes orgánicos en Sucre (Colombia) / Gene expression profile in blood cells isolated from carpenters exposed to organic solvents in Sucre (Colombia)

Resumen Objetivo: El objetivo de este estudio fue evaluar la expresión de genes asociados con estrés oxidativo, inflamación y daño al ácido desoxirribonucleico (ADN) en trabajadores de carpinterías en Sucre (Colombia). Materiales y métodos: Aleatoriamente fueron seleccionados 41 individuos de sexo masculino: 28 expuestos y 13 controles, con edades entre 32.3±7.9 y 33.2±8.4 años, respectivamente. Se colectaron muestras de sangre periférica y se realizaron análisis hematológicos y de marcadores de daño hepático. En 24 individuos expuestos y 10 controles se realizó análisis de expresión génica para marcadores de estrés oxidativo, inflamación y daño al ADN usando reacción en cadena de la polimerasa en tiempo real. Resultados: Los parámetros hematológicos y de daño hepático estuvieron dentro de los valores de referencia. La expresión génica de la P53 y BCL-2, genes asociados con el daño al ADN, fue significativamente mayor para el grupo expuesto en comparación con el grupo control. Conclusión: En ausencia de cambios en marcadores hematológicos o de daño hepático, personas expuestas a solventes en Sucre tienen niveles de expresión elevados para los genes P53 y BCL-2. Estos genes podrían ser candidatos útiles como biomarcadores moleculares relacionados con la exposición a solventes.

Abstract Objective: The aim of this study was to evaluate the expression of genes related to oxidative stress, inflammation and deoxyribonucleic acid (DNA) damage in carpentry workers from Sucre, Colombia. Materials and methods: 41 male individuals were randomly selected, 28 exposed and 13 controls, with ages 32.3 ± 7.9 and 33.2 ± 8.4 years old, respectively. Peripheral blood samples were collected and used for hematological and liver damage markers analysis. Gene expression analysis for oxidative stress, inflammation and DNA markers was performed using Real-Time Polymerase Reaction on 24 exposed and 10 controls. Results: Hematological parameters and liver damage markers were found within the reference values. Gene expression of P53 and BCL-2, genes related to DNA damage, was significantly greater for the exposed group when compared with the control group. Conclusion: In the absence of hematological or hepatic damage markers, individuals exposed to solvents in Sucre have increased gene expression for P53 and BCL2. These genes may be useful candidates as molecular biomarkers related to solvent exposure.

Toxicity of the essential oil of the cytral chemotype of Lippia alba (Mill.) N. E. Brown

The essential oil (EO) of Lippiaalba (Mill.) N. E. Brown (Verbenaceae) has been traditionally used to treat several diseases. In this study, the acute toxic effects of the citral chemotype of L. alba EO were evaluated in mice. Animals were treated via intraperitoneal receiving the L. alba essential oil at doses between 50 and 2500 mg/kg, and the control group received sesame oil (vehicle). The EO induced dose-dependent neurotoxic effects at doses greater than 1000 mg/kg, including decreased locomotion, motor skills and muscle strength, hypotonia, dyspnea, kyphosis and convulsions. The EO was lethal at a dose of 2500 mg/kg. Animals receiving 1000 mg/kg were euthanized at the end of the treatment period and their blood and livers were collected for analysis. Mice exposed to L. alba EOpresented significantly greater plasma alanine aminotransferase (ALT) activities than the control group. Liver histological changes included mild inflammation, in particular, an increase in nuclear size. Compared to vehicle control group, changes in expressionfor selected genes were significant for FABP5, a fatty acid transport related gene. In summary, the intraperitoneal administration of L. alba EO (citral chemotype) causes neurological damage in mice at doses equal or greater than 1500 mg/kg, whereas at 1000 mg/kg, it generates mild liver damage. Therefore, the systemic use of this EO raises concerns about its safety.

El aceite esencial (AE) de Lippia alba(Mill.) NE Brown (Verbenaceae) ha sido utilizado tradicionalmente para tratar varias enfermedades. En este estudio, los efectos tóxicos agudos del AE de Lippia alba quimiotipo citral fueron evaluados en ratones. Los animales fueron tratados por vía intraperitonealrecibiendo el AE en dosis entre 50 y 2500 mg/kg de peso, y el grupo control aceite de sésamo (vehículo). Dosis superiores a 1000 mg/kg del AE mostraron efectos neurotóxicos incluyendo disminución de la locomoción e hipotonía, disnea, cifosis y convulsiones. El AE fue letal a la dosis de 2500 mg/kg. Veinticuatro horas después de que los animales fueron tratados con 1000 mg/kg del AE se les realizó eutanasia y su sangre e hígado fueron recolectados para análisis. Los ratones expuestos al AE de L. alba, presentaron actividad alanina aminotransferasa (ALT) en plasma significativamente mayor que el grupo control. Dentro de los cambios histológicos hepáticos se incluyen inflamación leve, en particular, un aumento del tamaño nuclear. En comparación con el grupo control, la expresión de genes seleccionados tuvo diferencias significativas para FABP5, un gen relacionado con el transporte de ácidos grasos. En conclusión, la administración intraperitoneal del AE de L. alba (quimiotipo citral) causa dañosneurológicos en ratones a una dosis igual o superior a 1500 mg/kg, mientras que a 1000 mg/kg, genera daño hepático leve. Por lo tanto, el uso sistémico de este AE plantea preocupaciones en cuanto a su seguridad.