Toxicity evaluation of Ruta angustifolia leaves ethanolic extract

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J. Pharm. Pharmacogn. Res., vol. 11, no. 3, pp. 437-447, May-June 2023. DOI: https://doi.org/10.56499/jppres23.1609_11.3.437 Original Article Acute and repeated dose 28-day oral toxicity of Ruta angustifolia Pers. leaves ethanolic extract in Wistar rats [Toxicidad oral aguda y por dosis repetidas durante 28 días del extracto etanólico de hojas de Ruta angustifolia Pers. en ratas Wistar] Tutik … Continue reading Toxicity evaluation of Ruta angustifolia leaves ethanolic extract

J. Pharm. Pharmacogn. Res., vol. 11, no. 3, pp. 437-447, May-June 2023.

DOI: https://doi.org/10.56499/jppres23.1609_11.3.437

Original Article

Acute and repeated dose 28-day oral toxicity of Ruta angustifolia Pers. leaves ethanolic extract in Wistar rats

[Toxicidad oral aguda y por dosis repetidas durante 28 días del extracto etanólico de hojas de Ruta angustifolia Pers. en ratas Wistar]

Tutik Sri Wahyuni1,2*, Hilkatul Ilmi2, Ade Syamsi Kristiaji3, Martiana Candra Dewi3, Hanifah Khairun Nisa2, Achmad Fuad Hafid1,2, Aty Widyawaruyanti1,2

1Department of Pharmaceutical Sciences, Faculty of Pharmacy, Universitas Airlangga, Surabaya, 60115, East Java, Indonesia.

2Center for Natural Product Medicine Research and Development, Institute of Tropical Diseases, Universitas Airlangga, Surabaya, 60115, East Java, Indonesia.

3Undergraduate Program of Faculty of Pharmacy, Universitas Airlangga, Surabaya, 60115, East Java, Indonesia.

*E-mail: tutik-s-w@ff.unair.ac.id; wahyuni.tutiksri@yahoo.com

Abstract

Context: Ruta angustifolia Pers. has been used as a traditional herb among counties. The ethanol extract of R. angustifolia has proven active as an anti-hepatitis C virus in vitro. Due to the potency of this plant, a safety evaluation is necessary.

Aims: To examine the acute and sub-chronic toxicity of ethanol extract of R. angustifolia leaves in rats.

Methods: An acute toxicity study was done by treating 70% ethanol extract of R. angustifolia (doses of 5, 50, 300, and 2000 mg/kg of BW), while the control group received 1% Tween-80. Toxicity signs and mortality of rats for 14 days were observed. While sub-chronic (28-days repeated dose) toxicity was evaluated with 4 groups, both males and females. The three groups were treated with 70% ethanol extract of R. angustifolia at doses of 50, 500, and 1000 mg/kg BW daily for 28 days, and one group served as a control group. Biochemical, hematological, and histopathological parameters were measured after the period of treatment.

Results: The acute toxicity was no demonstrated toxicity signs or mortality effects up to 14 days of the experiment. Therefore, the LD50 value of the extract was estimated to be higher than 2000 mg/kg BW. The sub-chronic assay revealed no mortality and toxicity effects, including behavior parameters, relative body weight, hematology, and biochemical at 50 and 500 mg/kg BW doses. However, a significant change was seen in the testis of all doses and the liver of the male rats treated at 1000 mg/kg BW. Histopathology assay observed the presence of activated Kupffer cells, sinusoidal dilatation, and infiltration of lymphocytes at the portal vein in the liver with 1000 mg/kg BW. Those may indicate possible toxicity in the liver at the dose of 1000 mg/kg BW. However, no toxic effect at the lower doses tested.

Conclusions: These results suggest that the use of Ruta angustifolia leaves ethanolic extract is relatively safe. The application of high doses needs to be evaluated.

Keywords: health; medicine; medicinal plants; Ruta angustifolia; toxicity assay.

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Resumen

Contexto: Ruta angustifolia Pers. se ha utilizado como una hierba tradicional entre los condados. El extracto etanólico de R. angustifolia demostró ser activo como virus anti-hepatitis C por in vitro. Debido a la potencia de esta planta, es necesaria una evaluación de seguridad.

Objetivos: Examinar la toxicidad aguda y subcrónica del extracto etanólico de hojas de R. angustifolia en ratas.

Métodos: Se realizó un estudio de toxicidad aguda tratando extracto etanólico al 70% de R. angustifolia (dosis de 5, 50, 300 y 2000 mg/kg de PC), mientras que el grupo control recibió Tween-80 al 1%. Se observaron signos de toxicidad y mortalidad de ratas durante 14 días. Mientras que se realizó una evaluación de la toxicidad subcrónica (dosis repetida de 28 días) con 4 grupos, tanto de machos como de hembras. Los tres grupos fueron tratados con extracto etanólico al 70% de R. angustifolia en dosis de 50, 500 y 1000 mg/kg PC diariamente durante 28 días, y un grupo sirvió como grupo de control. Los parámetros bioquímicos, hematológicos e histopatológicos se midieron después del período de tratamiento.

Resultados: La toxicidad aguda no demostró signos de toxicidad ni efectos de mortalidad hasta los 14 días del experimento. Por lo tanto, el valor DL50 del extracto se estimó superior a 2000 mg/kg de peso corporal. El ensayo subcrónico no reveló efectos de mortalidad ni toxicidad, incluidos los parámetros de comportamiento, el peso corporal relativo, la hematología y la bioquímica en dosis de 50 y 500 mg/kg de peso corporal. Sin embargo, se observó un cambio significativo en los testículos de todas las dosis y en el hígado de las ratas macho tratadas con 1000 mg/kg de peso corporal. El ensayo de histopatología observó la presencia de células de Kupffer activadas, dilatación sinusoidal e infiltración de linfocitos en la vena portal hepática con 1000 mg/kg BW. Estos pueden indicar una posible toxicidad en el hígado a la dosis de 1000 mg/kg de peso corporal. Sin embargo, no se observaron efectos tóxicos a las dosis más bajas ensayadas.Conclusiones: Estos resultados sugieren que el uso del extracto etanólico de hojas de Ruta angustifolia es relativamente seguro. La aplicación de dosis altas necesita ser evaluada.

Palabras Clave: ensayo de toxicidad; medicamento; plantas medicinales; Ruta angustifolia; salud.

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Citation Format: Wahyuni TS, Ilmi H, Kristiaji AS, Dewi MC, Nisa HK, Hafid AF, Widyawaruyanti A (2023) Acute and repeated dose 28-day oral toxicity of Ruta angustifolia Pers. leaves ethanolic extract in Wistar rats. J Pharm Pharmacogn Res 11(3): 437–447. https://doi.org/10.56499/jppres23.1609_11.3.437
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