J. Pharm. Pharmacogn. Res., vol. 10, no. 3, pp. 486-495, May-June 2022.

DOI: https://doi.org/10.56499/jppres22.1344_10.3.486

Original Article

Chemical composition, in vitro assessment of antioxidant properties and cytotoxicity activity of ethanolic and aqueous extracts of Ajuga orientalis L. (Lamiaceae)

[Composición química, evaluación in vitro de propiedades antioxidantes y actividad citotóxica de extractos etanólicos y acuosos de Ajuga orientalis L. (Lamiaceae)]

Sawsan A. Oran¹, Arwa R. Althaher^2*, Mohammad A. Al Shhab³

¹Department of Biological Sciences, The University of Jordan, Amman, 11942, Jordan.

²Department of Pharmacy, Al-Zaytoonah University of Jordan, Amman, 11733, Jordan.

³Department of Pharmacology, The University of Jordan, Amman, 11942, Jordan.

*E-mail: a.althaher@outlook.com, a.althaher@zuj.edu.jo

Abstract

Context: Ajuga orientalis L. (Lamiaceae) is an aromatic herb used in traditional medicine in Jordan and neighboring countries.

Aims: To evaluate the cytotoxic potential of ethanolic and aqueous extracts from the aerial parts of A. orientalis against three cell lines MCF-7, Caco-2, and HDFa. In addition to assessing the total phenolic and flavonoid contents, antioxidant activity. Further to analyze the phytochemical constituents.

Methods: The phytochemical analysis was performed using gas chromatography/mass spectrometry. The total phenolic (TPC) and flavonoid (TFC) contents were assessed using colorimetric methods. The antioxidant properties of both extracts were assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and reducing power ability. Cytotoxicity was investigated using methyl thiazol tetrazolium (MTT) assay.

Results: The most abundant compounds in the extract were 9-octadecenoic acid, methyl ester, (E)- (27.2%), hexadecanoic acid, methyl ester (12.8%), and methyl stearate (9.6%). The ethanolic extract demonstrated higher TPC and TFC than the aqueous extract. Also, the ethanolic extract showed higher antioxidant activity than aqueous extract using DPPH and the reducing power ability. Furthermore, the aqueous extract of A. orientalis has a potent cytotoxic effect against the Caco-2 cell line (IC₅₀ = 2.059 ± 0.10 μg/mL). In contrast, the ethanolic extract demonstrated a cytotoxic effect against the MCF-7 cell line (IC₅₀ = 59.32 ± 0.04 μg/mL). Both extracts did not exhibit a toxic effect against normal dermal fibroblast cell line (HDFa). Compared to non-cancerous cells, the ethanolic extract of A. orientalis demonstrated high selectivity against MCF-7 cells and limited selectivity against Caco-2 cells. In comparison, the aqueous extract was highly selective against both cancerous cells.

Conclusions: A. orientalis demonstrated antioxidant properties and significant antiproliferative potential against breast and colon cancer. Therefore, additional investigations are needed to study the mechanism of the cytotoxicity for this plant.

Keywords: Ajuga orientalis; antioxidant properties; cytotoxic activity; GC-MS; MTT.

Resumen

Contexto: Ajuga orientalis L. (Lamiaceae) es una hierba aromática utilizada en la medicina tradicional en Jordania y países vecinos.

Objetivos: Evaluar el potencial citotóxico de extractos etanólicos y acuosos de partes aéreas de A. orientalis frente a tres líneas celulares MCF-7, Caco-2 y HDFa. Además de evaluar el contenido total de fenoles y flavonoides, la actividad antioxidante. Además de analizar los componentes fitoquímicos.

Métodos: El análisis fitoquímico se realizó mediante cromatografía de gases/espectrometría de masas. Los contenidos de fenoles y flavonoides totales se evaluaron mediante métodos colorimétricos. Las propiedades antioxidantes de ambos extractos se evaluaron utilizando la actividad de eliminación de radicales 2,2-difenil-1-picrilhidrazilo (DPPH) y la capacidad de poder reductor. La citotoxicidad se investigó mediante el ensayo de metiltiazol tetrazolio (MTT).

Resultados: Los compuestos más abundantes en el extracto fueron ácido 9-octadecenoico, éster metílico, (E)- (27,2%), ácido hexadecanoico, éster metílico (12,8%) y estearato de metilo (9,6%). El extracto etanólico demostró mayor TPC y TFC que el extracto acuoso. Además, el extracto etanólico mostró mayor actividad antioxidante que el extracto acuoso usando DPPH y la capacidad de poder reductor. Además, el extracto acuoso de A. orientalis tiene un potente efecto citotóxico frente a la línea celular Caco-2 (IC₅₀ = 2,059 ± 0,10 μg/mL). En cambio, el extracto etanólico demostró un efecto citotóxico frente a la línea celular MCF-7 (IC₅₀ = 59,32 ± 0,04 μg/mL). Ambos extractos no exhibieron un efecto tóxico contra la línea celular de fibroblastos dérmicos normales (HDFa). En comparación con las células no cancerosas, el extracto etanólico de A. orientalis demostró una alta selectividad frente a las células MCF-7 y una selectividad limitada frente a las células Caco-2. En comparación, el extracto acuoso fue altamente selectivo contra ambas células cancerosas.

Conclusiones: A. orientalis demostró propiedades antioxidantes y un importante potencial antiproliferativo contra el cáncer de mama y colon. Por lo tanto, se necesitan investigaciones adicionales para estudiar el mecanismo de citotoxicidad de esta planta.

Palabras Clave: actividad citotóxica; Ajuga orientalis; GC-MS; MTT; propiedades antioxidantes.

Citation Format: Oran SA, Althaher AR, Al Shhab MA (2022) Chemical composition, in vitro assessment of antioxidant properties and cytotoxicity activity of ethanolic and aqueous extracts of Ajuga orientalis L. (Lamiaceae). J Pharm Pharmacogn Res 10(3): 486–495. https://doi.org/10.56499/jppres22.1344_10.3.486

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