Toxicogenic effects of Terminalia phanerophlebia on HEK293 cells



J Pharm Pharmacogn Res 9(3): 261-271, 2021.

Original Article

The toxicogenic effect of Terminalia phanerophlebia Engl. & Diels leaf extract on oxidative stress parameters in an in vitro Hek293 model

[Efecto toxicogénico de extracto de hoja de Terminalia phanerophlebia Engl. & Diels sobre parámetros de estrés oxidativo en un modelo in vitro con Hek293]

Marcilyn R. Nyahada1, Daniel G. Amoako1,2*,Anou M. Somboro1,2, Isaiah Arhin1, Hezekiel M. Khumalo1, Rene B. Khan1**

1Drug and Innovation Research Unit, Discipline of Medical Biochemistry, School of Laboratory Medicine and Medical Science, University of KwaZulu-Natal, Durban, South Africa.

2Biomedical Resource Unit, School of Laboratory Medicine and Medical Sciences, College of Health Sciences, University of KwaZulu-Natal; Durban, South Africa.

*E-mail: *amoakodg@gmail.com; **myburgr@ukzn.ac.za

Abstract

Context: Medicinal plants are a highly sought-after alternative to current pharmaceutical drugs because they can be locally cultivated, inexpensive and possess minimal adverse effects. Given that Terminalia phanerophlebia (TP) possesses many useful properties and plays a role in modulating lethal diseases, the cytotoxic effect should be evaluated before its application for therapeutic use.

Aims: To investigate the oxidative effect and molecular mechanisms of TP on human embryonic kidney (HEK293) cells.

Methods: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and adenosine triphosphate (ATP) assays were used to determine the cell viability whilst the thiobarbituric acid reactive species (TBARS) assay was used to detect lipid peroxidation. Endogenous antioxidants, catalase, superoxide dismutase, glutathione peroxidase, heat shock protein 70 and nuclear factor erythroid 2-related factor 2 (Nrf2), were used as oxidative stress markers and were detected via western blotting.

Results: A decrease in cell viability with an IC50 of 1.36 mg/mL and ATP were noted. The concentration of malondialdehyde (MDA) increased significantly (p<0.005). Superoxide dismutase, Nrf2 and heat shock protein concentrations were increased. However, glutathione, glutathione peroxidase and catalase were depleted.

Conclusions: The results obtained suggest that Terminalia phanerophlebia extract is toxicogenic and induced oxidative stress in HEK293 cells.

Keywords: antioxidants; chronic kidney diseases; cytotoxicity; Hek293 cells; lipid peroxidation; oxidative stress; Terminalia phanerophlebia.

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Resumen

Contexto: Las plantas medicinales son una alternativa muy buscada a los medicamentos farmacéuticos actuales porque pueden cultivarse localmente, son económicas y tienen efectos adversos mínimos. Dado que Terminalia phanerophlebia (TP) posee muchas propiedades útiles y juega un papel en la modulación de enfermedades letales, el efecto citotóxico debe evaluarse antes de su aplicación para uso terapéutico.

Objetivos: Investigar el efecto oxidativo y los mecanismos moleculares de TP en células de riñón embrionario humano (HEK293).

Métodos: Se utilizaron ensayos de bromuro de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolio (MTT) y trifosfato de adenosina (ATP) para determinar la viabilidad celular mientras que el ensayo de especies reactivas con ácido tiobarbitúrico (TBARS) utilizado para detectar la peroxidación de lípidos. Se utilizaron antioxidantes endógenos, catalasa, superóxido dismutasa, glutatión peroxidasa, proteína 70 de choque térmico y factor 2 relacionado con el factor nuclear eritroide 2 (Nrf2), como marcadores de estrés oxidativo y se detectaron mediante transferencia Western.

Resultados: Se observó una disminución de la viabilidad celular con una CI50 de 1,36 mg/mL y ATP. La concentración de malondialdehído (MDA) aumentó significativamente (p<0,005). Se incrementaron las concentraciones de superóxido dismutasa, Nrf2 y proteína de choque térmico. Sin embargo, se agotaron glutatión, glutatión peroxidasa y catalasa.

Conclusiones: Los resultados obtenidos sugieren que el extracto de Terminalia phanerophlebia es toxicogénico e induce estrés oxidativo en células HEK293.

Palabras Clave: antioxidantes; células Hek293; citotoxicidad; enfermedades renales crónicas; estrés oxidativo; peroxidación lipídica; Terminalia phanerophlebia.

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Citation Format: Nyahada MR, Amoako DG, Somboro AM, Arhin I, Khumalo HM, Khan RB (2021) The toxicogenic effect of Terminalia phanerophlebia Engl. & Diels leaf extract on oxidative stress parameters in an in vitro Hek293 model. J Pharm Pharmacogn Res 9(3): 261–271.

© 2021 Journal of Pharmacy & Pharmacognosy Research (JPPRes)