J. Pharm. Pharmacogn. Res., vol. 11, no. 1, pp. 47-54, January-February 2023.

DOI: https://doi.org/10.56499/jppres22.1507_11.1.47

Original Article

Rhamnetin decreases the expression of HMG-CoA reductase gene and increases LDL receptor in HepG2 cells

[Ramnetina disminuye la expresión del gen de la HMG-CoA reductasa y aumenta los receptores de LDL en las células HepG2]

Raghad R. Al-Yousef¹, Manal M. Abbas^1,2, Razan Obeidat², Manal A. Abbas^1,2*

¹Faculty of Allied Medical Sciences, Al-Ahliyya Amman University, Amman 19328, Jordan.

²Pharmacological and Diagnostic Research Center, Al-Ahliyya Amman University, Amman 19328, Jordan.

*E-mail: m.abbas@ammanu.edu.jo

Abstract

Context: Rhamnetin is a naturally occurring methylated derivative of quercetin. This flavonoid is abundant in Syzygium aromaticum, Coriandrum sativum Prunus cerasus, and Rhamnus spp.

Aims: To evaluate the effects of rhamnetin on HMG-CoA reductase and low-density lipoprotein receptor (LDLR) gene and protein expressions in the HepG2 hepatoma cell line.

Methods: The expression of HMG-CoA reductase and LDLR genes and proteins were studied in HepG2 liver cancer cell line by PCR, Western blot, and indirect ELISA, as well as their antioxidant activity.

Results: Rhamnetin was non-toxic up to 200 μM on HepG2 at 24, 48, and 72 h. Rhamnetin (25 µM) upregulated LDLR gene expression by 1.66 folds compared to 3.12 folds exerted by the well-known hypocholesterolemic drug simvastatin. Rhamnetin (100 µM) increased the expression of LDLR protein at the cell membrane, while the other concentrations produced no significant change from the control (vehicle-treated). In HepG2 cell lysate, LDLR was increased by 50 µM of rhamnetin. Also, rhamnetin increased SOD activity significantly by 100.98, 86.28, and 100.98% by the concentrations 25, 50, and 100 µM, respectively. Using the same concentrations, rhamnetin reduced H₂O₂ levels by 50, 67, and 76.34%, respectively.

Conclusions: This study demonstrated for the first time that rhamnetin reduced HMG-CoA reductase gene expression and increased LDLR in HepG2 cells.

Keywords: HepG2; hydroxymethylglutaryl CoA reductase; LDL; rhamnetin; receptors.

Resumen

Contexto: La ramnetina es un derivado metilado natural de la quercetina. Este flavonoide abunda en las especies Syzygium aromaticum, Coriandrum sativum, Prunus cerasus y Rhamnus spp.

Objetivos: Evaluar los efectos de la ramnetina en las expresiones génicas y proteicas de la HMG-CoA reductasa y el receptor de la lipoproteína de baja densidad (LDLR) en la línea celular de hepatoma HepG2.

Métodos: Se estudió la expresión de los genes y proteínas de la HMG-CoA reductasa y del LDLR en la línea celular de hepatoma HepG2 mediante PCR, Western blot y ELISA indirecto, así como su actividad antioxidante.

Resultados: La ramnetina fue no tóxica hasta 200 μM en HepG2 a las 24, 48 y 72 h. La ramnetina (25 µM) aumentó la expresión del gen LDLR en 1,66 veces en comparación con 3,12 veces ejercida por el conocido fármaco hipocolesterolemiante simvastatina. La ramnetina (100 µM) aumentó la expresión de la proteína LDLR en la membrana celular, mientras que las demás concentraciones no produjeron cambios significativos con respecto al control (tratado con vehículo). En el lisado de células HepG2, el LDLR aumentó con 50 µM de ramnetina. Asimismo, la ramnetina aumentó significativamente la actividad de la SOD en 100,98; 86,28 y 100,98% mediante las concentraciones de 25, 50 y 100 µM, respectivamente. Utilizando las mismas concentraciones, la ramnetina redujo los niveles de H₂O₂ en 50, 67 y 76,34%, respectivamente.

Conclusiones: Este estudio demostró por primera vez que la ramnetina redujo la expresión del gen de la HMG-CoA reductasa y aumentó el LDLR en células HepG2.

Palabras Clave: HepG2; hidroximetilglutaril CoA reductasa; LDL; ramnetina; receptores.

Citation Format: Al-Yousef RR, Abbas MM, Obeidat R, Abbas MA (2023) Rhamnetin decreases the expression of HMG-CoA reductase gene and increases LDL receptors in HepG2 cells. J Pharm Pharmacogn Res 11(1): 47–54. https://doi.org/10.56499/jppres22.1507_11.1.47

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