Prooxidant activity and toxicity of nordihydroguaiaretic acid in clone-9 rat hepatocyte cultures

Abstract

Nordihydroguaiaretic acid (NDGA) is a polyphenol. It is present at high concentrations in the leaves of the evergreen desert shrub, Larrea tridentate (Creosote bush), which has a long history of medicinal use traditionally by the native Americans and Mexicans. It is generally believed that the antioxidant properties of NDGA are responsible for the medicinal value of this desert shrub. The clone-9 rat hepatocyte cultures were used as an in vitro model to assess the hepatotoxic potential of NDGA and to determine whether it exhibits any prooxidant activity. The hepatocyte cultures were treated with NDGA for 2 h at 37 degrees C at concentrations of 0-100 microM. After the treatment period the cells, the culture supernatants and cell lysates were assayed for evaluation of prooxidant activity and toxicity of NDGA. Oxidative stress level and oxidative cell injury as measured by the peroxidation of membrane lipids and DNA double-strand breaks were used to index prooxidant activity. Cytotoxicity as measured by the leakage of the liver enzyme lactate dehydrogenase (LDH) into the culture medium, mitochondrial function and extent of cell proliferation were used as the endpoints of toxicity. Significant concentration-dependent differences were observed in these biomarkers over the concentration range examined demonstrating the prooxidant activity and toxicity of NDGA in clone-9 rat hepatocyte cultures.

Research Strategies in the Study of the Pro-Oxidant Nature of Polyphenol Nutraceuticals

“…Using rat hepatocyte cultures, Sahu et al [41] demonstrated concentration-dependent lipid peroxidation upon exposure to nordihydroguaiaretic acid. Similarly, lipid peroxidation of freshly isolated lymphocytes was observed upon treatments with caffeic acid and gallic acid [42] and of erythrocytes by G. biloba extract [43].…”

Rat liver clone‐9 cells in culture as a model for screening hepatotoxic potential of food‐related products: hepatotoxicity of deoxynivalenol

“…In vitro models provide well-defined and reproducible experimental conditions for evaluation of toxicity. They are excellent systems to study mechanisms of action and structure-activity in vitro model for testing the hepatotoxic potential of chemicals and drugs (Barhoumi and Burghardt, 1996;Behrooz and Ismail-Beigi, 1997;Hutchinson et al, 1998;Thompson et al, 1998;Prasad and Ismail-Beigi, 1999;Barhoumi et al, 2000;Liegro et al, 2000;Barros et al, 2001;Garcia-Caballero et al, 2001;Grune et al, 2001Grune et al, , 2002Reeves et al, 2001;Barhoumi et al, 2002;Boess et al, 2003;Neerman et al, 2004;Tsai et al, 2005;Sahu et al, 2006;Vansant et al, 2006). Like normal hepatocytes these cells induce P450 activity (Barhoumi et al, 2000;Reeves et al, 2001).…”

Uso potencial de extractos de Larrea divaricata con bajo contenido de NDGA como antioxidantes en comidas

“…The SOD activity observed at low concentrations of the extract (A), could be related to the presence of other antioxidant compounds. The pro-oxidant activity of NDGA has been proved by other authors, who observed this activity on a clone-9 rat hepatocyte culture, depending on concentrations and the biological environment (Sahu & Ruggles 2006).…”