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ORIGINAL ARTICLE
Year : 2015  |  Volume : 5  |  Issue : 1  |  Page : 9-16

The assessment of an in-vitro model for evaluating the role of PARP in ethanol-mediated hepatotoxicity


1 Department of Environmental and Occupational Health, College of Public Health, Tampa, Florida, USA
2 Department of Pathology and Cell Biology, Morsani College of Medicine, Tampa, Florida, USA
3 College of Nursing, University of South Florida, Tampa, Florida, USA

Correspondence Address:
R D Harbison
Department of Environmental and Occupational Health, College of Public Health, University of South Florida, Tampa, Florida - 33612
USA
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2229-5151.152300

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This investigation aims to assess whether the hepatocellular carcinoma cell line, HepG2, is an appropriate model to assess the role of poly (ADP-ribose) polymerase (PARP) during acute ethanol toxicosis. HepG2 cells were dosed with graded concentrations of ethanol, ranging from 100 mM to 800 mM, for 6 hours to assess PARP activity induction, while another parallel experiment examined cellular damage via medium aspartate aminotransferase activity and cellular viability via MTT reduction. Aspartate aminotransferase activity was significantly elevated at 600 mM ethanol (FOLD; P < 0.01), with further increases at the 800 mM dose (1.43 fold; P < 0.001), compared to controls. Cellular viability was not significantly decreased compared to controls among all dose groups. PARP activity measured in total cell lysates showed a significant decreasing trend with respect to ethanol dose, reaching statistical significance at the 100 mM dose group (P < 0.05). Paradoxically, exposure to 50 μM etoposide (Positive apoptosis-inducing control) did not demonstrate significant PARP activity ablation. When analyzing PARP activity observation temporally, a significant correlation (R΂ =0.5314) was observed between activity and assay sequence. Overall, a clear HepG2 insensitivity to ethanol was observed.


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