Genotoxic Effect of Inorganic Arsenic Compounds Using Comet Assay | Chapter 7 | Technological Innovation in Pharmaceutical Research Vol. 2

 Background: Arsenic is a toxicological threat in the environment today since it has been shown to be a human genotoxin and carcinogen.

Aim : The aim of this study was to determine the genotoxic effect of the four arsenic compounds in HepG2 cells by measuring DNA damage, cellular glutathione (GSH) depletion as a measure of oxidative stress and antioxidant agent, as well as apoptotic and necrotic events.

Design of the Study: Arsenic is a toxicological threat in the world today since it has been shown to be a human genotoxin and carcinogen. Arsenate (As+5), sodium arsenite (As+3), arsenic trioxide (As2O3), and dimethyl arsenic acid (DMA+5), a major metabolite of arsenate, were investigated in this project.

Methodology : HepG2 cells (1.5x105/ml) were treated for 24 hours with the four arsenics (10 M). GSH concentration was determined using reverse phase HPLC with fluorescence detection on harvested cells. The Comet assay was used to look for DNA damage in cells. Flow cytometry was used to detect apoptosis and necrosis in cells at the same time.

Results : The DNA damage in cells given DMA or As2O3 was not significantly different from that in control cells. Cells treated with arsenite and arsenate, on the other hand, showed substantial DNA damage (p0.001, p0.05, respectively). When compared to a control group, arsenic compounds increased glutathione levels. However, only arsenite (p0.001) and arsenate (p0.05) were statistically relevant. However, the findings did not show apoptosis or necrosis in viable adherent cells, despite the fact that this was not statistically significant.

Conclusions: When compared to control, all four arsenicals appeared to increase GSH content and DNA damage, with both arsenite and arsenate being significantly different. More research is required to determine the mechanism(s) of arsenical genotoxicity in GSH depletion.

Author (s) Details

Ghazalla Benhusein
Department of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Tripoli University, P.O.Box 13645, Tripoli, Libya.

Elaine Mutch
Toxicology Unit, Department of Environmental Medicine, University of Newcastle upon Tyne, 4th Floor Devonshire Building, NE1 7RU, United Kingdom.

Faith Williams
Toxicology Unit, Department of Environmental Medicine, University of Newcastle upon Tyne, 4th Floor Devonshire Building, NE1 7RU, United Kingdom.

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