Proteinaceous Cytotoxic Component of Allium sativum Induces Apoptosis of INT-407 Intestinal Cells

Abstract

Garlic has long been known for its wide array of therapeutic effects, including hypolipidemic, antihypertensive, antimicrobial, and possibly anticancer effects; conversely, some adverse effects of garlic, such as acute pain and neurogenic inflammation, have also been reported. However, information detailing the toxicological significance of garlic is scarce. In this study, the cytotoxicities of fresh garlic extract (FGE) and boiled garlic extract (BGE) and their underlying toxic mechanisms were investigated using INT-407 intestinal epithelial cells. A brief exposure (20 minutes) to FGE induced a concentration-dependent increase in cell death (37 +/- 2% at 300 mu g/mL), but no cytotoxic effects were induced after exposure to BGE. For FGE, only the high-molecular-mass (> 10-kDa) proteins were associated with cytotoxic effects. FGE-treated cells showed morphological changes such as increased cell rounding and fragmentation, suggesting programmed cell death (apoptosis). Apoptosis of FGE-treated cells was evaluated by observing the fragmented multinuclei stained with Hoechst 33342. From the cell cycle analysis, the increase in hypodiploidic cells and in the G2/M phase cell population suggested not only apoptosis but also cell cycle arrest of FGE-treated cells. Pretreatment with N-acetyl-L-cysteine almost completely prevented FGE-induced cell death, suggesting that reactive oxygen species (ROS) may play a key role in FGE-associated cytotoxicity. Consumption of fresh garlic may be linked to potential cytotoxicity of intestinal cells when ROS scavengers are not present.