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Mammalian cells are equipped with elaborate systems for protection against the toxicity of reactive oxygen and nitrogen species and electrophiles that are constant dangers to the integrity of their DNA. Phase 2 enzymes (e.g., glutathione transferases, NAD(P)H:quinone reductase) and glutathione synthesis are widely recognized as playing major protective roles against electrophilic carcinogens, but their antioxidant functions have attracted far less attention. The cytotoxicities of four oxidative stressors (menadione, tert-butyl hydroperoxide, 4-hydroxynonenal, and peroxynitrite) for human adult retinal pigment epithelial cells (ARPE-19) were quantified by measuring the concentration dependence of cell death and were expressed as the median effect dose (D(m)) for each oxidant. After treatment of ARPE-19 cells for 24 h with 0-5 microM concentrations of sulforaphane (the powerful Phase 2 enzyme inducer isolated from broccoli), the toxicities of the oxidants were markedly reduced as shown by 1.5- to 3-fold increases in D(m) values. The magnitude of protection was a function of the nature of the oxidants and the concentrations of both the oxidants and sulforaphane. Protection was prolonged and persisted for several days after removal of sulforaphane before returning to control levels. The sulforaphane-dependent increases in specific activities of cytosolic quinone reductase and the glutathione levels were highly significantly correlated with the degree of protection as measured by D(m) values. Antioxidant protection was also demonstrated for human HaCaT keratinocytes and L1210 murine leukemia cells. It is therefore highly likely that the multifaceted and prolonged antioxidant protection provided by sulforaphane is a general phenomenon that is mediated through induction of the Phase 2 enzyme response.

proceedings of the national academy of sciences of the united states of america

Powerful and prolonged protection of human retinal pigment epithelial cells, keratinocytes, and mouse leukemia cells against oxidative damage: The indirect antioxidant effects of sulforaphane