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  • Title: Oxidative stress-induced apoptosis prevented by Trolox.
    Author: Forrest VJ, Kang YH, McClain DE, Robinson DH, Ramakrishnan N.
    Journal: Free Radic Biol Med; 1994 Jun; 16(6):675-84. PubMed ID: 8070670.
    Abstract:
    The ability of oxidative stress to induce apoptosis (programmed cell death), and the effect of Trolox, a water soluble vitamin E analog, on this induction were studied in vitro in mouse thymocytes. Cells were exposed to oxidative stress by treating them with 0.5-10 microM hydrogen peroxide (H2O2) for 10 min, in phosphate-buffered saline supplemented with 0.1 mM ferrous sulfate. Cells were resuspended in RPMI 1640 medium with 10% serum and incubated at 37 degrees C under 5% CO2 in air. Electron microscopic studies revealed morphological changes characteristic of apoptosis in H2O2-treated cells. H2O2 treatment fragmented the DNA in a manner typical of apoptotic cells, producing a ladder pattern of 200 base pair increments upon agarose gel electrophoresis. The percentage of DNA fragmentation (determined fluorometrically) increased with increasing doses of H2O2 and postexposure incubation times. Pre- or posttreatment of cells with Trolox reduced H2O2-induced DNA fragmentation to control levels and below. The results indicate that oxidative stress induces apoptosis in thymocytes, and this induction can be prevented by Trolox, a powerful inhibitor of membrane damage.
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