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  • Title: Molecular mechanisms of apoptosis induced by ajoene in 3T3-L1 adipocytes.
    Author: Yang JY, Della-Fera MA, Nelson-Dooley C, Baile CA.
    Journal: Obesity (Silver Spring); 2006 Mar; 14(3):388-97. PubMed ID: 16648609.
    Abstract:
    OBJECTIVE: Determine the biochemical pathways involved in induction of apoptosis by ajoene, an organosulfur compound from garlic. RESEARCH METHODS AND PROCEDURES: Mature 3T3-L1 adipocytes were incubated with ajoene at concentrations up to 200 microM. Viability and apoptosis were quantified using an MTS-based cell viability assay and an enzyme-linked immunosorbent assay for single-stranded DNA (ssDNA), respectively. Intracellular reactive oxygen species (ROS) production was measured based on production of the fluorescent dye, dichlorofluorescein. Activation of the mitogen-activated protein kinases extracellular signal-regulating kinase 1/2 (ERK) and c-Jun-N-terminal kinase (JNK) was shown by Western blot. Western blot was also used to show activation of caspase-3, translocation of apoptosis-inducing factor (AIF) from mitochondria to nucleus, and cleavage of 116-kDa poly(ADP-ribose) polymerase (PARP)-1. RESULTS: Ajoene induced apoptosis of 3T3-L1 adipocytes in a dose- and time-dependent manner. Ajoene treatment resulted in activation of JNK and ERK, translocation of AIF from mitochondria to nucleus, and cleavage of 116-kDa PARP-1 in a caspase-independent manner. Ajoene treatment also induced an increase in intracellular ROS level. Furthermore, the antioxidant N-acetyl-L-cysteine effectively blocked ajoene-mediated ROS generation, activation of JNK and ERK, translocation of AIF, and degradation of PARP-1. DISCUSSION: These results indicate that ajoene-induced apoptosis in 3T3-L1 adipocytes is initiated by the generation of hydrogen peroxide, which leads to activation of mitogen-activated protein kinases, degradation of PARP-1, translocation of AIF, and fragmentation of DNA. Ajoene can, thus, influence the regulation of fat cell number through the induction of apoptosis and may be a new therapeutic agent for the treatment of obesity.
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