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Molecular Mechanisms of Apoptosis Induced by Ajoene in 3T3-L1 Adipocytes
Article first published online: 6 SEP 2012
2006 North American Association for the Study of Obesity (NAASO)
Volume 14, Issue 3, pages 388–397, March 2006
How to Cite
Yang, J.-Y., Della-Fera, M. A., Nelson-Dooley, C. and Baile, C. A. (2006), Molecular Mechanisms of Apoptosis Induced by Ajoene in 3T3-L1 Adipocytes. Obesity, 14: 388–397. doi: 10.1038/oby.2006.52
- Issue published online: 6 SEP 2012
- Article first published online: 6 SEP 2012
- Received for review September 01, 2005; Accepted in final form December 20, 2005
- reactive oxygen species;
- apoptosis-inducing factor;
- 3T3-L1 adipocytes
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 μM. 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.