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Glutathione levels determine apoptosis in macrophages
S E Boggs1, T S McCormick, E G Lapetina
1Molecular Cardiovascular Research Center, Case Western Reserve University School of Medicine and University Hospitals of Cleveland, Cleveland, Ohio, 44106-4958, USA.
Abstract:
Spermine NONOate (SpNO, a nitric oxide donor) induced apoptosis and caspase-3 activity in the macrophage cell line RAW 267.4. RES cells that have been derived from RAW 267.4 cells by repeated exposure to lipopolysaccharide and interferon-gamma (LPS/INF-gamma), followed by outgrowth of viable cells, are resistant to apoptosis and caspase-3 activation upon exposure to SpNO. In this study we have determined that RES cells have lower levels of glutathione (GSH) and a higher oxidative state than RAW cells. Subsequently, RAW and RES cells were depleted of GSH by using l-buthionine-[S,R]-sulfoximine (BSO), a specific inhibitor of GSH synthesis. GSH depleted cells did not undergo apoptosis nor demonstrate caspase-3 activity when they were exposed to SpNO. These results suggest that the redox status of the cell is one of the key factors mediating the apoptotic pathway in which glutathione plays a critical role in mediating apoptosis via NO* and reactive oxygen species (ROS).
Insights
Spermine NONOate (SpNO) induces apoptosis in RAW cells but not in resistant RES cells. Glutathione (GSH) depletion prevents SpNO-induced apoptosis, highlighting GSH
Area of Science:
- Cellular and Molecular Biology
- Immunology
- Biochemistry
Background:
- Spermine NONOate (SpNO), a nitric oxide donor, induces apoptosis and caspase-3 activity in RAW 267.4 macrophage cells.
- RES cells, derived from RAW 267.4 cells, exhibit resistance to SpNO-induced apoptosis and caspase-3 activation.
- RES cells possess lower glutathione (GSH) levels and a higher oxidative state compared to RAW cells.
Purpose of the Study:
- To investigate the role of cellular redox status and glutathione in mediating SpNO-induced apoptosis.
- To compare the apoptotic response of RAW and RES cells to SpNO exposure.
- To elucidate the mechanisms underlying SpNO resistance in RES cells.
Main Methods:
- Utilized macrophage cell lines RAW 267.4 and derived RES cells.
- Administered SpNO to induce apoptosis and measured caspase-3 activity.
- Depleted intracellular glutathione (GSH) using l-buthionine-[S,R]-sulfoximine (BSO).
- Assessed cellular oxidative state and GSH levels.
Main Results:
- SpNO treatment induced apoptosis and caspase-3 activity in RAW cells.
- RES cells demonstrated resistance to SpNO-induced apoptosis and caspase-3 activation.
- GSH-depleted RAW and RES cells showed no apoptosis or caspase-3 activity upon SpNO exposure.
- RES cells exhibited lower GSH levels and a higher oxidative state than RAW cells.
Conclusions:
- Cellular redox status is a critical factor in mediating nitric oxide-induced apoptosis.
- Glutathione plays a pivotal role in regulating apoptosis induced by nitric oxide and reactive oxygen species (ROS).
- GSH depletion confers resistance to SpNO-induced apoptosis, suggesting its protective function in the apoptotic pathway.