Journal
AMERICAN JOURNAL OF TRANSPLANTATION
Volume 4, Issue 3, Pages 352-362Publisher
WILEY
DOI: 10.1111/j.1600-6143.2004.00357.x
Keywords
apoptosis; Bax; cytochrome C; ischemia-reperfusion; liver transplantation; mitochondrial calcium
Categories
Funding
- NCI NIH HHS [5P30 CA 68485-07, T32 CA009592] Funding Source: Medline
- NIDDK NIH HHS [1K08 DK 59390-01, T32 DK 07673] Funding Source: Medline
Ask authors/readers for more resources
Mitochondrial calcium (mCa + 2) overload occurs during cold preservation and is an integral part of mitochondrial-dependent apoptotic pathways. We investigated the role of mCa + 2 overload in cell death following hypothermic storage using HepG2 cells stored in normoxic-hypothermic (4 degreesC) or hypoxic (<0.1% O2)- hypothermic Belzer storage solution. Cells were stored for 6 h, with or without 10 l M ruthenium red (mCa + 2 uniporter inhibitor) followed by rewarming in oxygenated media at 37 degrees C. Cytoplasmic cytochrome c levels were studied by Western analysis and by fluorescent microscopy after transfection of cytochrome c-GFP expression plasmid. Immunofluorescence determined the intracellular, spatio-temporal distribution of Bax, and TUNEL staining was used to evaluate cell death after 180 min of rewarming. Caspase activation was evaluated using Western analysis and a caspase 3 activity assay. Bax translocation, cytochrome c release, and early rewarming cell death occurred following hypothermic storage and were exacerbated by hypoxia. Caspase 3 activation did not occur following hypothermic storage. Blockade of mCa + 2 uptake prevented Bax translocation, cytochrome c release, and early rewarming cell death. These studies demonstrate that mCa + 2 uptake during hypothermic storage, both hypoxic and normoxic, contributes to early rewarming apoptosis by triggering Bax translocation to mitochondria and cytochrome c release.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available