Faculty Collaboration Database

Partial attenuation of cytotoxicity and apoptosis by SOD1 in ischemic renal epithelial cells. Apoptosis 2009 Oct;14(10):1176-89

Date

08/18/2009

Pubmed ID

19685188

Pubmed Central ID

PMC3146066

DOI

10.1007/s10495-009-0393-z

Scopus ID

2-s2.0-69949147576   17 Citations

Abstract

Reactive oxygen species (ROS) contribute significantly to apoptosis in renal ischemia-reperfusion (IR) injury, however the exact mechanisms are not well understood. We used novel lentiviral vectors to over-express superoxide dismutase 1 (SOD1) in proximal tubular epithelial (LLC-PK(1)) cells and determined effects of SOD1 following ATP depletion-recovery, used as a model to simulate renal IR. SOD1 over-expression partially protected against cytotoxicity (P < 0.001) and decreased superoxide (O(2) (*-)) in ATP depleted cells. The ATP depletion-mediated increase in nuclear fragmentation, an index of apoptosis and activation of caspase-3 was also partially blocked by SOD1 (P < 0.05). However, SOD1 over-expression was insufficient to completely attenuate caspase-3, indicating that ROS other than cytoplasmic O(2) (*-) are involved in ATP depletion mediated injury. To test the contribution of hydrogen peroxide, a subset of enhanced green fluorescent protein (EGFP) and SOD1 (serum free and injured) cells were treated with polyethylene glycol-catalase (PEG-catalase). As expected there was 50% reduction in cytotoxicity and caspase-3 in SOD1 cells compared to EGFP cells; catalase treatment decreased both indices by an additional 28% following ATP depletion. To test the role of mitochondrial derived superoxide, we also treated a subset of LLC-PK(1) cells with the mitochondrial antioxidant, MitoTEMPO. Treatment with MitoTEMPO also decreased ATP depletion induced cytotoxicity in LLC-PK(1) cells in a dose dependant manner. These studies indicate that both SOD1 dependent and independent pathways are integral in protection against ATP depletion-recovery mediated cytotoxicity and apoptosis, however more studies are needed to delineate the signaling mechanisms involved.

Author List

Liang HL, Arsenault J, Mortensen J, Park F, Johnson CP, Nilakantan V

Author

Christopher P. Johnson MD Professor in the Surgery department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Adenosine Triphosphate
Animals
Apoptosis
Caspase 3
Catalase
DNA Fragmentation
Enzyme Activation
Epithelial Cells
Genetic Vectors
Green Fluorescent Proteins
Humans
Hydrogen Peroxide
Ischemia
Kidney Tubules, Proximal
L-Lactate Dehydrogenase
LLC-PK1 Cells
Lentivirus
Piperidines
Reproducibility of Results
Superoxide Dismutase
Superoxide Dismutase-1
Superoxides
Swine
Time Factors