Partial attenuation of cytotoxicity and apoptosis by SOD1 in ischemic renal epithelial cells

Huan Ling Liang, Jody Arsenault, Jordan Mortensen, Frank Park, Christopher P. Johnson, Vani Nilakantan

Research output: Contribution to journalArticle

16 Citations (Scopus)

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-PK1) 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 (O2 •-) 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 O2 •- 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-PK1 cells with the mitochondrial antioxidant, MitoTEMPO. Treatment with MitoTEMPO also decreased ATP depletion induced cytotoxicity in LLC-PK1 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.

Original languageEnglish (US)
Pages (from-to)1176-1189
Number of pages14
JournalApoptosis
Volume14
Issue number10
DOIs
StatePublished - Oct 1 2009

Fingerprint

Cytotoxicity
Superoxide Dismutase
Epithelial Cells
Apoptosis
Adenosine Triphosphate
Kidney
LLC-PK1 Cells
Caspase 3
Superoxides
Reactive Oxygen Species
Recovery
Superoxide Dismutase-1
Reperfusion Injury
Catalase
Hydrogen Peroxide
Reperfusion
Ischemia
Antioxidants
Chemical activation
Wounds and Injuries

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Pharmaceutical Science
  • Clinical Biochemistry
  • Cell Biology
  • Biochemistry, medical
  • Cancer Research

Cite this

Liang, H. L., Arsenault, J., Mortensen, J., Park, F., Johnson, C. P., & Nilakantan, V. (2009). Partial attenuation of cytotoxicity and apoptosis by SOD1 in ischemic renal epithelial cells. Apoptosis, 14(10), 1176-1189. https://doi.org/10.1007/s10495-009-0393-z

Partial attenuation of cytotoxicity and apoptosis by SOD1 in ischemic renal epithelial cells. / Liang, Huan Ling; Arsenault, Jody; Mortensen, Jordan; Park, Frank; Johnson, Christopher P.; Nilakantan, Vani.

In: Apoptosis, Vol. 14, No. 10, 01.10.2009, p. 1176-1189.

Research output: Contribution to journalArticle

Liang, HL, Arsenault, J, Mortensen, J, Park, F, Johnson, CP & Nilakantan, V 2009, 'Partial attenuation of cytotoxicity and apoptosis by SOD1 in ischemic renal epithelial cells', Apoptosis, vol. 14, no. 10, pp. 1176-1189. https://doi.org/10.1007/s10495-009-0393-z
Liang, Huan Ling ; Arsenault, Jody ; Mortensen, Jordan ; Park, Frank ; Johnson, Christopher P. ; Nilakantan, Vani. / Partial attenuation of cytotoxicity and apoptosis by SOD1 in ischemic renal epithelial cells. In: Apoptosis. 2009 ; Vol. 14, No. 10. pp. 1176-1189.
@article{d13d26836f5f45c4a10fe90c19af2bc6,
title = "Partial attenuation of cytotoxicity and apoptosis by SOD1 in ischemic renal epithelial cells",
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-PK1) 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 (O2 •-) 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 O2 •- 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-PK1 cells with the mitochondrial antioxidant, MitoTEMPO. Treatment with MitoTEMPO also decreased ATP depletion induced cytotoxicity in LLC-PK1 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 = "Liang, {Huan Ling} and Jody Arsenault and Jordan Mortensen and Frank Park and Johnson, {Christopher P.} and Vani Nilakantan",
year = "2009",
month = "10",
day = "1",
doi = "10.1007/s10495-009-0393-z",
language = "English (US)",
volume = "14",
pages = "1176--1189",
journal = "Apoptosis : an international journal on programmed cell death",
issn = "1360-8185",
publisher = "Springer Netherlands",
number = "10",

}

TY - JOUR

T1 - Partial attenuation of cytotoxicity and apoptosis by SOD1 in ischemic renal epithelial cells

AU - Liang, Huan Ling

AU - Arsenault, Jody

AU - Mortensen, Jordan

AU - Park, Frank

AU - Johnson, Christopher P.

AU - Nilakantan, Vani

PY - 2009/10/1

Y1 - 2009/10/1

N2 - 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-PK1) 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 (O2 •-) 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 O2 •- 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-PK1 cells with the mitochondrial antioxidant, MitoTEMPO. Treatment with MitoTEMPO also decreased ATP depletion induced cytotoxicity in LLC-PK1 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.

AB - 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-PK1) 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 (O2 •-) 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 O2 •- 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-PK1 cells with the mitochondrial antioxidant, MitoTEMPO. Treatment with MitoTEMPO also decreased ATP depletion induced cytotoxicity in LLC-PK1 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.

UR - http://www.scopus.com/inward/record.url?scp=69949147576&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=69949147576&partnerID=8YFLogxK

U2 - 10.1007/s10495-009-0393-z

DO - 10.1007/s10495-009-0393-z

M3 - Article

C2 - 19685188

AN - SCOPUS:69949147576

VL - 14

SP - 1176

EP - 1189

JO - Apoptosis : an international journal on programmed cell death

JF - Apoptosis : an international journal on programmed cell death

SN - 1360-8185

IS - 10

ER -