Bcl-X(L) translocation in renal tubular epithelial cells in vitro protects distal cells from oxidative stress

Cuttle, L., Zhang, X. J., Endre, Z. H., Winterford, C., & Gobe, G. C. (2001) Bcl-X(L) translocation in renal tubular epithelial cells in vitro protects distal cells from oxidative stress. Kidney International, 59(5), pp. 1779-88.

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Abstract

BACKGROUND: The molecular pathogenesis of different sensitivities of the renal proximal and distal tubular cell populations to ischemic injury, including ischemia-reperfusion (IR)-induced oxidative stress, is not well-defined. An in vitro model of oxidative stress was used to compare the survival of distal [Madin-Darby canine kidney (MDCK)] and proximal [human kidney-2 (HK-2)] renal tubular epithelial cells, and to analyze for links between induced cell death and expression and localization of selected members of the Bcl-2 gene family (anti-apoptotic Bcl-2 and Bcl-X(L), pro-apoptotic Bax and Bad). METHODS: Cells were treated with 1 mmol/L hydrogen peroxide (H2O2) or were grown in control medium for 24 hours. Cell death (apoptosis) was quantitated using defined morphological criteria. DNA gel electrophoresis was used for biochemical identification. Protein expression levels and cellular localization of the selected Bcl-2 family proteins were analyzed (Western immunoblots, densitometry, immunoelectron microscopy). RESULTS: Apoptosis was minimal in control cultures and was greatest in treated proximal cell cultures (16.93 +/- 4.18% apoptosis) compared with treated distal cell cultures (2.28 +/- 0.85% apoptosis, P < 0.001). Endogenous expression of Bcl-X(L) and Bax, but not Bcl-2 or Bad, was identified in control distal cells. Bcl-X(L) and Bax had nonsignificant increases (P> 0.05) in these cells. Bcl-2, Bax, and Bcl-X(L), but not Bad, were endogenously expressed in control proximal cells. Bcl-X(L) was significantly decreased in treated proximal cultures (P < 0.05), with Bax and Bcl-2 having nonsignificant increases (P> 0.05). Immunoelectron microscopy localization indicated that control and treated but surviving proximal cells had similar cytosolic and membrane localization of the Bcl-2 proteins. In comparison, surviving cells in the treated distal cultures showed translocation of Bcl-X(L) from cytosol to the mitochondria after treatment with H2O2, a result that was confirmed using cell fractionation and analysis of Bcl-X(L) expression levels of the membrane and cytosol proteins. Bax remained distributed evenly throughout the surviving distal cells, without particular attachment to any cellular organelle. CONCLUSION: The results indicate that in this in vitro model, the increased survival of distal compared with proximal tubular cells after oxidative stress is best explained by the decreased expression of anti-apoptotic Bcl-X(L) in proximal cells, as well as translocation of Bcl-X(L) protein to mitochondria within the surviving distal cells.

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ID Code: 67203
Item Type: Journal Article
Refereed: Yes
Additional Information: Cuttle, L
Zhang, X J
Endre, Z H
Winterford, C
Gobe, G C
eng
Research Support, Non-U.S. Gov't
2001/04/25 10:00
Kidney Int. 2001 May;59(5):1779-88.
Keywords: Animals, Apoptosis/drug effects, Blotting, Western, Cell Line, Dogs, Epithelial Cells/cytology/drug effects/metabolism, Gene Expression, Genes, bcl-2, Humans, Hydrogen Peroxide/toxicity, Kidney Tubules, Distal/cytology/drug effects/*metabolism, Kidney Tubules, Proximal/cytology/drug effects/metabolism, Microscopy, Immunoelectron, Oxidative Stress, Proto-Oncogene Proteins c-bcl-2/*genetics/metabolism, *Translocation, Genetic, bcl-X Protein
DOI: 10.1046/j.1523-1755.2001.0590051779.x
ISSN: 0085-2538
Divisions: Current > Schools > School of Biomedical Sciences
Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
Copyright Owner: Copyright 2001 Nature Publishing Group.
Deposited On: 27 Feb 2014 02:30
Last Modified: 27 Feb 2014 02:30

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