Oxalate-induced activation of PKC-alpha and -delta regulates NADPH oxidase-mediated oxidative injury in renal tubular epithelial cells.
Oxalate-induced oxidative stress contributes to cell injury and promotes renal deposition of calcium oxalate crystals. However, we do not know how oxalate stimulates reactive oxygen species (ROS) in renal tubular epithelial cells. We investigated the signaling mechanism of oxalate-induced ROS formation in these cells and found that oxalate significantly increased membrane-associated protein kinase C (PKC) activity while at the same time lowering cytosolic PKC activity. Oxalate markedly translocated PKC-alpha and -delta from the cytosol to the cell membrane. Pretreatment of LLC-PK1 cells with specific inhibitors of PKC-alpha or -delta significantly blocked oxalate-induced generation of superoxide and hydrogen peroxide along with NADPH oxidase activity, LDH release, lipid hydroperoxide formation, and apoptosis. The PKC activator PMA mimicked oxalate's effect on oxidative stress in LLC-PK1 cells as well as cytosol-to-membrane translocation of PKC-alpha and -delta. Silencing of PKC-alpha expression by PKC-alpha-specific small interfering RNA significantly attenuated oxalate-induced cell injury by decreasing hydrogen peroxide generation and LDH release. We believe this is the first demonstration that PKC-alpha- and -delta-dependent activation of NADPH oxidase is one of the mechanisms responsible for oxalate-induced oxidative injury in renal tubular epithelial cells. The study suggests that the therapeutic approach might be considered toward attenuating oxalate-induced PKC signaling-mediated oxidative injury in recurrent stone formers.
Thamilselvan
Vijayalakshmi
V
Vattikuti Urology Institute, Henry Ford Health System, Detroit, Michigan 48202, USA.
Menon
Mani
M
Thamilselvan
Sivagnanam
S
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004847:Epithelial Cells / Q000473:pathology*; D006861:Hydrogen Peroxide / Q000378:metabolism; D007684:Kidney Tubules / Q000473:pathology*; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D018374:LLC-PK1 Cells; D015227:Lipid Peroxidation / Q000187:drug effects; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000502:physiology*; D010070:Oxalates / Q000494:pharmacology*; D018384:Oxidative Stress / Q000502:physiology*; D051571:Protein Kinase C-alpha / Q000037:antagonists & inhibitors*; D051745:Protein Kinase C-delta / Q000037:antagonists & inhibitors*; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors; D015398:Signal Transduction / Q000502:physiology; D013347:Subcellular Fractions / Q000378:metabolism; D013552:Swine; D014162:Transfection
D004791:Enzyme Inhibitors; D010070:Oxalates; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D019255:NADPH Oxidases; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta
Protoplasma. 2001;217(1-3):117-24
11732328
Inflamm Res. 1999 Jul;48(7):412-6
10450792
Urol Res. 2003 Mar;31(1):3-9
12624656
J Biol Chem. 1997 Jun 6;272(23):14950-3
9169467
J Exp Med. 1996 Dec 1;184(6):2399-404
8976194
Am J Physiol Heart Circ Physiol. 2007 Apr;292(4):H1675-84
17209006
Contrib Nephrol. 1993;101:190-3
8467674
Free Radic Biol Med. 2003 Sep 1;35(5):504-16
12927600
Biochem J. 1998 Jun 1;332 ( Pt 2):281-92
9601053
Am J Physiol Renal Physiol. 2005 Jun;288(6):F1227-35
15657300
Mol Pharmacol. 1995 Dec;48(6):995-1003
8848016
Malar J. 2007 Apr 16;6:45
17437631
Cancer Metastasis Rev. 1989 Dec;8(3):199-214
2697470
J Urol. 2002 Jul;168(1):253-9
12050552
J Gen Virol. 2001 Nov;82(Pt 11):2597-2605
11602770
Free Radic Biol Med. 2007 Aug 1;43(3):332-47
17602948
J Urol. 2005 Feb;173(2):640-6
15643280
Am J Physiol Cell Physiol. 2000 Dec;279(6):C1812-8
11078696
BJU Int. 2005 Jul;96(1):117-26
15963133
Int J Biochem Cell Biol. 2007;39(1):44-84
16978905
Physiol Rev. 1979 Jul;59(3):527-605
37532
J Urol. 1996 Mar;155(3):1112-6
8583575
Urol Res. 2005 Feb;33(1):65-9
15565439
J Urol. 1990 Dec;144(6):1535-8
2231959
J Invest Dermatol. 2006 Feb;126(2):460-7
16374459
Scanning Microsc. 1995 Jun;9(2):597-614; discussion 614-6
8714752
J Biol Chem. 2001 Mar 23;276(12):8910-7
11120743
J Am Soc Nephrol. 1999 Nov;10 Suppl 14:S441-5
10541280
Mol Cell. 2003 Jan;11(1):35-47
12535519
Biochemistry. 2002 Jun 18;41(24):7743-50
12056906
Mol Endocrinol. 1999 Dec;13(12):2002-12
10598577
Am J Physiol. 1988 Nov;255(5 Pt 1):C699-704
3189535
J Biol Chem. 1993 May 5;268(13):9194-7
8486620
Am J Nephrol. 2003 Nov-Dec;23(6):380-9
14551462
Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9220-5
9689061
Brain Pathol. 2002 Jan;12(1):21-35
11770899
J Urol. 2000 Jul;164(1):224-9
10840464
J Bone Miner Res. 1995 Dec;10(12):1875-84
8619367
Am J Physiol Regul Integr Comp Physiol. 2003 Aug;285(2):R277-97
12855411
FASEB J. 2003 Nov;17(14):1986-95
14597668
Circ Res. 1999 Aug 6;85(3):264-71
10436169
Immunology. 1993 Nov;80(3):360-6
8288312
Curr Drug Metab. 2008 Oct;9(8):686-96
18855607
Kidney Int. 2005 Dec;68(6):2526-41
16316329
Arterioscler Thromb Vasc Biol. 2002 Dec 1;22(12):2037-43
12482831
J Am Soc Nephrol. 1999 Nov;10 Suppl 14:S457-63
10541283
J Biol Chem. 2002 Dec 13;277(50):48152-7
12368297
Antioxid Redox Signal. 2006 Sep-Oct;8(9-10):1597-607
16987014
Circ Res. 2002 May 17;90(9):1012-9
12016268
Chem Biol Interact. 2000 Dec 1;129(1-2):195-208
11154741
Kidney Int. 1996 Feb;49(2):413-9
8821825
J Urol. 1997 Mar;157(3):1059-63
9072543
Arterioscler Thromb Vasc Biol. 2001 May;21(5):739-45
11348868
Proc Natl Acad Sci U S A. 2006 Sep 5;103(36):13391-6
16938849
Antioxid Redox Signal. 2007 Jan;9(1):49-89
17115887
J Biol Chem. 1999 Feb 5;274(6):3764-71
9920929
J Neurosci Res. 1999 Jun 15;56(6):668-78
10374822
J Urol. 2005 Aug;174(2):757-60
16006971
BJU Int. 2009 Jul;104(1):115-20
19220254
Urol Res. 2005 Nov;33(5):340-8
16284879
J Biol Chem. 2005 Jun 24;280(25):23709-17
15831499
Circ Res. 2005 Sep 2;97(5):482-7
16081872
Biochem Pharmacol. 2006 Jul 14;72(2):176-83
16730666
Biochem Biophys Res Commun. 2004 Jul 30;320(3):789-94
15240117
J Biol Chem. 1951 Nov;193(1):265-75
14907713
J Immunol. 2005 Sep 15;175(6):4009-16
16148149
Am J Nephrol. 2004 Sep-Oct;24(5):557-68
15539792
Circ Res. 1994 Jun;74(6):1141-8
8187280
Biochem Biophys Res Commun. 2002 Nov 15;298(5):738-43
12419315