title: DNA damage response in renal ischemia-reperfusion and ATP-depletion injury of renal tubular cells.
abstract: Renal ischemia-reperfusion leads to acute kidney injury (AKI) that is characterized pathologically by tubular damage and cell death, followed by tubular repair, atrophy and interstitial fibrosis. Recent work suggested the possible presence of DNA damage response (DDR) in AKI. However, the evidence is sketchy and the role and regulation of DDR in ischemic AKI remain elusive. In this study, we demonstrated the induction of phosphorylation of ATM, H2AX, Chk2 and p53 during renal ischemia-reperfusion in mice, suggesting DDR in kidney tissues. DDR was also induced in vitro during the recovery or "reperfusion" of renal proximal tubular cells (RPTCs) after ATP depletion. DDR in RPTCs was abrogated by supplying glucose to maintain ATP via glycolysis, indicating that the DDR depends on ATP depletion. The DDR was also suppressed by the general caspase inhibitor z-VAD and the overexpression of Bcl-2, supporting a role of apoptosis-associated DNA damage in the DDR. N-acetylcysteine (NAC), an antioxidant, suppressed the phosphorylation of ATM and p53 and, to a less extent, Chk2, but NAC increased the phosphorylation and nuclear foci formation of H2AX. Interestingly, NAC increased apoptosis, which may account for the observed H2AX activation. Ku55933, an ATM inhibitor, blocked ATM phosphorylation and ameliorated the phosphorylation of Chk2 and p53, but it increased H2AX phosphorylation and nuclear foci formation. Ku55933 also increased apoptosis in RPTCs following ATP depletion. The results suggest that DDR occurs during renal ischemia-reperfusion in vivo and ATP-depletion injury in vitro. The DDR is partially induced by apoptosis and oxidative stress-related DNA damage. ATM, as a sensor in the DDR, may play a cytoprotective role against tubular cell injury and death.
authors.affiliation: Department of Cellular Biology and Anatomy, Medical College of Georgia, Georgia Reagents University and Charlie Norwood VA Medical Center, Augusta, GA 30912, USA.
authors.forename: Zhengwei
authors.initials: Z
authors.lastname: Ma
authors.affiliation: Department of Cellular Biology and Anatomy, Medical College of Georgia, Georgia Reagents University and Charlie Norwood VA Medical Center, Augusta, GA 30912, USA.
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Wei
authors.affiliation: Department of Cellular Biology and Anatomy, Medical College of Georgia, Georgia Reagents University and Charlie Norwood VA Medical Center, Augusta, GA 30912, USA.
authors.forename: Guie
authors.initials: G
authors.lastname: Dong
authors.affiliation: Vascular Biology Center, Medical College of Georgia, Georgia Reagents University and Charlie Norwood VA Medical Center, Augusta, GA 30912, USA.
authors.forename: Yuqing
authors.initials: Y
authors.lastname: Huo
authors.affiliation: Department of Cellular Biology and Anatomy, Medical College of Georgia, Georgia Reagents University and Charlie Norwood VA Medical Center, Augusta, GA 30912, USA; Department of Nephrology, The Second Xiangya Hospital, Central South University, Changsha, China. Electronic address: zdong@gru.edu.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Dong
mesh_terms: D000111:Acetylcysteine / Q000378:metabolism; D058186:Acute Kidney Injury / Q000235:genetics* / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D064007:Ataxia Telangiectasia Mutated Proteins / Q000235:genetics / Q000378:metabolism; D064447:Checkpoint Kinase 2 / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D005947:Glucose / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000098:blood supply* / Q000378:metabolism; D007684:Kidney Tubules / Q000098:blood supply / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010766:Phosphorylation; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D015424:Reperfusion; D015427:Reperfusion Injury / Q000235:genetics* / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism
chemical_list: D000111:Acetylcysteine; D000255:Adenosine Triphosphate; D064007:Ataxia Telangiectasia Mutated Proteins; D064447:Checkpoint Kinase 2; D005947:Glucose; D006657:Histones; D019253:Proto-Oncogene Proteins c-bcl-2; D016159:Tumor Suppressor Protein p53; C052617:Bcl2 protein, mouse; C576325:Atm protein, mouse; C578633:Chek2 protein, mouse; C475961:gamma-H2AX protein, mouse
keywords: ATP depletion; Acute kidney injury; Apoptosis; DNA damage response; Renal ischemia–reperfusion
references.citation: Methods Enzymol. 1990;186:1-85
references.pmid: 2172697
references.citation: Trends Biochem Sci. 2012 Jan;37(1):15-22
references.pmid: 22079189
references.citation: Biochem Biophys Res Commun. 2012 Feb 10;418(2):267-72
references.pmid: 22260947
references.citation: EMBO J. 2011 Feb 2;30(3):546-55
references.pmid: 21157431
references.citation: Kidney Int. 2012 Dec;82(12):1271-83
references.pmid: 22854643
references.citation: Nat Rev Nephrol. 2011 Apr;7(4):189-200
references.pmid: 21364518
references.citation: Oncogene. 1998 Dec 31;17(26):3401-15
references.pmid: 10030664
references.citation: Toxicol Sci. 2001 Mar;60(1):92-102
references.pmid: 11222876
references.citation: Cell Signal. 2012 May;24(5):981-90
references.pmid: 22286106
references.citation: J Am Soc Nephrol. 2003 Jan;14(1):128-38
references.pmid: 12506145
references.citation: Kidney Int. 2013 Jul;84(1):138-48
references.pmid: 23466994
references.citation: Mol Carcinog. 2010 Jan;49(1):68-74
references.pmid: 19722195
references.citation: Oncogene. 1998 Dec 24;17(25):3341-9
references.pmid: 9916996
references.citation: Am J Pathol. 1992 Apr;140(4):831-8
references.pmid: 1562048
references.citation: Am J Physiol Renal Physiol. 2001 Nov;281(5):F948-57
references.pmid: 11592952
references.citation: Trends Biochem Sci. 2011 Mar;36(3):133-40
references.pmid: 20947357
references.citation: PLoS One. 2012;7(3):e32503
references.pmid: 22442667
references.citation: J Am Soc Nephrol. 2013 Nov;24(11):1720-6
references.pmid: 24136922
references.citation: Asian J Androl. 2013 Mar;15(2):290-6
references.pmid: 23353715
references.citation: Am J Pathol. 2003 Aug;163(2):663-71
references.pmid: 12875985
references.citation: J Biol Chem. 2004 Nov 26;279(48):50455-64
references.pmid: 15375156
references.citation: Gene. 2004 Aug 4;337:1-13
references.pmid: 15276197
references.citation: Oncogene. 2006 Jul 6;25(29):4056-66
references.pmid: 16491117
references.citation: J Biol Chem. 2011 Mar 25;286(12):10411-8
references.pmid: 21285353
references.citation: FASEB J. 2000 May;14(7):955-67
references.pmid: 10783150
references.citation: Nephron Exp Nephrol. 2012;120(3):e103-13
references.pmid: 22678593
references.citation: Biochem Pharmacol. 2003 May 15;65(10):1593-601
references.pmid: 12754095
references.citation: J Clin Invest. 2011 Nov;121(11):4210-21
references.pmid: 22045571
references.citation: Antioxid Redox Signal. 2002 Dec;4(6):915-24
references.pmid: 12573140
references.citation: Endocrinology. 2013 Nov;154(11):4068-77
references.pmid: 24008345
references.citation: Nephrol Dial Transplant. 2010 May;25(5):1435-43
references.pmid: 20037173
references.citation: Nat Rev Mol Cell Biol. 2013 Apr;14(4):197-210
references.pmid: 23847781
references.citation: Methods Mol Biol. 2012;920:613-26
references.pmid: 22941631
references.citation: Methods Mol Biol. 2009;523:161-8
references.pmid: 19381940
references.citation: EMBO J. 2008 Jul 23;27(14):1953-62
references.pmid: 18596698
references.citation: Science. 1998 Dec 4;282(5395):1893-7
references.pmid: 9836640
references.citation: J Am Soc Nephrol. 2014 Oct;25(10):2278-89
references.pmid: 24700871
references.citation: Oncogene. 2001 Apr 5;20(15):1803-15
references.pmid: 11313928
references.citation: J Clin Invest. 2009 May;119(5):1275-85
references.pmid: 19349686
references.citation: EMBO J. 2003 Oct 15;22(20):5612-21
references.pmid: 14532133
references.citation: DNA Repair (Amst). 2004 Aug-Sep;3(8-9):1109-15
references.pmid: 15279799
references.citation: J Neurosci. 1996 Nov 1;16(21):6795-806
references.pmid: 8824320
references.citation: Am J Nephrol. 2010;31(2):178-88
references.pmid: 20016144
references.citation: J Biol Chem. 2008 Mar 7;283(10):6572-83
references.pmid: 18162465
references.citation: Am J Physiol Renal Physiol. 2010 May;298(5):F1078-94
references.pmid: 20200097
references.citation: Ann N Y Acad Sci. 1996 Sep 30;793:427-30
references.pmid: 8906186
references.citation: DNA Repair (Amst). 2003 Feb 3;2(2):211-29
references.pmid: 12531391
references.citation: Kidney Int. 2012 Sep;82(5):516-24
references.pmid: 22673882
references.citation: Acta Otolaryngol. 2008 Apr;128(4):440-5
references.pmid: 18368580
references.citation: Nat Med. 2010 May;16(5):535-43, 1p following 143
references.pmid: 20436483
references.citation: Am J Physiol Renal Physiol. 2013 Dec 1;305(11):F1617-27
references.pmid: 24107420
references.citation: J Am Soc Nephrol. 2009 Aug;20(8):1754-64
references.pmid: 19470675
references.citation: Cancer Res. 2001 Mar 1;61(5):1849-54
references.pmid: 11280737
references.citation: Biochim Biophys Acta. 2013 Dec;1832(12):1998-2008
references.pmid: 23851027
references.citation: Am J Physiol Renal Physiol. 2012 Dec 1;303(11):F1487-94
references.pmid: 22993069
references.citation: Curr Opin Cell Biol. 2004 Dec;16(6):629-33
references.pmid: 15530773
references.citation: Annu Rev Biochem. 2004;73:39-85
references.pmid: 15189136
references.citation: Mol Pharmacol. 2008 Nov;74(5):1292-307
references.pmid: 18703668
references.citation: Nat Commun. 2011 Dec 20;2:593
references.pmid: 22186889