Puromycin aminonucleoside induces oxidant-dependent DNA damage in podocytes in vitro and in vivo

被引:90
作者
Marshall, C. B. [1 ]
Pippin, J. W. [1 ]
Krofft, R. D. [1 ]
Shankland, S. J. [1 ]
机构
[1] Univ Washington, Div Nephrol, Seattle, WA 98195 USA
关键词
podocyte; DNA damage; puromycin aminonucleoside nephropathy; reactive oxygen species; proliferation; glomerulosclerosis; MESANGIAL PROLIFERATIVE GLOMERULONEPHRITIS; EXPERIMENTAL MEMBRANOUS NEPHROPATHY; OXIDATIVE STRESS; GLOMERULAR INJURY; MAMMALIAN-CELLS; OXYGEN RADICALS; NUCLEAR-DNA; NEPHROSIS; RATS; PROTEINURIA;
D O I
10.1038/sj.ki.5001965
中图分类号
R5 [内科学]; R69 [泌尿科学(泌尿生殖系疾病)];
学科分类号
1002 ; 100201 ;
摘要
A decline in podocyte number correlates with progression to glomerulosclerosis. A mechanism underlying reduced podocyte number is the podocyte's relative inability to proliferate in response to injury. Injury by the podocyte toxin puromycin aminonucleoside (PA) is mediated via reactive oxygen species (ROS). The precise role of ROS in the pathogenesis of PA-induced glomerulosclerosis remains to be determined. We sought to examine whether PA-induced ROS caused podocyte DNA damage, possibly accounting for the podocyte's inability to proliferate in response to PA. In vitro, podocytes were exposed to PA, with or without the radical scavenger 1,3-dimethyl-2-thiourea (DMTU). In vivo, male Sprague-Dawley rats were divided into experimental groups (n6/group/time point): PA, PA with DMTU, and control, killed at days 1.5, 3, or 7. DNA damage was measured by DNA precipitation, apurinic/apyrimidinic site, Comet, and 8-hydroxydeoxyguanosine assays. Cell cycle checkpoint protein upregulation (by immunostaining and Western blotting), histopathology, and biochemical parameters were examined. DNA damage was increased in cultured podocytes that received PA, but not PA with DMTU. PA exposure activated specific cell cycle checkpoint proteins, with attenuation by DMTU. DNA repair enzymes were activated, providing evidence for attempted DNA repair. The PA-treated animals developed worse proteinuria and histopathologic disease and exhibited more DNA damage than the DMTU pretreated group. No significant apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling staining. A mechanism underlying the lack of podocyte proliferation following PA-induced injury in vitro and in vivo may be ROS-mediated DNA damage, with upregulation of specific cell cycle checkpoints leading to cell cycle arrest.
引用
收藏
页码:1962 / 1973
页数:12
相关论文
共 60 条
  • [31] OLSON JL, 1981, LAB INVEST, V44, P271
  • [32] MICROELECTROPHORETIC STUDY OF RADIATION-INDUCED DNA DAMAGES IN INDIVIDUAL MAMMALIAN-CELLS
    OSTLING, O
    JOHANSON, KJ
    [J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1984, 123 (01) : 291 - 298
  • [33] Podocyte loss and progressive glomerular injury in type II diabetes
    Pagtalunan, ME
    Miller, PL
    JumpingEagle, S
    Nelson, RG
    Myers, BD
    Rennke, HG
    Coplon, NS
    Sun, LM
    Meyer, TW
    [J]. JOURNAL OF CLINICAL INVESTIGATION, 1997, 99 (02) : 342 - 348
  • [34] Cell biology of the glomerular podocyte
    Pavenstädt, H
    Kriz, W
    Kretzler, M
    [J]. PHYSIOLOGICAL REVIEWS, 2003, 83 (01) : 253 - 307
  • [35] EFFECT OF DIETARY ANTIOXIDANTS ON PUROMYCIN AMINONUCLEOSIDE NEPHROTIC SYNDROME
    PEDRAZACHAVERRI, J
    AREVALO, AE
    HERNANDEZPANDO, R
    LARRIVASAHD, J
    [J]. INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY, 1995, 27 (07) : 683 - 691
  • [36] DNA damage is a novel response to sublytic complement CSb-9-induced injury in podocytes
    Pippin, JW
    Durvasula, R
    Petermann, A
    Hiromura, K
    Couser, WG
    Shankland, SJ
    [J]. JOURNAL OF CLINICAL INVESTIGATION, 2003, 111 (06) : 877 - 885
  • [37] MESANGIAL IMMUNE INJURY, HYPERTENSION, AND PROGRESSIVE GLOMERULAR DAMAGE IN DAHL RATS
    RAIJ, L
    AZAR, S
    KEANE, W
    [J]. KIDNEY INTERNATIONAL, 1984, 26 (02) : 137 - 143
  • [38] RICARDO SD, 1994, J AM SOC NEPHROL, V4, P1974
  • [39] REACTIVE OXYGEN SPECIES IN PUROMYCIN AMINONUCLEOSIDE NEPHROSIS - IN-VITRO STUDIES
    RICARDO, SD
    BERTRAM, JF
    RYAN, GB
    [J]. KIDNEY INTERNATIONAL, 1994, 45 (04) : 1057 - 1069
  • [40] Increased oxidative stress and apoptosis in acute puromycin aminonucleoside nephrosis
    Rincon, J
    Romero, M
    Viera, N
    Pedreañez, A
    Mosquera, J
    [J]. INTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY, 2004, 85 (01) : 25 - 33