Original Articles
Regulation of renal tubular cell apoptosis and proliferation after ischemic injury to a solitary kidney,☆☆

https://doi.org/10.1067/mlc.2001.118926Get rights and content

Abstract

The time course and regulation of apoptosis and cellular regeneration after 30 minutes of acute ischemic injury to a single kidney was elucidated in rats at five time points over 20 weeks. The fraction of apoptotic cells was most prominent at 1 day after the insult in the distal tubule (8% ± 4% vs 0% ± 0%, acute renal failure [ARF] vs sham, respectively) and was still elevated at 7 days (2% ± 2% vs 0% ± 0%). At that time, the whole kidney mRNA expression of the apoptosis inhibitory genes bcl-xL and bcl-2, as well as that of the apoptosis promotor bax, was significantly reduced. Immunohistochemistry of kidney specimen showed suppression of bcl-2 in the distal tubule but up-regulation in the proximal tubule, whereas bax protein was more strongly expressed in the distal tubule. Cellular proliferation started at day 1 and continued over the following 20 weeks, leading to severe tubular dilation and kidney failure. These data indicate that differential regulation of bcl-2 family members contributes to the early apoptotic clearance of lethally injured tubular epithelial cells after ischemic injury to a solitary kidney. (J Lab Clin Med 2001;138:343-51)

Section snippets

Rats

Male Sprague-Dawley rats weighing 263 to 328 g were anesthetized with Inactin, 100 mg/g body wt intraperitoneally. The rats were placed on a temperature-regulated table, received a rectal thermo probe, and were kept at 36.5°C during right nephrectomy and 30 minutes of cross-clamping of the left renal artery. A 200-μL sample of tail vein blood was drawn in all animals, at all time points indicated until the animals were killed, for the measurement of serum creatinine with the standardized

Histomorphology and kidney function

Kidney function data and body weight over time are shown in Table I.

. Time course of serum creatinine and body weight of rats subjected to ARF and that of sham-operated rats

Empty CellTime01h1d3d7d5w20w
BodyARF298 ± 17301 ± 11279 ± 15264 ± 13*268 ± 22*389 ± 13*461 ± 15*
 weight (g)Control286 ± 14285 ± 13277 ± 2286 ± 8305 ± 10432 ± 5501 ± 24
SerumARF0.5 ± 0.10.8 ± 04.0 ± 0.1*3.2 ± 0.8*0.9 ± 0.40.5 ± 0.20.8 ± 0.1*
 creatinine (mg/dL)Control0.4 ± 00.5 ± 0.10.4 ± 00.5 ± 0.10.5 ± 0.10.5 ± 0.10.4 ± 0

h, Hours; d, days; w

Discussion

The present study investigated the time course and molecular regulation of apoptotic cell death and cellular proliferation after ischemic injury to a solitary kidney. This model was chosen because it resembles the clinical setting of kidney transplantation and allows the discrimination between ischemia and alloimmune-mediated injury. Recently three articles were published about apoptosis and apoptosis regulation in rodent kidneys subjected to bilateral clamping of the renal arteries. Basile et

Acknowledgements

We thank Miaofen Chou for technical assistance.

References (31)

  • PL Van-Valenberg et al.

    Mannitol as an indispensible constituent of an intraoperative hydration protocol for the prevention of acute renal failure after renal cadaveric transplantation

    Transplantation

    (1987)
  • BM Hall et al.

    Post-transplant acute renal failure in cadaveric renal recipients treated with cyclosporine

    Kidney Int

    (1985)
  • R Oberbauer et al.

    Apoptosis of tubular epithelial cells predicts early allograft dysfunction in renal transplant recipients

    J Am Soc Nephrol

    (1999)
  • M Perez-Fontan et al.

    Outcome of grafts with long-lasting delayed graft function after renal transplantation

    Transplantation

    (1996)
  • DA Shoskes et al.

    The impact of ischemic and immunologic factors on early graft function in pediatric renal transplantation

    Transplantation

    (1990)
  • A Shimizu et al.

    Apoptosis and cell desquamation in repair process of ischemic tubular necrosis

    Virchows Arch B Cell Pathol Incl Mol Pathol

    (1993)
  • RA Zager et al.

    An evaluation of antioxidant effects on recovery from postischemic acute renal failure

    J Am Soc Nephrol

    (1994)
  • S Olsen et al.

    Primary acute renal failure (“acute tubular necrosis”) in the transplanted kidney: morphology and pathogenesis

    Medicine

    (1989)
  • W Lieberthal et al.

    Mechanisms of death induced by cisplatin in proximal tubular epithelial cells: apoptosis vs necrosis

    Am J Physiol

    (1996)
  • D Hockenbery et al.

    Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death

    Nature

    (1990)
  • D Chandler et al.

    Apoptosis and expression of the bcl-2 proto-oncogene in the fetal and adult human kidney: evidence for the contribution of bcl-2 expression to renal carcinogenesis

    Hum Pathol

    (1994)
  • ZN Oltvai et al.

    Checkpoints of dueling dimers foil death wishes

    Cell

    (1994)
  • DH Humes et al.

    Renal tubule cell repair following acute renal injury

    Miner Electrolyte Metab

    (1995)
  • Y Gavrieli et al.

    Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation

    J Cell Biol

    (1992)
  • Z Dong et al.

    Intemucleosomal DNA cleavage triggered by plasma membrane damage during necrotic cell death. Involvement of serine but not cysteinproteases

    Am J Pathol

    (1997)
  • Cited by (35)

    • miR-182-5p Inhibition Ameliorates Ischemic Acute Kidney Injury

      2017, American Journal of Pathology
      Citation Excerpt :

      A dose of 25 mg/kg (of ASO and MM) or 2.5 mg/kg (of ASO) body weight oligonucleotide in a total volume of 500 μL NaCl 0.9% or saline alone was injected via the tail vein into 8- to 12-week-old male C57BL/6 mice (Charles River, Burlington, MA). Power analysis of the rat AKI study was based on the following assumptions: a SD of 0.65 of day 2 creatinine measurements after the ischemic injury was assumed according to the study of Oberbauer et al.11 A sample size of 10 rats is needed to detect a mean difference of 1 mg/dL for the creatinine values between the ASO treatment and the control group with a power of 0.8 and a significance level of 0.05. In case of lost to follow-up (death) all previous observed creatinine values were included in the analysis.

    • Apoptosis and acute kidney injury

      2011, Kidney International
      Citation Excerpt :

      In renal epithelial cells, Bax and Bak are the primary BCL2 members that increase membrane permeability, whereas Bcl-2 and Bcl-XL (B-cell lymphoma-extra large) antagonize ‘membrane attack’ by Bax and Bak. Both metabolic stress in vitro8,48–50 and renal ischemia in vivo1,36,36,51 increase the Bax/Bcl2 ratio, a primary determinant of cell death.52 In renal cells, ischemia activates Bax42,53 and reduces Bcl2,36,54 markedly altering the Bax/Bcl2 ratio in a pro-apoptotic direction.50

    View all citing articles on Scopus

    Supported by the Josef Skoda Funds, the Else Kröner Fresenius Funds, FWF P-12037MED, and NIH-RO1 DK52841.

    ☆☆

    Reprint requests: Rainer Oberbauer, MD, Klinik für Innere Medizin III, Abteilung für Nephrologie und Dialyse, Währinger Gürtel 18-20, A-1090 Vienna, Austria.

    View full text