rss
Gut 56:645-654 doi:10.1136/gut.2006.101683
  • Small intestine

p53 independent induction of PUMA mediates intestinal apoptosis in response to ischaemia–reperfusion

  1. Bin Wu1,
  2. Wei Qiu2,
  3. Peng Wang1,
  4. Hui Yu3,
  5. Tao Cheng3,
  6. Gerard P Zambetti4,
  7. Lin Zhang1,
  8. Jian Yu2
  1. 1University of Pittsburgh Cancer Institute, Hillman Cancer Center, Department of Pharmacology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA
  2. 2University of Pittsburgh Cancer Institute, Hillman Cancer Center, Department of Pathology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA
  3. 3University of Pittsburgh Cancer Institute, Hillman Cancer Center, Department of Radiation Oncology, University of Pittsburgh, Pittsburgh, Pennsylvania, USA
  4. 4Department of Biochemistry, St Jude Children’s Research Hospital, Memphis, Tennessee, USA
  1. Correspondence to:
    Dr Jian Yu or Dr Lin Zhang
    Hillman Cancer Center Research Pavilion, 5117 Centre Ave, Pittsburgh, PA 15213, USA; yuj2{at}upmc.edu or zhanglx{at}upmc.edu
  • Accepted 10 October 2006
  • Revised 21 September 2006
  • Published Online First 24 November 2006

Abstract

Background: The small intestine is highly sensitive to ischaemia–reperfusion (I/R) induced injury which is associated with high morbidity and mortality. Apoptosis, or programmed cell death, is a major mode of cell death occurring during I/R induced injury. However, the mechanisms by which I/R cause apoptosis in the small intestine are poorly understood. p53 upregulated modulator of apoptosis (PUMA) is a p53 downstream target and a member of the BH3-only group of Bcl-2 family proteins. It has been shown that PUMA plays an essential role in apoptosis induced by a variety of stimuli in different tissues through a mitochondrial pathway.

Aims: The role of PUMA in I/R induced injury and apoptosis in the small intestine was investigated. The mechanisms by which PUMA is regulated in I/R induced intestinal apoptosis were also studied.

Methods: Ischaemia was induced by superior mesenteric artery occlusion in the mouse small intestine. Induction of PUMA in response to ischaemia alone, or ischaemia followed by reperfusion (I/R), was examined. I/R induced intestinal apoptosis and injury were compared between PUMA knockout and wild-type mice. The mechanisms of I/R induced and PUMA mediated apoptosis were investigated through analysis of caspase activation, cytosolic release of mitochondrial cytochrome c and alterations of the proapoptotic Bcl-2 family proteins Bax and Bak. To determine whether PUMA is induced by reactive oxygen species and/or reactive nitrogen species generated by I/R, superoxide dismutase (SOD) and N-nitro-l-arginine methyl ester (L-NAME) were used to treat animals before I/R. To determine whether p53 is involved in regulating PUMA during I/R induced apoptosis, PUMA induction and apoptosis in response to I/R were examined in p53 knockout mice.

Results:PUMA was markedly induced following I/R in the mucosa of the mouse small intestine. I/R induced intestinal apoptosis was significantly attenuated in PUMA knockout mice compared with that in wild-type mice. I/R induced caspase 3 activation, cytochrome c release, Bax mitochondrial translocation and Bak multimerisation were also inhibited in PUMA knockout mice. SOD or L-NAME significantly blunted I/R induced PUMA expression and apoptosis. Furthermore, I/R induced PUMA expression and apoptosis in the small intestine were not affected in the p53 knockout mice.

Conclusions: Our data demonstrated that PUMA is activated by oxidative stress in response to I/R to promote p53 independent apoptosis in the small intestine through the mitochondrial pathway. Inhibition of PUMA is potentially useful for protecting against I/R induced intestinal injury and apoptosis.

Footnotes

  • Published Online First 23 November 2006

  • Funding: This work was supported in part by the General Motors (GM) Cancer Research Foundation, the V Foundation for Cancer Research (LZ), the Flight Attendant Medical Research Institute (FAMRI) and the Alliance for Cancer Gene Therapy (ACGT) (JY).

  • Competing interests: None.


Free sample
This recent issue is free to all users to allow everyone the opportunity to see the full scope and typical content of Gut.
View free sample issue >>

Don't forget to sign up for content alerts so you keep up to date with all the articles as they are published.

Navigate This Article