J Korean Med Sci.  2010 Nov;25(11):1652-1656. 10.3346/jkms.2010.25.11.1652.

Time Dependent Bladder Apoptosis Induced by Acute Bladder Outlet Obstruction and Subsequent Emptying is Associated with Decreased MnSOD Expression and Bcl-2/Bax Ratio

• Li WJ
• Shin MK
• Oh SJ

• KMID: 1779237
• DOI: http://doi.org/10.3346/jkms.2010.25.11.1652

Abstract

Ischemia/reperfusion (I/R) injury-induced oxidative stress plays an important role in the functional impairment of the bladder following acute bladder outlet obstruction (BOO) via induction of apoptosis. The purpose of this study was to investigate the time course of the bladder apoptosis, and apoptosis related molecular changes in the early stage of acute BOO. Twelve-week-old male Sprague Dawley rats were divided into control, acute BOO only (I), and acute BOO plus subsequent emptying (I/R) for 30, 60, 120 min, 3 days and 2 weeks. We examined the extent of bladder apoptosis, expression of Mn-superoxide dismutase (Mn-SOD), Bcl-2, Bax, caspase 3 and poly (ADP-ribose) (PAR) in the bladder. Bladder apoptosis was significantly increased in the I/R group at 30, 60, and 120 min following bladder emptying. BOO plus subsequent emptying for 30, 60, 120 min showed significant decrease in MnSOD and Bcl-2 expression, and significant increase in caspase 3, Bax expression, and amounts of PAR. These results indicate that bladder apoptosis, induced by acute BOO and subsequent emptying, is associated with decreased MnSOD expression, increased PARP activity and imbalance in apoptosis pathways.

Keyword

Urinary Bladder Neck Obstruction; Apoptosis; Oxidative Stress; Poly (ADP-ribose) Polymerases

MeSH Terms

Animals
*Apoptosis
Caspase 3/metabolism
Male
Oxidative Stress
Poly(ADP-ribose) Polymerases/metabolism
Proto-Oncogene Proteins c-bcl-2/*metabolism
Rats
Rats, Sprague-Dawley
Superoxide Dismutase/*metabolism
Time Factors
Urinary Bladder/enzymology/metabolism
Urinary Bladder Neck Obstruction/enzymology/*metabolism
bcl-2-Associated X Protein/*metabolism

Figure

• 

  Fig. 1 Detection of apoptosis: Representative micrographs show TUNEL positive cells as black-brown (magnification ×400), that are indicated by using arrows. Bar graphs show quantitative image analysis. The apoptotic index represents the percent of apoptotic cells within the total number of cells in a given area. *P<0.01 vs control group; †P<0.01 vs ischemia only group. (A): Control group; (B): Overdistension without reperfusion (ischemia only group, I); (C)-(G): Overdistension with reperfusion for 30 min, 60 min, 120 min, 3 days and 2 weeks, respectively (ischemia/reperfusion group, I/R).

• 

  Fig. 2 Western blot analysis: Representative western blot shows expression of MnSOD, caspase 3, PAR, Bcl-2 and Bax. Bar graphs depict quantitative analysis for each protein expression in the bladder. The expression of each protein was normalized to β-actin. *P<0.01 vs control group; †P<0.01 vs ischemia only group. I, overdistension without reperfusion (ischemia only group); I/R 30 min-2 weeks, overdistension with reperfusion for 30 min, 60 min, 120 min, 3 days and 2 weeks, respectively.

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