Effect of age on hydrogen peroxide mediated contraction damage in the male rat bladder.
OBJECTIVE
Recent studies introduced the concept that reactive oxygen species (ROS) may be a major factor in the progressive deterioration of bladder function induced by benign prostatic hyperplasia in men and following partial outlet obstruction in animals. We determined whether bladder contraction sensitivity to ROS changes with aging. Using H2O2 to simulate ROS damage we compared the sensitivity of the contractile responses of bladder smooth muscle isolated from young and elderly rats to H2O2.


METHODS
Male 3-month-old (young) and 12-month-old (elderly) Sprague-Dawley rats were used for this study with 24 per group. Each rat was anesthetized and the bladder was excised. Two longitudinal strips were cut from the bladder body. Each strip was placed in individual 15 ml baths containing oxygenated Tyrode's solution at 37C. Each strip was stimulated at 32 Hz for 20 seconds with pulses 1 millisecond in duration at 80 V. After electrical field stimulation (EFS) the response to 20 microM carbachol and 120 mM KCl was determined. The bath solution was then exchanged for H2O2 solutions of 6 different concentrations including 0%, 0.0625%, 0.125%, 0.25%, 0.5% and 1.0%. After 1 hour of exposure to H2O2 the tissues were washed free of H2O2, and the response to EFS, carbachol and KCl were measured again. At the end of the experiment the strips were weighed and frozen at -70C for malondialdehyde analysis.


RESULTS
The magnitude of the contractile responses of the young and elderly rats to all forms of stimulation were equal. Hydrogen peroxide caused a dose dependent decrease in the contractile responses of bladder strips to all forms of stimulation. Contractile responses to carbachol and KCl were more sensitive to H2O2 than to EFS. Contractile responses of bladder strips isolated from elderly rats were significantly more sensitive to H2O2 damage than strips isolated from young rats. Malondialdehyde generation of bladder strips isolated from elderly rats was significantly greater than those from young rats.


CONCLUSIONS
It is suggested that aging increases the sensitivity of detrusor contraction to oxidative damage.
Aikawa
Ken
K
Division of Basic and Pharmaceutical Sciences, Albany College of Pharmacy, 106 New Scotland Avenue, Albany, NY 12208, USA.
Leggett
Robert  E
R
Levin
Robert M
RM
D000367:Age Factors; D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D016922:Cellular Senescence / Q000187:drug effects* / Q000502:physiology; D046508:Culture Techniques; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation; D006861:Hydrogen Peroxide / Q000633:toxicity*; D007537:Isometric Contraction / Q000187:drug effects* / Q000502:physiology; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D009130:Muscle, Smooth / Q000187:drug effects* / Q000503:physiopathology; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000633:toxicity*; D001743:Urinary Bladder / Q000187:drug effects* / Q000503:physiopathology; D001748:Urinary Bladder Neck Obstruction / Q000503:physiopathology
D017382:Reactive Oxygen Species; D008315:Malondialdehyde; D011189:Potassium Chloride; D002217:Carbachol; D006861:Hydrogen Peroxide