Siflinger-Birnboim Alma, Levin Robert M, Hass Martha A
Department of Pharmaceutical Sciences, Albany College of Pharmacy, Albany, New York 12208, USA.
Neurourol Urodyn. 2008;27(6):532-9. doi: 10.1002/nau.20557.
Oxidation of proteins is presumed to contribute to contractile dysfunction associated with partial outlet obstruction (PBOO) of the urinary bladder. The objective of this study was to determine the acute and chronic effects of PBOO on protein oxidation in urinary bladder detrusor smooth muscle (DSM) and mucosa, and to correlate these findings with in vitro contractile function.
Urinary bladders of New Zealand White (NZW) rabbits were obstructed for 1, 3, 7 and 14 days. Proteins were extracted from the bladder tissues and protein carbonyl formation was assessed using immunoblot assays. In vitro contractile response to field stimulation (32 Hz) and carbachol was evaluated in whole bladder strips at the same time points.
Significant elevations in oxidation of DSM proteins were observed in the MW range of 29-65 kDa after 3 days and 14 days of obstruction. No changes in the oxidative status of mucosal proteins were detected as a result of short or long term obstruction. The intermediate filament protein, desmin (53-55 kDa) was detected in obstructed DSM samples in the same MW range as oxidized proteins. A significant decrease in contractile response to field stimulation and carbachol was observed after 1 day and 3-days respectively, and continued to deteriorate through 14 days.
The increase in protein oxidation at 14-days of obstruction correlates with impaired bladder contractility, suggesting that oxidatively modified proteins may contribute to the contractile and biochemical dysfunction associated with PBOO.
蛋白质氧化被认为与膀胱部分出口梗阻(PBOO)相关的收缩功能障碍有关。本研究的目的是确定PBOO对膀胱逼尿肌平滑肌(DSM)和黏膜中蛋白质氧化的急性和慢性影响,并将这些发现与体外收缩功能相关联。
将新西兰白兔(NZW)的膀胱梗阻1、3、7和14天。从膀胱组织中提取蛋白质,并使用免疫印迹分析评估蛋白质羰基的形成。在相同时间点,在整个膀胱条带上评估对场刺激(32Hz)和卡巴胆碱的体外收缩反应。
梗阻3天和14天后,在29-65kDa的分子量范围内观察到DSM蛋白质氧化显著升高。短期或长期梗阻均未检测到黏膜蛋白质氧化状态的变化。在梗阻的DSM样品中检测到中间丝蛋白结蛋白(53-55kDa),其分子量范围与氧化蛋白质相同。分别在1天和3天后观察到对场刺激和卡巴胆碱的收缩反应显著降低,并持续恶化至14天。
梗阻14天时蛋白质氧化的增加与膀胱收缩力受损相关,表明氧化修饰的蛋白质可能导致与PBOO相关的收缩和生化功能障碍。