Involvement of hypoxia-triggered endoplasmic reticulum stress in outlet obstruction-induced apoptosis in the urinary bladder.
In bladder outlet obstruction (BOO), mechanical stress and ischemia/hypoxia are implicated in structural and functional alterations of the urinary bladder. Because mechanical stress and hypoxia may trigger endoplasmic reticulum (ER) stress, we examined involvement of ER stress in the damage of the bladder caused by BOO. An experimental model of BOO was established in rats by complete ligature of the urethra for 24 h, and bladders were subjected to northern blot analysis and assessment of apoptosis. Isolated urinary bladders and bladder-derived smooth muscle cells (BSMCs) were also exposed to mechanical strain and hypoxia and used for analyses. To examine involvement of ER stress in the damage of the bladder, the effects of a chemical chaperone 4-phenylbutyrate (4-PBA) were evaluated in vitro and in vivo. Outlet obstruction for 24 h induced expression of ER stress markers, GRP78 and CCAAT/enhancer-binding protein-homologous protein (CHOP), in the bladder. It was associated with induction of markers for mechanical stress (cyclooxygenases 2) and hypoxia (vascular endothelial growth factor and glyceraldehyde-3-phosphate dehydrogenase). When isolated bladders and BSMCs were subjected to mechanical strain, induction of GRP78 and CHOP was not observed. In contrast, when BSMCs were exposed to hypoxic stress caused by CoCl2 or thenoyltrifluoroacetone (TTFA), substantial upregulation of GRP78 and CHOP was observed, suggesting involvement of hypoxia in the induction of ER stress. In the bladder subjected to BOO, the number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cells increased in the epithelial cells and BSMCs. Similarly, treatment with TTFA or CoCl2 induced apoptosis of BSMCs, and 4-PBA significantly attenuated ER stress and apoptosis triggered by these agents. Furthermore, in vivo administration with 4-PBA significantly reduced apoptosis in the bladder subjected to BOO. These results suggested that outlet obstruction caused ER stress via hypoxic stress in the bladder and that hypoxia-triggered ER stress may be involved in the induction of apoptosis in BOO.
Sawada
Norifumi
N
Department of Molecular Signaling, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Chuo, Yamanashi, Japan.
Yao
Jian
J
Hiramatsu
Nobuhiko
N
Hayakawa
Kunihiro
K
Araki
Isao
I
Takeda
Masayuki
M
Kitamura
Masanori
M
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015415:Biomarkers / Q000378:metabolism; D002082:Butylamines / Q000494:pharmacology; D004721:Endoplasmic Reticulum; D005260:Female; D006360:Heat-Shock Proteins / Q000378:metabolism; D000860:Hypoxia / Q000150:complications*; D066298:In Vitro Techniques; D018832:Molecular Chaperones / Q000378:metabolism / Q000494:pharmacology; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013314:Stress, Mechanical; D013312:Stress, Physiological / Q000209:etiology* / Q000378:metabolism; D013997:Time Factors; D051742:Transcription Factor CHOP / Q000378:metabolism; D001743:Urinary Bladder / Q000503:physiopathology*; D001748:Urinary Bladder Neck Obstruction / Q000150:complications / Q000378:metabolism / Q000503:physiopathology*; D000091342:Endoplasmic Reticulum Chaperone BiP
D015415:Biomarkers; D002082:Butylamines; D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D051742:Transcription Factor CHOP; C121358:4-phenylbutylamine; C051684:GRP78 protein, rat; D000091342:Endoplasmic Reticulum Chaperone BiP