Mechanism of Azaline B-induced Apoptosis in Rat Testis / 대한비뇨기과학회지
Korean Journal of Urology
;
: 750-760, 2005.
Article
in Korean
| WPRIM
| ID: wpr-61279
ABSTRACT
PURPOSE:
To identify the mechanism of azaline B-dependent apoptosis, the regulation of Fas and FasL genes has been investigated. MATERIALS ANDMETHODS:
Azaline B was subcutaneously injected into Sprague-Dawley rats. The levels of Fas receptor (Fas) and Fas ligand (FasL) were detected by reverse transcription-polymerase chain reaction (RT- PCR). Azaline B-dependent apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick and labeling (TUNEL) and DNA fragmentation assay. Transacting factor of FasL promoter was identified by DNase I footprinting and DNA mobility shift assay.RESULTS:
The azaline B-treated testis (250microgram/kg body wt/day) had decreased to 70+/-2.5% and 38+/-1.8% of the normal testis weight at 3 and 5 days after the injection, respectively, but the weights of the testis were not changed after pretreatment of follicle-stimulating hormone (FSH) and testosterone. Apoptosis of the testis was detected by DNA fragmentation assay and TUNEL assay after the azaline B treatment. The levels of Fas and FasL mRNA were increased by the treatment of azaline B in both time- and dose-dependent manners. In DNase I footprinting assay with FasL promoter, the nuclear factor prepared from control was bound with at least four sites SP-1 binding site at 283, NF-kappa B binding site at 219, TATA at 132 and the gamma-interferon response element (gamma-IRE) at 78. gamma-IRE was completely protected by the nuclear extract prepared from azaline B-treated rat testis. In DNA mobility shift assay, the binding activity of gamma-IRE binding protein was increased after azaline B treatment.CONCLUSIONS:
These results suggest that Fas-FasL system may be important to azaline B-dependent apoptosis in rat testis and that gamma-IRE binding protein is related to the azaline B-dependent regulation of FasL gene.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Testis
/
Testosterone
/
Weights and Measures
/
Binding Sites
/
DNA
/
RNA, Messenger
/
Carrier Proteins
/
NF-kappa B
/
Interferon-gamma
/
Rats, Sprague-Dawley
Type of study:
Prognostic study
Limits:
Animals
Language:
Korean
Journal:
Korean Journal of Urology
Year:
2005
Type:
Article
Similar
MEDLINE
...
LILACS
LIS