Abstract
INTRODUCTION AND OBJECTIVES: Urocortin (UCN), which is a member of corticotropin-releasing factor (CRF) family peptide, is known to provide cardioprotection by reducing the ischemia-reperfusion (I/R) caused apoptotic cardiomyocyte cells loss. Recently, UCN has been reported to be expressed in male germ cells of human and rat, but only limited data on the role of UCN in germ cells are reported. In this study, we sought to investigate the role of UCN in the apoptosis of germ cells using the experimental testicular I/R model in the rat. METHODS: The left testes of Sprague-Dawley rats were rotated clockwise by 720 for 1 hour, and then the testes were harvested 0, 1, 3, 6 and 24 hours after detorsion (each group; n1⁄45). Realtime quantitative reverse transcription polymerase chain reaction (qRTPCR) and Western Blotting were used to investigate the change of expression levels of UCN. Terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate biotin nick end labeling (TUNEL) staining was performed to evaluate the association between the expression levels of UCN and germ cell apoptosis. In another series, a total of 5 rats underwent intratesticular injection of Astressin, which is a CRF receptor antagonist, 15 minutes before detorsion following the testicular torsion. Then the testes were harvested 3 hours after detorsion, and TUNEL staining was performed to evaluate the direct effect of UCN on the apoptosis of germ cells after I/R injury. RESULTS: The expression levels of UCN increased for 1, 3 and 6 hours after reperfusion, but significantly decreased after 24 hours. In contrast, the number of TUNEL-positive germ cells per seminiferous tubule did not increase after 1, 3 and 6 hours, but significantly increased after 24 hours compared to controls. The injection of Astressin significantly increased the TUNEL-positive germ cells 3 hours after detorsion, which suggests that UCN plays a role in the apoptosis of germ cells. CONCLUSIONS: Our data suggest that UCN is involved in cytoprotection of germ cells after experimental testicular I/R in the rat. Further examination is expected to elucidate the possible mechanisms of UCN-associated cytoprotection in spermatogenesis.
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