Protective effect of osthole on testicular ischemia/reperfusion injury in rats

Testicular torsion is a urological emergency that requires urgent surgical intervention which results in testicular loss if not diagnosed and treated in a timely fashion. Ischemic tissue damage with oxygen deficiency, which starts with the decrease in blood flow to the tissue, continues to increase with the reoxygenation of the damaged tissues as soon as reperfusion is achieved. In various studies, osthole has also been shown to reduce cerebral, spinal cord, intestinal, renal, and myocardial ischemia/perfusion (I/R) damage. The aim of this study is to examine the effects of osthole on testicular I/R injury.

28 Wistar-albino rats were randomly divided into four experimental groups (n=7). Group 1 was the sham operation group. In Group 2 (I/R), 3-h ischemia was created by rotating the testis 720° clockwise, followed by 3 h of reperfusion. In Group 3 (I/R + single dose of Osthole), 20 mg/kg ostol was administered intraperitoneally half an hour before detorsion after 3 h of torsion. The testis was detorsioned. Three h of detorsion was applied. In Group 4 (I/R + twice doses of Osthole), 20 mg/kg ostol was administered intraperitoneally half an hour before detorsion, followed by 3-h torsion. The testis was released and detorsioned. Half an hour after the detorsion, an intraperitoneal dose of 20 mg/kg osthole was administered again. Detorsion was done for 3 h. All rats were sacrificed after 6 h and right orchiectomy was performed for blood for biochemical analysis and histopathological sample.

Glutathion, nuclear respiratory factor 2, Superoxide dismutase, and 8-hydroxydeoxyguanosine levels were decreased in I/R rats, while interleukin-6, malondialdehyde, and myeloperoxidase levels were increased. While caspase 3, caspase 8, caspase 9, and TUNEL showed moderate immunopositive tissues immunohistochemically in rats with I/R damage, mild immunopositive tissues were detected in Group 3 and Group 4. In the histochemical examination, degenerative tubule structure and separation of epithelial cells were observed in I/R rats, while partially healed testicular tissue was detected in Group 3 and Group 4.

In our study, we observed that osthole reduced oxidative damage, suppressed the inflammatory process, prevented apoptosis, and reduced cell damage. We think that with repeated doses, cellular damage would gradually decline.