Leech therapy (Hirudo medicinalis) attenuates testicular damages induced by testicular ischemia/reperfusion in an animal model.

BACKGROUND: Testicular torsion/detorsion triggers tissue ischemia/reperfusion, leading to reactive oxygen species overgeneration and apoptosis. The saliva of leeches is full of anti-inflammatory, anticoagulants, antioxidants, and antimicrobial agents. Therefore, this study aimed to assess the protective mechanism of leech therapy on testicular ischemia/reperfusion damage. METHODS: 18 adult male rats were randomly divided into three groups: 1-Sham-operated group (SO). 2-Torsion/detorsion (T.D) group: two hours of testicular torsion with two hours of testicular detorsion was performed. 3-Torsion/detorsion + Leech therapy (TDL) group. Sperm parameters (motility, vitality, morphology, and concentration), oxidative stress biomarkers (MDA, CAT, GPx, and TAC), histopathological factors (Mean seminiferous tubular diameter, Germinal epithelial cell thickness, Testicular capsule thickness, Johnson's score, and Cosentino's score), and immunohistochemical markers for apoptosis detection (Bax, Bcl-2, and Caspase-3) were measured. RESULTS: There was a significant difference for all sperm parameters in the T. D group compared to the sham group. Leech therapy significantly increased progressive motility and normal morphology and reduced non-progressive motility. In the TDL group, MDA concentration significantly reduced, and levels of GPx, TAC, and CAT remarkably increased. All evaluated histopathological parameters in the TDL group significantly increased compared to the T. D group except for the testicular capsule thickness. T. D notably increased the expression of Bax and Caspase-3, while the treatment group slowed the rate of apoptosis compared to the control group. Bcl-2 expression in the T. D group was significantly lower than that in the sham group. Leech therapy increased the Bcl-2 expression. CONCLUSION: Leech therapy attenuates damages to testicular tissue following torsion/detorsion due to its antioxidant, anti-inflammatory, and anti-apoptotic effects. Hence, it can be considered as an effective remedy for testicular ischemia/reperfusion.

Salvia miltiorrhiza hydroalcoholic extract inhibits postoperative peritoneal adhesions in rats.

BACKGROUND: One of the most prevalent postoperative complications is believed to be intra-abdominal peritoneal adhesions, which is followed by several complications. Several adhesion prevention products have been examined, yet none of them were found to be completely effective. The current research is conducted to evaluate the beneficial effects of Salvia miltiorrhiza hydroalcoholic extract in inhibiting postoperative peritoneal adhesions in rats. METHODS: Forty rats were randomly classified into five equal groups (n = 8): 1) the normal group did not undergo surgical operations, 2) the control group in which the adhesion was induced, and which did not receive any treatment, 3) distilled water group that received distilled water, and 4,5) treatment groups treated with 1 and 5% of Salvia miltiorrhiza hydroalcoholic extract. The rats were euthanized 14 days following the surgery and the macroscopic score, the microscopic score of granulomatous inflammation and granulation tissue formation, IHC markers (vimentin, CD31, IL-1ß, COX-2, and iNOS), and oxidative stress biomarkers (MDA, GPx, CAT, and TAC) were assessed in the experimental groups of the study. RESULTS: The difference between the control group and other groups for the adhesions macroscopic score, microscopic score, IHC markers, and oxidative stress biomarkers was significant (p < 0.05). Distilled water had no protective effect on the formation of peritoneal adhesions. Salvia miltiorrhiza treatment in two different doses significantly reduced macroscopic and microscopic scores, MDA concentration, Vimentin, IL-1ß, COX-2, and iNOS compared to the control group (p < 0.05). The levels of GPx, CAT, and TAC in the treatment groups increased significantly compared with the control group (p < 0.05). Our findings revealed that a higher dose of Salvia miltiorrhiza was more effective in reducing peritoneal adhesions, proinflammatory and mesenchymal cell markers, and oxidative stress. CONCLUSIONS: Salvia miltiorrhiza extract, owing to its strong antioxidant and anti-inflammatory properties, could effectively reduce peritoneal adhesions. Therefore, Salvia miltiorrhiza is recommended to be used as an effective anti-peritoneal post-operative adhesive agent.

Co-administration of Salvia miltiorrhiza and verapamil inhibits detrimental effects of torsion/detorsion on testicular tissue in rats.

Testicular torsion/detorsion is one of the important emergencies that requires fast surgical intervention. This study aimed to investigate the effects of Salvia miltiorrhiza hydroalcoholic extract combined with verapamil on testicular ischaemia/reperfusion damage in Wistar albino rats. All animals were distributed in 3 groups (n = 8), including the sham-operated group, torsion/detorsion (TD) group and torsion/detorsion + pretreatment with 200 mg/kg Salvia miltiorrhiza extract combined with 0.3 mg/kg verapamil (SMV) group. Oxidative stress biomarkers (MDA, GPx, CAT and TAC) both in plasma and testicular tissue, sperm parameters (motility, vitality, concentration and morphology) and histopathological parameters (MSTD, GECT, Johnson's score, Cosentino's score and testicular cell thickness) were assessed in all groups. Ischaemia/reperfusion significantly increased MDA and decreased GPx, CAT and TAC levels (p < .05). Pretreatment with SMV significantly increased GPx, CAT and TAC levels (p < .05). SMV group increased progressive sperm motility and vitality and reduced non-progressive motility of spermatozoon (p < .05). Testicular torsion significantly decreased all histopathological parameters compared to the sham group (p < .05). SMV pretreatment remarkably increased MSTD, GECT and Cosentino's score in comparison with the TD group (p < .05). A combination of Salvia miltiorrhiza with verapamil could reduce damages triggered by testicular torsion detorsion and improve sperm functionality parameters and oxidative stress defence systems.

Nitroglycerin ameliorates sperm parameters, oxidative stress and testicular injury following by testicular torsion/detorsion in male rats.

PURPOSE: This study was designed to determine the probable protection mechanisms of nitroglycerin, a widely used medication for treatment of heart failure and angina, in amelioration of testicular ischemia/reperfusion damage. METHODS: 24 adult male rats were randomly divided into three equal groups; with eight rats in each group: Group 1 (Sham) was sham-operated. Group 2 (T_D): 2 h testicular torsion was induced, afterward detorsion was performed and maintained for 2 h. Group 3 (NG): Nitroglycerin was administered immediately after detorsion. Sperm quality parameters such as viability, motility, morphology, and concentration, levels of antioxidant enzymes (glutathione peroxidase (GPX), catalase (CAT), and total antioxidant capacity (TAC)), and amount of malondialdehyde (MDA) in the blood plasma were examined in each group, thereafter histopathological parameters including germinal epithelial cell thickness (GECT), mean seminiferous tubular diameter (MSTD), Johnson's score and Cosentino's score were assessed. RESULTS: Testicular T_D significantly reduced sperm viability, motility, and normal morphology, whereas the NG administration remarkably increased the percentage of live, motile, and normal spermatozoa (p < 0.05). Levels of GPx, CAT, and TAC significantly reduced and the MDA level significantly increased in the T_D group in comparison to the sham group (p < 0.05). The NG treated group demonstrated significantly reduced MDA concentrations as well as elevated levels of GPx and CAT compared to the T_D group (p < 0.05). Induction of testicular torsion significantly reduced Johnson's score, GESCT (µm), and MSTD (µm), and remarkably increased the Cosentino's score (P < 0.05), while NG injection significantly increased Johnson's score, GESCT (µm), and MSTD (µm) and reduced the Cosentino's score (P < 0.05). CONCLUSION: According to the findings in this research, nitroglycerin was able to protect the testicular tissue from ischemia-reperfusion damage caused by induced torsion/detorsion.

Investigating the sperm parameters, oxidative stress and histopathological effects of salvia miltiorrhiza hydroalcoholic extract in the prevention of testicular ischemia reperfusion damage in rats.

AIMS: One of the most common urologic emergencies is spermatic cord torsion, which can damage testicular tissue and reduce fertility. Salvia miltiorrhiza (SM) hydroalcoholic extract possess high antioxidant properties, and its efficacy in ischemia-reperfusion (I/R) injury prevention has been demonstrated in cardiac, renal, and liver tissues. Therefore, the purpose of this study was to assess the protective mechanism of SM extract on testicular I/R damage. MAIN METHODS: 18 mature male Wistar albino rats were randomly divided into 3 groups; with six rats in each group: Group 1 (Sham) was sham-operated. Group 2 (T-D): torsion was performed, and after 2 hours (h) detorsion was done. Group 3 (SM): (200 mg kg-1) SM was intraperitoneally injected thirty minutes before detorsion. Then testicular and epididymal weight and size alterations, sperm parameters (motility, livability, concentration, and morphology), both plasma and testicular tissue levels of malondialdehyde (MDA), catalase (CAT), glutathione peroxidase (GPX), and total antioxidant capacity (TAC) were evaluated. Also, histopathological changes included mean seminiferous tubular diameter (MSTD), testicular capsule thickness (TCT), mean testicular biopsy scoring (MTBS), and germinal epithelial cell thickness (GECT) were examined. RESULTS: Testicular I/R significantly reduced sperm motility, viability, and normality, while SM extract administration remarkably increased sperm motility, and normality (P < 0.05). Induction of testicular T-D caused a significant increment in the level of MDA and notable decline in the levels of GPX, CAT, and TAC both in plasma and testis tissue, whereas administration of SM extract significantly decreased MDA level and increased GPX, CAT, and TAC levels in plasma and testicular tissue (P < 0.05). Histopathological parameters including MSTD, GECT, MTBS, and TCT were significantly lower in the T-D group, while pretreatment with SM extract remarkably increased MSTD, GECT, and MTBS amounts (P < 0.05). CONCLUSION: Since the SM extract increased the activity of antioxidant enzymes, improved sperm parameters and reduced the damage to testicular tissue, therefore, its use as a potent antioxidant in reducing testicular I/R damage is suggested.