Evaluation of Efficacy of Icariin Against Heat Stress-Induced Spermatogenic Dysfunction in Testes of Dogs

Baoan Li¹Liuwei Xie¹Mingqiang Song¹Xiumin Zhang²Chuanguo Yan¹Wanpeng Gao¹Wenxing Wang²Yang Yang^3*

• ¹College of Police Dog Technology of Criminal Investigation Police University of China, Shenyang, China
• ²Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha, China
• ³The Second Affiliated Hospital of Shenyang Medical College, Shenyang, China

The study examines the detrimental effects of high-temperature environments on dog testicular function, the challenges it poses to dog reproductive health, and explores the potential of Epimedii Folium, particularly its active component icariin, in mitigating these effects and enhancing testicular function.A completely randomized single-factor design was employed, involving 24 adult male Beagle dogs (9.82 ± 0.73 kg) randomly assigned to four treatment groups, with six dogs per group. The treatments comprised a negative control group (Control), a positive control group subjected to testicular heat stress (Model), and two groups administered 0.5 g/kg (Icariin-L) and 1.0 g/kg (Icariin-H) of icariin daily, respectively. All groups, except the negative control, underwent a testicular heat stress model to induce damage and assess icariin's effects on sperm quality, testicular function, hormone levels, protein expression, and testicular histology.The icariin supplementation improved sperm quality under heat stress, as evidenced by an increase in total count and motility, and a reduction in abnormality rates (P<0.01). It restored ATPase activity (Na⁺-K⁺-ATPase, Mg²⁺-ATPase, Ca²⁺-ATPase) and serum hormone levels (GnRH, LH, E2) (P<0.01). Western blot analysis revealed icariin upregulated steroidogenic proteins (STAR, 17βHSD, CYP450) and tight junction protein ZO-1 (P<0.01), while and suppressed BAX expression (a key regulator of mitochondrial apoptosis) and enhanced BCL-2 expression (a major anti-apoptotic factor in the BCL-2 family) (P<0.01). Histological assessments demonstrated icariin mitigated heat-induced seminiferous tubule damage, epithelial thinning, and spermatogonia degeneration. Molecular docking confirmed strong binding affinity between icariin and 17βHSD via hydrogen bonding and hydrophobic interactions.These findings suggest that icariin appears to alleviate testicular heat stress potentially by canine 删除[明强 宋]: canine 删除[明强 宋]: canines 删除[明强 宋]: canines 删除[明强 宋]:modulating testosterone synthesis, enhancing ATPase function, restoring blood-testis barrier integrity, and inhibiting apoptosis. The dose-dependent efficacy (1.0 g/kg > 0.5 g/kg) suggests its potential as a possible therapeutic agent for dog reproductive health in high-temperature environments.