Life sciences · 2026
TRPM8 activation by sustained-release menthol mitigates exertional heatstroke-induced testicular injury and sperm ultrastructural damage in rats.
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Ruan Ting, Chao Chien-Ming, Shen Chung-Min, Hsieh Kun-Lin, Chen Tzu-Hao, Tang Ling-Yu, Tan Tian-Hoe, Lin Yu-Jung, Chang Ching-Ping
Аннотация
AIMS: Exertional heatstroke (EHS) can induce marked scrotal hyperthermia and impair male reproductive integrity. This study tested whether sustained-release menthol protects against EHS-induced testicular and genitourinary injury via transient receptor potential melastatin 8 (TRPM8) activation.
MATERIALS AND METHODS: Adult male Wistar rats received sustained-release menthol gel (30 mg·kg-1, subcutaneously) or vehicle for 2 weeks before EHS induction (36 °C, 50% relative humidity; approximately 52 min of treadmill running). Some rats received AMTB, a selective TRPM8 antagonist. Scrotal temperature change (ΔTscrotal) and circulating hormones were measured, and genitourinary tissues were examined on Day 3 after EHS. Blood-testis barrier proteins (claudin-11 and ZO-1), TRPM8, and apoptosis of Sertoli and Leydig cells were quantified by four-colour immunofluorescence, and sperm ultrastructure was assessed by scanning and transmission electron microscopy.
KEY FINDINGS: Vehicle-treated EHS rats showed marked scrotal hyperthermia (ΔTscrotal: 6.5 °C vs 0.75 °C in controls), reduced testosterone with elevated luteinizing hormone, sperm dysmorphology, and multisite genitourinary injury. EHS reduced claudin-11/ZO-1 and TRPM8 in seminiferous tubules and increased Sertoli/Leydig cell apoptosis. Electron microscopy revealed sperm head deformation, head-neck discontinuity, mitochondrial sheath disruption in the midpiece, and fissures/perforations along the principal piece. Menthol pretreatment attenuated scrotal hyperthermia (ΔTscrotal: 2.6 °C), improved testosterone, reduced Sertoli and Leydig cell apoptosis, and alleviated histological and ultrastructural injury. These benefits were blunted by AMTB, supporting TRPM8-dependent protection.
SIGNIFICANCE: Sustained-release menthol mitigates EHS-induced male reproductive and genitourinary injury in rats, at least partly through TRPM8 activation. TRPM8 may represent a potential prophylactic target for reducing heat-related reproductive damage.
MATERIALS AND METHODS: Adult male Wistar rats received sustained-release menthol gel (30 mg·kg-1, subcutaneously) or vehicle for 2 weeks before EHS induction (36 °C, 50% relative humidity; approximately 52 min of treadmill running). Some rats received AMTB, a selective TRPM8 antagonist. Scrotal temperature change (ΔTscrotal) and circulating hormones were measured, and genitourinary tissues were examined on Day 3 after EHS. Blood-testis barrier proteins (claudin-11 and ZO-1), TRPM8, and apoptosis of Sertoli and Leydig cells were quantified by four-colour immunofluorescence, and sperm ultrastructure was assessed by scanning and transmission electron microscopy.
KEY FINDINGS: Vehicle-treated EHS rats showed marked scrotal hyperthermia (ΔTscrotal: 6.5 °C vs 0.75 °C in controls), reduced testosterone with elevated luteinizing hormone, sperm dysmorphology, and multisite genitourinary injury. EHS reduced claudin-11/ZO-1 and TRPM8 in seminiferous tubules and increased Sertoli/Leydig cell apoptosis. Electron microscopy revealed sperm head deformation, head-neck discontinuity, mitochondrial sheath disruption in the midpiece, and fissures/perforations along the principal piece. Menthol pretreatment attenuated scrotal hyperthermia (ΔTscrotal: 2.6 °C), improved testosterone, reduced Sertoli and Leydig cell apoptosis, and alleviated histological and ultrastructural injury. These benefits were blunted by AMTB, supporting TRPM8-dependent protection.
SIGNIFICANCE: Sustained-release menthol mitigates EHS-induced male reproductive and genitourinary injury in rats, at least partly through TRPM8 activation. TRPM8 may represent a potential prophylactic target for reducing heat-related reproductive damage.