Guhan Yangsheng Jing mitigates oxidative stress and ferroptosis to improve reproductive damage in Diabetic Male Rats.

ETHNOPHARMACOLOGICAL RELEVANCE: According to traditional Chinese medicine (TCM) theory, reproductive injury is primarily associated with kidney essence deficiency, with the kidney being the affected organ. Guhan Yangshengjing (GHYSJ) is a traditional Chinese patent medicine, its main ingredients of GHYSJ, such as Polygonatum sibiricum Redouté, Epimedium brevicornu Maxim., and Lycium barbarum L. are believed to have significant kidney-tonifying effects, which can improve reproductive damage.

AIM OF THE STUDY: This study aims to investigate the protective effects of GHYSJ, a traditional Chinese medicine formula, on diabetes-induced male reproductive damage.

METHODS

In this study, we employed LC/Q-TOF-MS to analyze the active components of GHYSJ. A diabetic rat model was established using a high-sugar high-fat (HSHF) diet in combination with streptozotocin (STZ). Sperm quality and motility were assessed, and testicular morphology and sex hormones (testosterone [T], follicle-stimulating hormone [FSH], and luteinizing hormone [LH]) levels were examined to evaluate the impact of diabetes on reproductive function. Transcriptomic analysis was conducted to elucidate the potential mechanisms underlying GHYSJ's protective effects against diabetes-induced reproductive damage. Additionally, we used ELISA, immunofluorescence, transmission electron microscopy (TEM), immunohistochemistry, and Western blot to measure the expression levels of oxidative stress and ferroptosis-related markers, including oxygen species (ROS), superoxide dismutase (SOD), malondialdehyde (MDA), glutathione (GSH), lipid peroxidation (LPO), ferrous ion (Fe2+), nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), glutathione peroxidase 4 (GPX4), and cystine/glutamate antiporter (xCT).

RESULTS

Diabetic rats induced by a HSHF diet combined with STZ exhibited decreased sperm count, reduced sperm motility, and disrupted sex hormone secretion. GHYSJ intervention significantly reduced ROS levels and MDA accumulation in testicular tissue while enhancing SOD activity, thereby effectively alleviating oxidative damage. Additionally, GHYSJ modulated the Nrf2/HO-1 signaling pathway associated with oxidative stress, restoring testicular antioxidant capacity. This was evidenced by increased GSH levels, upregulated expression of antioxidant proteins (GPX4, xCT), decreased Fe2+ content, and reduced LPO levels. These effects collectively inhibited ferroptosis in testicular tissue of diabetic rats, leading to improved reproductive function.

CONCLUSIONS

Our findings demonstrate that GHYSJ exerts significant protective effects against diabetes-induced male reproductive damage by modulating oxidative stress and ferroptosis pathways. GHYSJ's ability to enhance antioxidant defenses and inhibit ferroptosis highlights its potential as a therapeutic agent for managing reproductive dysfunction in diabetic males.