YiJing Powder activates the HO-1/NRF2/KEAP1 antioxidant axis to remodel lipid metabolism and ameliorate oligoasthenospermia.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: YiJing Powder (YJP) is a traditional Chinese herbal formula clinically prescribed for kidney-yang deficiency-associated oligoasthenospermia (OA), a prevalent cause of male infertility characterized by diminished sperm count and motility. Despite its empirical efficacy in restoring spermatogenic function, the pharmacodynamic basis of YJP remains poorly understood. AIM OF THE STUDY: This study integrates phytochemical analysis and experimental validation to systematically decipher the mechanism by which YJP ameliorates OA, with a focus on its regulation of the HO-1/NRF2/KEAP1 antioxidant axis and lipid metabolic reprogramming. MATERIALS AND METHODS: YJP components were determined by HPLC. An OA rat model was established using ornidazole-induced induction. Therapeutic effects of YJP were evaluated via H&E staining, ELISA, and biochemical assays. Non-targeted lipid metabolomics was used to identify dysregulated metabolic pathways in testicular tissues. WB and RT-qPCR were performed to validate key proteins and genes, revealing YJP's regulation via the HO-1/NRF2/KEAP1 axis. RESULTS: HPLC identified nine bioactive compounds in YJP, including protocatechuic acid, rehmannioside D, ferulic acid, etc. YJP treatment improved sperm motility and density and enhanced antioxidant capacity, as shown by increased SOD, GSH, and NO levels. Histopathology indicated recovery of testicular and epididymal morphology, with preserved ultrastructural integrity. Lipidomics showed YJP normalized Cer, PE, and CL levels by modulating glycerophospholipid and sphingolipid metabolism. Mechanistically, YJP activated the HO-1/NRF2/KEAP1 pathway. CONCLUSION: YJP ameliorates OA through multi-target mechanisms, including redox homeostasis restoration via HO-1/NRF2/KEAP1 and lipid metabolism regulation by correcting Cer/PE/CL imbalance, which is essential for sperm membrane integrity.