Combining metabolomics and network pharmacology to clarify the mechanism of Zuogui Wan in treatment of oligoasthenospermia.

Zuogui Wan (ZGW), a classic traditional prescription used for nourishing kidney yin, has demonstrated considerable efficacy in the treatment of oligoasthenospermia (OAS) in both clinical and laboratory studies. However, the specific mechanism by which ZGW ameliorates OAS remains unclear. In this study, we integrated metabolomics and network pharmacology to explore the potential mechanisms underlying the therapeutic effects of ZGW in OAS. Pharmacodynamic studies revealed that ZGW increased body and testicular weights in mice and improved semen parameters. Furthermore, the testicular structure of the OAS mice was also significantly improved. Non-targeted metabolomics analysis identified 14 metabolites that were significantly altered (VIP > 1.0, p < 0.05), which were mainly enriched in the arginine and proline metabolism. Network pharmacology revealed 94 intersecting targets, and the enriched pathways also included the arginine and proline metabolism. Network pharmacology combined with metabolomics analysis identified the arginine and proline metabolism as a core pathway, and ornithine aminotransferase (OAT) and arginase 1 (ARG1) as core targets. We experimentally demonstrated that in OAS mice, ZGW suppresses the expression of ARG1 while simultaneously promoting the expression of OAT. Molecular docking results indicate that the core component of ZGW binds effectively to these two key targets. This study is the first to combine metabolomics and network pharmacology to investigate how ZGW improves OAS in mice via the arginine and proline metabolism, offering a new approach to the treatment of OAS.