[Erjing Wan ameliorates busulfan-induced oligoasthenospermia in mice by inhibiting p38 MAPK/NF-κB pathway]

Oligoasthenospermia is a major cause of male infertility. According to the TCM theory that the kidney stores essence and governs reproduction, this study investigated the therapeutic effects and mechanisms of the classic formula Erjing Wan(EJW) on busulfan(Bu)-induced oligoasthenospermia in mice. A mouse model of oligoasthenospermia was established by Bu injection, and the water decoction(EJW-W), alcohol extract(EJW-E), and polysaccharide fraction(EJW-P) of EJW were systematically prepared, in which the chemical constituents were analyzed by UPLC-MS/MS. In vivo pharmacodynamic evaluation showed that compared with the model group, all EJW interventions significantly increased the sperm count and motility and restored the serum levels of testosterone(T) and follicle-stimulating hormone(FSH). Additionally, testicular oxidative stress was effectively ameliorated, as indicated by enhanced activities of superoxide dismutase(SOD) and glutathione(GSH), along with reduced malondialdehyde(MDA) content. Histopathological examination revealed significant alleviation of seminiferous tubule vacuolization and structural atrophy, with improved orderly arrangement of spermatogenic cells at various stages. The further molecular mechanism study demonstrated that the therapeutic effects were closely associated with the suppression of the p38 mitogen-activated protein kinase(MAPK)/nuclear factor-kappa B(NF-κB) signaling pathway over-activation, manifested as significantly downregulated expression of phosphorylated p38 MAPK(p-p38 MAPK), phosphorylated NF-κB p65(p-NF-κB p65), and downstream inflammatory cytokines interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α). Moreover, the expression of synaptonemal complex protein 3(SYCP3), a marker of spermatogenic cells, and cholesterol side-chain cleavage enzyme(CYP11A1~+), a key enzyme in testosterone synthesis, was effectively up-regulated. Notably, EJW-E and EJW-P exhibited particularly remarkable effects, with EJW-P showing superior potential in improving the testicular microenvironment. In conclusion, EJW extracts ameliorate Bu-induced oligoasthenospermia by inhibiting the p38 MAPK/NF-κB signaling pathway and mitigating testicular oxidative stress and inflammatory responses. The active material basis is likely associated with flavonoids and steroidal saponins in the alcohol extract, as well as polysaccharides.