Tuominjian decoction reduces the proportion of Tfh cells and B-cells in immune infertility male mice by regulating the seminal plasma levels of IL-1 and IL-6.

Abstract

BACKGROUND: Male immunological infertility (MII), driven by anti-sperm antibody (AsAb), is a complex autoimmune disorder. The herbal formulation TuominJian (TMJ) has shown clinical efficacy against MII, but its immunomodulatory mechanisms involving Tfh cells, B cells, and cytokines remain unclear. PURPOSE: This study aimed to elucidate the immunoregulatory effects and underlying mechanism of TMJ in MII mice. METHODS: Active compounds in TMJ were identified using UPLC-Q/TOF-MS. An MII mouse model was established using sperm immunization. After verifying the model, testicular and epididymal tissue integrity, sperm parameters, and blood-testis barrier function were assessed via H&E staining, TEM, CASA, and western blotting. Serum AsAb, cytokines, and immune cell populations were analyzed by ELISA and flow cytometry. RESULTS: Thirteen active compounds were identified in TMJ. TMJ treatment significantly reduced testicular cell apoptosis and decreased serum AsAb and proinflammatory cytokines (IL-1, IL-6). It also restored testicular and epididymal tissue architecture, improved sperm concentration and motility, and enhanced blood-testis barrier integrity by upregulating occludin. Furthermore, TMJ downregulated Tfh and B cell levels. CONCLUSION: TMJ mitigates MII in mice by promoting structural recovery of reproductive tissues, reinforcing the blood-testis barrier, and modulating immune responses through suppression of Tfh cells, B cells, and proinflammatory cytokines. These findings highlight immunoregulation as a promising therapeutic strategy for MII.