The multigenerational effects of preconceptional administration of Chinese herbal medicine on male offspring in a murine model of polycystic ovary syndrome

Preconceptionally administered Bu-Shen-Tian-Jing Formula (BSTJF), a patented Chinese herbal medicine, has been proven to improve fertility in women with polycystic ovary syndrome (PCOS) and the pubertal neurobehavioral development of female offspring. This study aims to explore its multigenerational effects on the male offspring of the PCOS murine model. A PCOS murine model was established via prenatal anti-Müllerian hormone exposure. BSTJF was administered pre-conception, and its effects were evaluated in zero-generation (F0) mice and their first- and second-generation (F1/F2) male offspring. Reproduction, glucolipid metabolism, and the intestinal milieu phenotypes of male offspring were analysed via histological staining, biochemical analysis, 16 S rRNA sequencing, liquid chromatography-mass spectrometry untargeted metabolomics, and microbiota-metabolite correlation analysis. Preconceptionally administered BSTJF alleviated PCOS-related reproductive phenotypes in F0 mice. In both F1- and F2-PCOS male offspring, BSTJF reduced hyperandrogenemia, Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) indices, and hepatic lipid droplets. Furthermore, BSTJF attenuated serum low-density lipoprotein cholesterol levels, renal lipid deposition, insulin levels, seminiferous tubule damage and intestinal barrier impairment in F1, and total bilirubin levels and subcutaneous adipocyte size in F2. Microbiota-metabolite correlation analysis showed that preconceptional BSTJF administration altered key microbial hubs and liver metabolite associations in F1 and F2 male offspring, particularly the negative correlation between Akkermansia and LysoPC. The preconceptional administration of BSTJF attenuated the abnormalities in F0-PCOS mice, and was associated with amelioration of reproductive abnormalities, glucolipid metabolic dysregulation, gut microbiota dysbiosis, and hepatic metabolomic disturbance in F1 and F2 male offspring, possibly via the gut-liver axis.