Chronic exposure to liquid crystal monomer EBCN at environmentally relevant concentrations induces testicular dysfunction via the gut-testis axis.

4-Cyano-4'-ethoxybiphenyl (EBCN) is a representative cyano liquid crystal monomer (LCM). While prior studies have documented the widespread occurrence of LCMs in diverse environmental and biological samples, research on their reproductive effects in vivo remains limited. This study employed 35-day and 70-day exposure models in mice to assess the short-term and long-term effects of environmentally relevant concentrations of EBCN on testicular health. Our findings indicate that EBCN exposure, irrespective of duration, had minimal impact on body weight, testis weight, and testicular organ coefficient. However, it induced dose-dependent reductions in seminiferous tubule area, sperm count, accompanied by decreases in Leydig cells and spermatogenic cells, along with disruptions in sex hormone levels. Moreover, EBCN exposure led to the upregulation of inflammatory factors in serum, partially attributable to the activation of necroptosis-related pathways. Additionally, 16S rRNA sequencing and metabolomic analysis revealed a decline in gut microbiome diversity and a decrease in anti-inflammatory metabolites, specifically L-carnosine, in the intestine, potentially contributing to the observed testicular toxicity. Supplementation with exogenous L-carnosine mitigated EBCN-induced testicular dysfunction by inhibiting the expression of necroptosis-related genes. In conclusion, our study suggests that prolonged EBCN exposure at environmentally relevant concentrations adversely impacts testicular function via the gut-testis axis.