Salidroside Represses the Viability of Mouse Sertoli Cell Line (TM4 Cells) via Repression of Ldha-Dependent Glycolysis

Salidroside is a bioactive compound extracted from the traditional Chinese medicinal plant Rhodiola rosea that exerts differential regulatory effects on cell viability depending on cell type and physiological state. Sertoli cells are important testicular somatic cells that play multiple roles in nourishing, protecting, and supporting spermatogenic cells. Currently, the regulatory effects and underlying mechanisms of salidroside on Sertoli cells remain unclear. This study investigated the effects of different concentrations of salidroside on the viability and regulatory mechanisms of TM4 cells (mouse Sertoli cell line). The results showed that within the range of 25–600 μM, cell viability decreased with increasing salidroside concentration. Treatment with 600 μM salidroside inhibited proliferation, promoted apoptosis, and suppressed the expression of key genes related to Sertoli cell function. Transcriptome sequencing revealed that several glycolysis-related genes, including Ldha, Pfkl, Eno3, and Eno1b, were significantly downregulated in salidroside-treated cells, accompanied by marked reductions in key glycolytic metabolites, including fructose-1,6-diphosphate (FDP), pyruvate, lactic acid, and ATP. Notably, overexpression of lactate dehydrogenase A (Ldha) rescued cell viability and proliferation, suppressed apoptosis, elevated both intracellular and extracellular lactate and ATP levels, and partially restored functional gene expression in 600 μM salidroside-treated cells. Overall, this study provides hitherto undocumented evidence of the regulatory effects and mechanisms of salidroside on TM4 cells, identifying the Ldha–glycolytic axis as an in vitro cellular target of high-dose salidroside in Sertoli cells, which may inform future in vivo reproductive safety profiling pending pharmacokinetic validation.