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Background    SMAD family proteins (SMADs) are crucial transcription factors downstream of transforming growth factor beta (TGF-ß)/SMAD signaling pathways that have been reported to play a pivotal role in mammalian reproduction. However, the role of SMAD family member 8 (SMAD8, also known as SMAD9), a member of the SMAD family, in mammalian reproduction remains unclear.Methods    We employed RNA interference techniques to knock down  Smad8  expression in mouse granulosa cells (GCs) to investigate the effects of  Smad8  on GC growth and steroidogenesis.Results    Our findings revealed a significant decrease in the proliferative capacity and a substantial increase in the apoptosis rate of GCs after transfection with  Smad8 -siRNA for 48 h. Subsequent hormone assays demonstrated a significant decrease in estradiol (E 2 ) levels, whereas progesterone (P 4 ) remained unchanged. Further mechanistic analysis showed that the mRNA expression of proliferating cell nuclear antigen ( Pcna ),  Cyclin D2 , cell cycle-dependent kinase 4 ( Cdk4 ), B-cell lymphoma-2 ( Bcl-2 ), estrogen receptor ( Er ), luteinizing hormone receptor ( Lhr ) and cytochrome P450 family 19 subfamily A member 1 ( Cyp19a1 ) significantly decreased. Conversely, the mRNA of cysteine aspartate proteinase 3 ( Caspase 3 ) significantly increased, wheras  Bcl2 -associated X ( Bax ), follicle-stimulating hormone receptor ( Fshr ) and cytochrome P450 family 11 subfamily A member 1 ( Cyp11a1 ) remained unchanged compared to the controls.Conclusion    This study indicates that  Smad8  knockdown inhibits cell proliferation, promotes apoptosis, reduces  Er  and  Lhr  transcription, and decreases E 2  production in mouse GCs. These findings suggest that  Smad8  may serve as a novel genetic marker for mammalian reproduction.

Smad8 is involvement in follicular development via the regulation of granulosa cell growth and steroidogenesis in mice