Abstract

Simple SummaryExosomes in the follicular fluid can carry and transfer regulatory molecules to recipient cells, thus influencing their biological functions. In this study, after being treated with yak follicular fluid for 24 h, most yak cumulus cells took up the exosomes, activated autophagy, and increased their secretion of 2-hydroxyestrodiol. Conversely, the inhibition of autophagy with the chemical reagent 3-methyladenine blocked these effects, suggesting that autophagy has an important role in 2-hydroxyestrodiol secretion in the yak cumulus cells. The results of rapamycin treatment of the yak cumulus cells showed a similar effect on yak follicular fluid exosomes as there was an increase in 2-hydroxyestrodiol secretion due to the activation of autophagy. The results suggest that autophagy is activated by yak follicular fluid exosomes and this increases 2-hydroxyestrodiol secretion in the yak cumulus cells.AbstractExosomes in the follicular fluid can carry and transfer regulatory molecules to recipient cells, thus influencing their biological functions. However, the specific effects of yak follicular fluid exosomes on 2-hydroxyestrodiol (2-OHE2) secretion remain unknown. Here, we investigated whether yak follicular fluid exosomes can increase 2-OHE2secretion through the activation of autophagy in cumulus cells (YCCs). In vitro cultured YCCs were treated with yak follicular fluid exosomes for 6, 12, and 24 h. The effects of yak follicular fluid exosomes on autophagy and 2-OHE2secretion were evaluated through real-time quantitative fluorescence PCR (RT-qPCR), Western blotting (WB), transfected with RFP-GFP-LC3, immunohistochemistry, and ELISA. To further investigate whether 2-OHE2secretion was related to autophagy, YCCs were administered with yak follicular fluid exosomes, 3-methyladenine (3-MA), and rapamycin (RAPA). The results revealed that treatment with yak follicular fluid exosomes activated autophagy in YCCs and increased 2-OHE2secretion. Conversely, the inhi