Autophagic and apoptotic proteins in goat corpus luteum and the effect of Adiponectin/AdipoRon on luteal cell autophagy and apoptosis

Abstract

The most abundant adipokine Adiponectin (APN) is present in ovaries. AdipoRon is a small molecule oral APN receptor agonist that binds and activates APN receptors. However, the function of APN/AdipoRon in regulation of luteal cell processes has not been elucidated. To investigate autophagic and apoptotic proteins in goat CLs and effects of APN/AdipoRon on goat luteal autophagy and apoptosis, goat CLs were collected during the early, mid and late luteal stages of the estrous cycle to evaluate autophagic and apoptotic protein patterns. LC3B, Beclin 1, Caspase-3 and Bax/Bcl-2 as well as p-AMPK were differentially abundant at different stages of CL development. All these proteins were primarily localized in large and small luteal steroidogenic cells. Then, isolated luteal steroidogenic cells were evaluated to ascertain the functions and mechanism of APN/AdipoRon in luteal autophagy and apoptosis. Treatment with AdipoRon (25 and 50 μM) and APN (1 μg/mL) for 48 h resulted in a decrease in cell viability and P4 level, increased autophagic and apoptotic proteins. Treatment with AdipoRon (25 μM) led to rapid and transient p-AMPK activation, with p-AMPK elevated at 30 min to 1 h with there being a return to a basal concentration at 2 h post-treatment. Moreover, treatment with AdipoRon led to an increase in autophagy by activating AMPK, which was markedly reduced with treatment with an AMPK inhibitor Compound C and siAMPK, however, abundances of apoptotic proteins were not affected by these treatments. In conclusion, autophagy and apoptosis are involved in the structural regression of goat CL. APN/AdipoRon led to a lesser cell viability and P4 concentration, and activated autophagy through induction of the AMPK while there was induction of apoptosis through an AMPK - independent pathway in goat luteal cells.