Metformin Prevents Follicular Atresia in Aging Laying Chickens through Activation of PI3K/AKT and Calcium Signaling Pathways

Increased follicular atresia occurs with aging and results in reduced fecundity in laying chickens. Therefore, relieving follicular atresia of aging poultry is a crucial measure to maintain sustained high laying performance. As an antiaging agent, metformin was reported to play important roles in preventing aging in diverse Animals. In this study, the physiological state of the prehierarchical follicles in the peak-laying hens (D280) and aged hens (D580) was compared, followed with exploration for the possible capacity of metformin in delaying atresia of the prehierarchical follicles in the aged D580 hens. Results showed that the capacity of yolk deposition within follicles declined with aging, and the point of endoplasmic reticulum- (ER-) mitochondrion contact decreased in the ultrastructure of the follicular cells. Meanwhile, the expression of Apoptosis signaling genes was increased in the atretic small white follicles. Subsequently, the H₂O₂-induced follicular atresia model was established to evaluate the enhancing capacity of metformin on yolk deposition and inhibition of Apoptosis in the atretic small white follicles. Metformin inhibited Apoptosis through regulating cooperation of the mitochondrion-associated ER membranes and the Insulin (PI3K/Akt) signaling pathway. Furthermore, metformin regulated calcium ion homeostasis to relieve ER-stress and inhibited release of mitochondrion Apoptosis factors (BAD and Caspase). Additionally, metformin activated PI3K/Akt that suppressed activation of BAD (downstream of the Insulin signaling pathway) in the atretic follicles. Further, serum estrogen level and liver estrogen receptor-α expression were increased after dietary metformin supplementation in D580 hens. These results indicated that administration of dietary metformin activated the PI3K/Akt and calcium signaling pathway and enhanced yolk deposition to prevent chicken follicular atresia.