Abstract: This study was conducted to investigate the metabolites, metabolic hormones and reproductive hormones concentrations in differentsized follicles in Hu sheep during the luteal phase. 11 multiparity Hu sheep with 40 kg in body weight were used. On the 12th day after estrus synchronization, all ewes were slaughtered and follicular fluid was collected according to differentsized follicles. The effects of follicle size on serum ammonia, NEFA, urea, insulin and progesterone concentrations were not significant. Compared with follicles ≤2.5 mm in diameter, the follicular fluid glucose and estradiol concentrations in follicles >2.5 mm in diameter were increased (P<0.05), and the testosterone and glucagon concentrations and LDH activity were decreased (P<0.05). The estradiol concentration in follicular fluid was positively correlated with glucose (P<0.05) and progesterone (P=0.051) concentration and negatively correlated with LDH (P<0.01), glucagon (P<0.05) and testosterone (P<0.01) concentration. In conclusion, follicle development was coregulated by intrafollicular metabolites and hormones.