Intestinal integrity, digestive enzyme activity, nutrient utilization, and egg quality of laying hens at different ages were evaluated and compared in this study. A total of 192 Hy-line Brown laying hens at 195-d-old (D195 group), 340-d-old (D340 group), and 525-d-old (D525 group) were allocated into one of 3 groups in accordance with their ages. Each group had 8 replicates of 8 birds each, and all birds were fed a maize-soybean meal basal diet for a 2-wk experiment. Compared with the D195 group, intestinal villus height and ratio of villus height to crypt depth, as well as serum D-lactate content increased in the D525 group (
P
< 0.05). The sucrase and maltase activities in the jejunal mucosa, amylase activity in the pancreas, and trypsin activity in the jejunal chyme of 525-d-old hens were lower than their 195-d-old counterparts (
P
< 0.05). In addition, there was a decline of trypsin and lipase activities in the ileal chyme of hens from D525 group in comparison with D195 or D340 group (
P
< 0.05). Apparent retention of dry matter and crude protein of birds in D340 and D525 group decreased when compared with the D195 group (
P
< 0.05). Moreover, birds in the D525 group exhibited a lower level of ether extract retention, and higher contents of several excreted amino acids than those in the D195 group (
P
< 0.05). Compared with the D195 group, eggs harvested from D525 group exhibited lower albumen height, eggshell strength and thickness, and a higher egg weight (
P
< 0.05). In conclusion, increased intestinal permeability (higher serum D-lactate content), compromised digestive function (lower digestive enzyme activities and apparent nutrient retention, and higher concentrations of excreted amino acids), and poor egg quality (lower albumen height, eggshell strength, and thickness) were observed with increasing age in the laying hens.