Abstract: 【Objective】 This study aimed to investigate the effects of mulberry leaf powder on production performance and cecal microbial community structure of laying hens， providing theoretical reference for developing and utilizing mulberry leaf powder， improving the health of laying hens， and enhancing egg quality. 【Method】 A total of 120 Hyline brown laying hens were randomly divided into four groups： hens in the control group （CK） were fed basal diets， and hens in the experimental groups were fed basal diets supplemented with 3% mulberry leaf powder （3% ML）， 6% mulberry leaf powder （6% ML）， and 9% mulberry leaf powder （9% ML）. Egg production rate， average daily feed intake， average egg weight， and feed-egg ratio were calculated， while egg shape index， eggshell thickness， albumen height， and yolk color were measured. The egg shape index and Haugh unit were calculated. Hematoxylin-eosin （HE） staining was used to observe cecal tissues. The serum antioxidant and immune indicators were measured using assay kits. The diversity and composition of the cecal microbiota were analyzed through 16S rRNA gene high-throughput sequencing. 【Result】 With increasing dietary mulberry leaf powder supplementation， egg production rate of laying hens showed a decreasing trend， with the 9% ML group showing a significantly lower egg production rate than the CK （P<0.05， the same below）； the 6% ML group had the highest albumen height and Haugh units. Cecal villus length was longer in both the 3% ML and 6% ML groups than in the CK， while the cecal villus length was significantly shorter in the 9% ML group than in the CK； the cecal villus width was the largest in the 3% ML group； and the crypt depth was significantly lower in the 9% ML group than in the CK， 3% ML， and 6% ML groups. Total antioxidant capacity of serum （serum T-AOC）， superoxide dismutase （SOD） activity， and immunoglobulin G （IgG） content were significantly higher in the 3% ML group than in the CK， while tumor necrosisfactor α （TNF-α） content was significantly lower in the 3% ML group. Correlation analysis showed that egg production rate was significantly negatively correlated with the relative abundance of Lactobacillus； villus length was positively correlated with the relative abundance of Lactobacillus， Akkermansia， and Desulfovibrio， but negatively correlated with the relative abundance of uncultured_bacterium_f_Muribaculaceae； serum T-AOC was significantly positively correlated with the relative abundance of Lactobacillus， Akkermansia， and Desulfovibrio， but significantly negatively correlated with the relative abundance of uncultured_bacterium_f_Muribaculaceae. 【Conclusion】 Effects of supplementing mulberry leaf powder to chicken diets on production performance and gut health of laying hens are obviously dose-dependent. Supplementing an appropriate amount （3%） of mulberry leaf powder can regulate the gut microbiota structure of laying hens to promote the proliferation of probiotics and inhibit the growth of harmful bacteria， and increase antioxidant enzyme activity and immunoglobulin secretion levels， ultimately improving the health and production performance of laying hens.