BACKGROUND Stevioside (STE) is a widely used sweetener. Despite the fact that chickens are insensitive to sweetness, dietary STE supplementation could increase the feed intake of broiler chickens. Stevioside might regulate the feeding behavior through functional mechanisms other than its high‐potency sweetness. The present study was aimed to elucidate the potential sweetness‐independent mechanism of an STE‐induced orexigenic effect using the broiler chicken and considering the hypothalamic transcriptome profile and gut microbiome. RESULTS The analysis of RNA‐Seq identified 398 differently expressed genes (160 up‐regulated and 238 down‐regulated) in the hypothalamus of the STE‐supplemented group compared with the control group. Cluster analysis revealed several appetite‐related genes were differentially expressed, including NPY, NPY5R, TSHB, NMU, TPH2, and DDC. The analysis of 16S rRNA sequencing data also indicated that dietary STE supplementation increased the relative abundance of Lactobacillales, Bacilli, Lactobacillus, and Lactobacillaceae. Meanwhile, the proportion of Ruminococcaceae, Lachnospiraceae, Clostridia, and Clostridiales was decreased after dietary supplementation with STE. CONCLUSION Dietary STE supplementation promoted feed intake through the regulation of the hypothalamic neuroactive ligand‐receptor interaction pathway and the alteration of intestinal microbiota composition. This study provides valuable information about the sweetness‐independent mechanism of the STE‐induced orexigenic effect using the broiler chicken (which is insensitive to sweetness) as the animal model. © 2020 Society of Chemical Industry