Multi-omics reveal gut microbiota–metabolite–adipose axis as the driver of hesperidin-mediated metabolic improvement in obese mice

Hesperidin, a citrus flavonoid glycoside, was investigated for its protective effects against high-fat diet (HFD)-induced metabolic syndrome in mice. Twelve weeks of supplementation markedly attenuated body weight gain, hepatic steatosis, adipocyte hypertrophy, dyslipidemia, and systemic inflammation, while enhancing glucose tolerance and insulin sensitivity. 16S rRNA sequencing demonstrated that hesperidin partially restored microbial diversity and selectively enriched beneficial taxa, including Lactobacillus, Bifidobacterium, and Akkermansia. Serum metabolomics revealed increased levels of microbial-derived metabolites such as cinnamic acid, hippuric acid, and sulfated phenolic acids, compounds associated with anti-obesity, antioxidant and anti-inflammatory activities. Transcriptomic profiling of inguinal white adipose tissue identified broad remodeling of metabolic pathways, with notable activation of calcium signaling, implicating both UCP1-dependent browning and UCP1-independent calcium futile cycling in thermogenesis. Importantly, antibiotic treatment abolished the metabolic benefits and suppressed the generation of bioactive metabolites, underscoring the indispensable role of gut microbiota in hesperidin bioactivity. Together, these findings delineate a microbiota–metabolite–adipose tissue axis through which hesperidin confers systemic metabolic protection, highlighting its potential as a microbiota-targeted dietary strategy for managing obesity-related disorders. Overall design: Male mice were fed a control diet (CD) or high-fat diet (HFD) with or without hesperidin supplementation (HEP) for 12 weeks. Multi-omics approaches, including 16S rRNA sequencing, untargeted serum metabolomics, and adipose transcriptomics, were employed.