Royal Jelly Enhances the Social Status of Submissive Rats by Restoring Balance to the Disturbed Gut–Brain Communication

Royal jelly (RJ) has long been considered a crucial dietary component in dictating caste differentiation in honeybees. As a nutritional additive, royal jelly imparts a broad range of benefits to mammals and humans; however, its precise impact on the social hierarchy of these advanced animals is not yet fully understood. This study aims to determine whether the benefits of royal jelly can be transferred to rats to alter their social ranks and uncover the underlying mechanisms. A submissive model was established by inducing dysbiosis in rats, via the persistent exposure of vancomycin. Royal jelly at a dose of 2.5 g/kg was daily administered to the subject rats during postnatal weeks (PNW) 6 and 7. At the end of the intervention, animals were subjected to agonistic, water and tube competition tests, in order to assess their dominance status. As revealed by the results, the RJ treatment significantly improved the social rank of the dysbiotic rats, demonstrating that RJ can elicit positive effect on the social behaviors (caused by dysbiosis) of rats. All behavioral paradigms yielded consistent results, with no notable differences in body weight or anxiety levels. Regarding gut microbiome, vancomycin exposure caused the dysbiosis of the subject rats, which was partially reversed by treatment with royal jelly. Specifically, the intestinal presence of Proteobacteria was profoundly attenuated by the RJ supplementation, resulting in a comparable level with the intact/dominant rats. At the genus level, both <i>Escherichia</i> and <i>Clostridium</i> displayed similar dynamics in relation to Proteobacteria, implying their involvement with the RJ-mediated dominance switching. Transcriptomic analysis in the medial prefrontal context showed that the expression of a broad range of genes was influenced by RJ intake, embodying various pathways related to neuronal transmission such as neuroactive ligan-receptor interaction, the synaptic vesicle cycle, etc. By virtue of correlation analysis, <i>Escherichia</i>, <i>Akkermansia</i> and <i>Clostridium</i> were strongly associated with a set of gene modules around gastrin releasing peptide (<i>Grp</i>) and signaling pathways around <i>Rps6ka3</i>, establishing an intrinsic gut-brain communication. Furthermore, the infection trials of <i>Escherichia</i> significantly degraded the social ranks of the RJ-remedied rats in tube tests, while a series of cerebral genes like <i>Grpr</i> and <i>Grpel1</i>, as well as prefrontal spine density, were concordantly altered, underscoring the critical role of the gut-brain link in deciding the outcomes of the dyadic contests. In summary, this is an intriguing example of how royal jelly can influence the social ranks of mammals, emphasizing the importance of microbe-host interaction in mediating this species-spanning function of royal jelly in shaping social hierarchy.