Objectives: The aim of this study was to observe the depression-like behavior changes of rats exposed to lead with or without probiotic intervention, and to investigate changes in the gut microbiota and fecal short-chain fatty acids (SCFAs) levels after lead exposure, and the possible functions of probiotics in this process. Material and Methods: Adult male Sprague Dawley rats were exposed to a 300 mg/l lead acetate solution for 24 weeks, with or without probiotic (freeze-dried powder containing Lactobacillus and Bifidobacterium: 6 billion live bacteria/2 g) intervention in weeks 17–24. The sucrose preference test (SPT), the forced swim test (FST), and the tail suspension test (TST) were preformed to study the depression- like behaviors of these rats. The alteration of rat gut microbiota induced by lead exposure was analyzed by 16S rRNA sequencing, and the levels of fecal SCFAs were detected using gas chromatography. Results: Neurobehavioral tests showed that lead exposure induced depression-like behavior in rats, including reduced sucrose preference in the SPT, and increased immobility times in the FST and the TST. Sequencing and gas chromatography showed that lead exposure changed the structure and the phylogenetic diversity of the gut microbiota, as well as significantly altered the levels of SCFAs. Moreover, the depression-like behaviors, and the changes in both gut microbiota and SCFAs, could be mitigated by probiotic intervention. Conclusions: Lead exposure not only changes the structure and diversity of the gut microbiome but also affects metabolic function. Probiotic intervention may be a novel initiative for the prevention and treatment of neurological damage following lead exposure. Int J Occup Med Environ Health. 2022;35(1):95–106