<p>Cognitive impairment (CI) is considered to be the precursor stage of various dementias and neurodegenerative diseases, may have potential malignant effects on the physiological, biochemical and gut microbial health of the older adults. Wooden-toys training for the older adults can be regarded as an intellectual activity or cognitive stimulating activity. However, the associations between wooden-toys training and the gut microbiota function in older adults with CI are unknown. A single-arm intervention study was conducted among 38 male older adults aged 78.58 ± 6.42 years and an 8-week wooden-toys training intervention was performed on all subjects. Physiological indexes including body mass index (BMI), blood pressure, calf circumference and grip strength were measured baseline and after the intervention. The gut microbiome of the older adults was also tested baseline and after the intervention, multivariate data analysis and functional prediction were performed. After the intervention of wooden-toys training, with the improvement of cognitive function in the older adults, the enrichment and diversity of gut microbiome in the older adults with CI was significantly higher than that in the baseline stage (<i>P</i> = 0.0378). The contents of gut metabolites propionate, lactate, leucine, pyruvate, glycerol, o-glycan and the related functions were significantly increased. Positive associations between wooden-toys training and the gut microbiota functions in male older adults with CI have been found in this study. The gut-brain axis plays a key role in this process, gut probiotics and amino acids are potential key association factors for elucidating the mechanisms that need to be verified in further studies.</p>