Purpose <p>Appropriate physical exercise could prevent, delay, or reverse cognitive decline in aging people, which could improve their quality of life. However, the underlying mechanisms are still elusive. The present study aimed to explore the effects of 8-week Flexi-bar vibration exercise (FE) on the cognitive function in middle-aged and elderly women and uncover the underlying mechanisms.</p> Methods <p>Thirty middle-aged and elderly women were randomly assigned to the FE group or the control group (<i>n = </i>15 in each group). Executive function and real-time oxyhemoglobin concentration (oxy-Hb) changes in different cerebral regions were evaluated through the Color-Word Stroop Test and functional near-infrared spectroscopy, respectively. Circulating levels of Brain-derived neurotrophic factor (BDNF) and tumor necrosis factor-alpha (TNF-α) were measured by enzyme-linked immunosorbent assay. The expression levels of microRNAs were determined using microRNA sequencing and real-time PCR approaches, followed by an integrated bioinformatics/biostatistics analysis.</p> Results <p>Our data showed that 8 weeks of FE had a positive effect on preventing cognitive decline, with improved blood oxygen content in the dorsolateral prefrontal cortex (DLPFC), up-regulated BDNF level, and down-regulated level of TNF-α in serum. Besides, our data showed that 14 microRNAs, which are related to the cognitive function, were up/down-regulated in the FE group.</p> Conclusion <p>Our study revealed that 8-week FE could regulate the expression levels of microRNAs related to cognitive function, elevate BDNF level, and decrease TNF-α level, and thus improve cognitive performance in middle-aged and elderly women.</p>