Aluminum is the most abundant environmental pollutant. Recent studies suggest that aluminum exposure increases the risk of multiple diseases, including intestinal barrier dysfunction. We investigated whether Pueraria lobata extract (PLE) is effective in safeguarding against aluminum chloride exposure-exacerbated intestinal barrier dysfunction. Using an experimental colitis model of aluminum-exacerbated dextran sulphate sodium (DSS)-treated mice, clinical and pathological evidence suggested that the administration of PLE counteracted aluminum exposure-induced intestinal barrier damage. In addition, we found that aluminum toxicities, including loss of tight junction molecules (TJs), upregulated pro-inflammatory cytokines, and enhanced myeloperoxidase (MPO) activity, were significantly suppressed by PLE administration. Furthermore, PLE administration was identified to inhibit activation of MAPKs and NF-κB signal pathways, which contribute to upregulation of myosin light-chain kinase (MLCK) in inflamed intestine. Taken together, these results suggest that PLE might be a potential candidate for aluminum exposure-related intestinal barrier dysfunction.