This study investigated the mechanism of Smilacis Glabrae Rhizoma (SGR) against hyperuricemia due to dampness-heat through integrated network pharmacology, molecular docking and in vivo experiments. Potential active components, therapeutic targets, and signaling pathways of SGR against HUA were identified through network pharmacology. The binding affinities between the key compounds and core targets were validated using molecular docking. Subsequent an HUA rat model with dampness-heat syndrome was established for in vivo experimental verification. Network pharmacology analysis revealed 12 potential active components of SGR for HUA treatment, with key targets including interleukin-6 (IL-6), tumor necrosis factor (TNF), peroxisome proliferator-activated receptor gamma (PPARγ), B-cell lymphoma-2 (BCL-2), ATP binding cassette subfamily G member 2 (ABCG2), and peroxisome proliferator activated receptor alpha (PPARα). Key pathways involved included lipid metabolism, atherosclerosis, folate metabolism, adenosine monophosphate-activated protein kinase (AMPK) signaling. Molecular docking demonstrated strong binding affinity between core compounds and critical targets. Animal experiments showed that SGR significantly reduced Serum uric acid (UA) and lipid levels, upregulated renal ABCG2 and organic anion transporter 1‌（OAT1）mRNA expression, and downregulated uric acid transporter 1（URAT1） mRNA expression in model rats. Additionally, it enhanced renal expression of phosphorylated AMPK, peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), PPARγ, and ABCG2.These findings suggest that SGR mitigates hyperuricemia by promoting uric acid excretion and reducing its reabsorption. The underlying mechanism involves increasing AMPK phosphorylation, activating PGC-1α, upregulating PPARγ expression, and subsequently enhancing the downstream target ABCG2 to accelerate uric acid elimination.