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Saccharomyces cerevisiae cell wall extract (SCCWE) contains a variety of bioactive compounds, yet its antidiabetic action mechanisms remain unclear. The current work aimed to characterize the chemical components using GC-MS of SCCWE along with its antidiabetic, hepatoprotective, antioxidant, and microbiome-modulating properties in streptozotocin (STZ)-induced diabetic rats. The rats were treated with glibenclamide, SCCWE (25, 50, or 100 mg/kg), or non-diabetic normal rats as a control. Key regulators (P-AMPK, HMGR, SREBP-1c, and LXRα) as well as metabolic parameters, oxidative and inflammatory indicators, and histopathology and FTIR analysis were evaluated. Trehalose (16.03%), turanose (15.05%), glycerol (12.24%), and mannobiose (7.38%) were found to be the primary constituents by GC-MS profiling. STZ elevated fasting glucose 1.5-fold and reduced lactic acid bacteria 6.6-fold. SCCWE lowered glucose by 27.4-30.4% and restored lactic acid bacteria by 266.7-711.6%. Serum ALT, increased 2.1-fold in diabetic rats, decreased by 35.3-55.6% with SCCWE. Dyslipidemia improved markedly, with total lipids, cholesterol, and triglycerides reduced by up to 45.6%, 63.9%, and 46%. SCCWE decreased hepatic MDA by 56.5% and increased GSH up to 607.2%. It elevated P-AMPK while suppressing HMGR (18.9-154.6%), SREBP-1c (29.7-92.6%), and LXRα mRNA (21.3-87.6%). Histopathology and FTIR confirmed tissue and membrane restoration. SCCWE demonstrates potent antidiabetic and hepatoprotective activities, supporting its potential as a natural therapeutic for diabetes.