Display omitted •Consumption of beef fat together with capsaicin inhibited dyslipidemia in mice.•Consumption of capsaicin improved oxidative stress in beef fat-fed mice.•Capsaicin elevated enteric acetic acid level in beef fat-fed mice. This study was designed to evaluate the preventive mechanisms of capsaicin on beef fat-induced lipid metabolism disorder, oxidative stress and dysbacteriosis in mice. Mice were fed a control diet or a 30% beef-fat diet (BF) with or without 0.001% capsaicin for 12 weeks. The consumption of a BF with capsaicin effectively inhibited obesity and dyslipidemia in mice. Consumption of capsaicin improved the oxidative stress by reducing the hepatic malonaldehyde content and elevating the glutathionperoxidase activity. GC–MS analysis showed that the acetic acid content in the cecum was significantly increased by capsaicin supplementation. The enteric microflora species was not changed significantly by the consumption of capsaicin in BF-fed mice. The alleviation of dyslipidemia might be because capsaicin enhanced the antioxidant activity and elevated the enteric acetic acid levels in high beef fat-fed mice. Our results suggest that the ingestion of capsaicin might be an effective strategy for preventing beef fat-induced hyperlipidemia.