<p><span style="color: black;"><em>Panax notoginseng</em> is a recognized Chinese medicinal herb widely used as a medicinal food and dietary supplement. <em>P. notoginseng</em> roots are rich in bioactive saponins with diverse pharmacological properties. Herein, the present aimed to optimize the ultrasound-assisted extraction of total saponin content from <em>P. notoginseng</em> roots using response surface methodology. Three independent variables, including EtOH concentration, extraction time, and solid-liquid ratio, were selected based on single-factor experiments. The optimal conditions were 26.7% EtOH concentration, 52.1 min, and 1/22.6 (w/v) material-to-solvent ratio. Under these conditions, the TSC reached (17.327 ± 0.184) mg OA/g extract, closely agreeing with the predicted values and confirming the reliability of the model. Moreover, the optimized extract exhibited significant antioxidant and anti-inflammatory activities. These results suggested that <em>P. notoginseng</em> root extract, obtained from UAE method, could be used in various pharmaceutical applications.</span></p>