OBJECTIVES This study evaluated a simple relaxation breathing exercise for acute improvement of postprandial glycemic and insulinemic status. DESIGN Healthy human subjects were randomized to control breathing (CB; n=13) or a relaxation breathing exercise (RB; n=13) that was repeated every 10 minutes for the 30 minutes before and 90 minutes after consuming a glucose challenge (oral glucose tolerance test; OGTT; 75 g/240 mL). Blood samples were collected before, and 30, 60, and 90 minutes post OGTT for glucose and insulin analysis. RESULTS Blood glucose at 0 minutes (pre-OGTT), and 30, 60, and 90 minutes post-OGTT with continued RB was 93.7±1.9, 136.5±8.1, 165.7±8.1, and 130.2±6.9 mg/dL, and 97.1±2.4, 173.1±8.4, 158.7±11.1, and 137.1±10.1 with CB, respectively. RB blood glucose was significantly lower at 30 minutes than CB. Glucose area under the curve (AUC) for CB and RB were not significantly different. Plasma insulin for both CB and RB was significantly increased relative to baseline at 30, 60, and 90 minutes. Insulin values for RB tended to be higher than CB at 30 and 60 minutes, although the difference was not statistically significant. Insulin AUC for CB and RB was not significantly different. CONCLUSIONS Relaxation breathing acutely improves the glycemic response of healthy subjects, and breathing pattern could be important for interpretation of glycemic index measurements.