This study aimed to assess whether the blood glucose levels influence cardiac autonomic modulation under fasting and after carbohydrate overload conditions. Participants (n = 108) were separated into lower blood glucose and higher blood glucose groups, based on the median (90.5 mg.dL(-1)) of fasting glucose assessed. The SD2, SDNN, LF indices, and LF/HF increased, and HF decreased after dextrose overload compared with fasting (p < 0.05). Body mass (78.9 vs 69.7 kg), abdominal circumference (90.2 vs 82.2 cm), systolic (113 vs 108 mm Hg) and diastolic (72 vs 67 mm Hg) blood pressure were higher (p < 0.05) in the higher blood glucose group. Heart rate variability (HRV) indices (SD1: 21.0 vs 26.5; SD2: 76.8: vs 86.1; RMSSD: 28.7 vs 37.5; SDNN: 56.1 vs 62.5 ms; pNN50: 10.6 vs 18.9%, HF: 328.4 vs 506.0; LF: 982.8 vs 1259.0 ms(2)), and the area under the curve of these indices after dextrose overload were lower in the higher blood glucose group (p < 0.05). Additionally, glycemia after dextrose overload was correlated with HRV indices (rho = -0.216 to -0.273, p < 0.05). Individuals with higher blood glucose, even in the normality range, showed impairment in the cardiac autonomic modulation both at fasting and after carbohydrate overload. Novelty: Higher fasting blood glucose impairs cardiac autonomic modulation. Carbohydrate overload impairs cardiac autonomic modulation.