Glypican-4 (GPC-4) is a novel adipomyokine that enhances insulin signaling. Glycosylphosphatidylinositol-specific phospholipase D (GPLD1) is thought to release GPC-4 and is itself an insulin-regulated enzyme. Beneficial effects of exercise training and chamomile flowers extract (CFE) are shown through activation of PPARγ, which is a promising drug target in diabetes and associated with GPC-4 synthesis. This study investigated the effects of 14-week treadmill running and CFE on serum GPC-4, GPLD1, and insulin levels in streptozotocin-nicotinamide (STZ-NA)-induced diabetic rats.Thirty-two STZ-NA-induced diabetic male Wistar rats were randomly assigned to four groups: control (C), training (T), CFE treatment (CFE), and training plus CFE treatment (TCFE) groups. The training groups were exercised on treadmill 5 days/week and the treating groups were fed with 200 mg/kg/day CFE in drinking water for 14 weeks. Finally, serum GPC-4, GPLD1, and insulin levels were analyzed via sandwich ELISA.Compared to the control group, serum insulin levels were significantly higher in the T, CFE, and TCFE groups (p < 0.05, p < 0.05, p < 0.01, respectively), while OGTT and serum GPLD1 levels were significantly lower in the T, CFE, and TCFE groups (all p < 0.001). Changes in serum GPC-4 levels were not significant. Serum GPLD1 levels were negatively correlated with insulin levels and HOMA-IS (both p < 0.001).This study suggests that endurance training and CFE may downregulate serum GPLD1 levels in STZ-NA-induced diabetic rats, which associate with the serum insulin profile. However, the results show that endurance training and CFE may not cause serum GPC-4 adaptation in STZ-NA-induced diabetic rats.