This study was conducted to investigate the relationship between total pericardial adipose tissue, liver fat and insulin resistance during diet-induced weight loss. A total of three overweight and twenty-seven obese premenopausal White women (body mass index: 28.2-46.8 kg/m2) aged 22-41 years were recruited. During 14.2±4 weeks of weight loss intervention, subjects received a low-calorie, nutritionally-balanced diet containing 800-1000 kcal/day. Results revealed that at baseline, the closest association was found between pericardial adipose tissue and intra-abdominal (visceral) adipose tissue. However, pericardial adipose tissue was also correlated with liver fat, HOMA-IR and M value. During weight loss, body weight decreased by -8.5%. Comparisons of the relative decrease in different adipose compartments showed that the highest relative loss was observed for liver fat and the lowest was for pericardial adipose tissue. With weight loss, the decrease in pericardial adipose tissue mainly depended on initial pericardial adipose tissue and on the loss in intra-abdominal adipose tissue, but did not correlate with decreases in liver fat nor with parameters of insulin sensitivity. In contrast, changes in liver fat were correlated with changes in intra-abdominal adipose tissue and in insulin sensitivity. Thus, these results suggest that decreases in pericardial adipose tissue are not associated with insulin resistance or liver fat, only with baseline pericardial adipose tissue and the loss of intra-abdominal adipose tissue.