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Cage layer fatigue is a common metabolic disease associated with a calcium and phosphorus imbalance, but recently we found this disease can be led by high fat diet. In order to elucidate the pathogenesis induced by a high fat diet, we randomly divided 88 White Shell Roman layers into 2 groups. There were 44 layers in each group. The control group was fed by a standard layer rations, and the high fat group was fed by completed rations mixing with 3% soybean oil. This study successfully constructed an animal model of osteoporosis caused by high fat. Bone samples were collected for bone mineral density, bone biomechanical properties which are all decreased at 26, 30, 34, and 38 wk old. We found the pathway of tryptophan-ERK-CREB from the perspective of metabonomics which promote the bone absorption. By metabolomics, we screened the significantly activated tryptophan pathway in high fat feed and detected the elevated tryptophan metabolite serum 5-HT at 26, 30, 34 and 38 wk old in the high fat group. At 38 wk old, we detected significantly elevated protein and mRNA levels of ERK/CREB/C-fos in bone tissue in the high fat group. So we concluded that high-fat were associated with a decrease in bone density and bone biomechanical index by disrupting tryptophan-5-HT-ERK1/2-CREB metabolism signaling pathways., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)