Endogenous relationship to restraint and human handling were studied with growing-fattening steers. Thirty-five crossbred (Japanese Black x Holstein) steers aged 6-10 months were randomly assigned to three pens. They had free access to an Italian ryegrass hay and a restricted amount of high-concentrate diets (total digestible nutrients (TDN) 70.5%, digestible crude protein (DCP) 10.0%) for the first 6 months of trial. Then they had free access to an oat hay and another vitamin A-restricted diet (TDN 72.0%, DCP 10.0%) until slaughter. The steers were individually driven into a restraint stall, and bodyweight was recorded. Blood samples were then collected under haltered conditions. These serial handling procedures started at 2 h after the morning feeding were conducted in months 1, 3, 5 (growing stage, GS) and in months 7, 9, 11 (fattening stage, FS) of the trial. Mean peripheral blood concentrations of epinephrine (A; GS, 117.4 +/- 76.4 pg/mL; FS, 64.1 +/- 34.2 pg/mL), norepinephrine (NA; GS, 257.7 +/- 95.0 pg/mL; FS, 125.9 +/- 44.1 pg/mL), cortisol (GS, 1.6 +/- 0.8 pg/dL; FS, 1.2 +/- 0.4 pg/dL), glucose (GS, 83.1 +/- 7.5 mg/dL; FS, 71.9 +/- 6.9 mg/dL, non-esterified fatty acid (NEFA; GS, 0.13 +/- 0.06 mEQ/L; FS, 0.10 +/- 0.06 mEQ/L) and vitamin A (GS, 90.5 +/- 24.6 IU/dL; FS, 37.2 +/- 21.3 IU/dL) were higher (all P < 0.01) in the GS than in the FS, whereas those of insulin (GS, 1.06 +/- 0.82 mu U/mL; FS, 1.36 +/- 0.61 mu U/mL) and leptin (GS, 4.5 +/- 1.8 ng/mL human equivalent (HE); FS, 6.8 +/- 2.4 ng/mL HE) were lower (both P < 0.01). The metabolite that correlated with A and NA was glucose (A: r= 0.61, P < 0.001; NA: r= 0.53, P < 0.01) in the GS, and the metabolites correlating with A, NA and cortisol were NEFA (A: r=0.31, P < 0.10; NA: r=0.32, P < 0.10; cortisol: r=0.41, P < 0.05) and triglyceride (A: r=-0.37, P < 0.05; NA: r=-0.39, P < 0.05) in the FS. Vitamin A was a mediator between A (r=-0.38, P < 0.05) and NA (r=-0.42, P < 0.05) and insulin (r=0.31, P < 0.10) in the GS, and between NA (r=-0.33, P < 0.10) and leptin (r= 0.38, P < 0.05) in the FS. In conclusion, when changing from the growing to the fattening stages, the stress of handling and restraint had caused the pathways to shift from carbohydrate metabolism to lipid metabolism. In addition, vitamin A seemed to be an important mediator in the endogenous pathways in both stages.