Trophocytes and fat cells of honeybees (Apis mellifera) have served as targets for cellular senescence studies; their changes of aging-related molecules and mitochondrial energy utilization with advancing age have been clarified (Hsieh and Hsu, 2011; Chuang and Hsu, 2012). However, the changes of energy-regulated molecues with advancing age in workers are unknown. In this study, young (2-day-old) and old (50-day-old) workers were selected from the same colony in hive on the same days for the experiments. The expression of adenosine monophosphate-activated protein kinase (AMPK), phosphorylated AMPK (pAMPK), silent information regulator 1 (SirT1), and peroxisome proliferator-activated receptor-α (PPAR-α) were evaluated in the trophocytes and fat cells of young and old workers reared in a field hive by Western blotting. The activity of AMPK and SirT1 were evaluated by analytic Kits. The concentration of AMP, ADP, ATP, and AMP/ATP ratio and ADP/ATP ratio were evaluated by ultra performance liquid chromatography. The results showed that (1) AMPK expression increased with advancing age (n = 18, P < 0.01, two-sample t test), whereas pAMPK expression decreased with advancing age (n = 18, P < 0.05, two-sample t test), pAMPK/AMPK ratio decreased with advancing age (n = 18, P < 0.05, two-sample t test), and AMPK activity decreased with advancing age with 0.62 ± 0.11 and 0.14 ± 0.03 450 nm mg-1 protein in young and old workers (n = 48, P < 0.01, two-sample t test); (2) ATP concentration decreased with advancing age with 44.39 ± 3.50 and 30.51 ± 2.43 nmol mg−1 of protein in young and old workers (n = 10, P < 0.01, two-sample t test) and ADP concentration decreased with advancing age with 4.60 ± 0.39 and 2.78 ± 0.26 nmol mg−1 of protein in young and old workers (n = 10, P < 0.005, two-sample t test), whereas AMP concentration was not different with 1.45 ± 0.16 and 1.61 ± 0.28 nmol mg−1 of protein in young and old workers (n = 10, P > 0.01, two-sample t test). ADP/ATP ratio was not different with advancing age (n = 10, P > 0.05, two-sample t test), whereas AMP/ATP ratio increased with advancing age (n = 10, P < 0.005, two-sample t test); (4) SirT1 expression increased with advancing age (n = 21, P < 0.05, two-sample t test), whereas their activity decreased with advancing age with 73.2 ± 0.9 and 59.7 ± 3.7 fluorescence μg−1 of protein in the trophocytes and fat cells of young and old workers (n = 30, P < 0.01, two-sample t test); and (5) PPAR-α expression decreased with advancing age (n = 18, P < 0.01, two-sample t test). These results show that young workers express higher AMPK and SirT1 activity than old workers and that aging results in a decline in AMPK and SirT1 activity in worker honeybees.