Objective To observe the effect of electroacupuncture at Neiguan and Yanglingquan points on anxiety-like behavior and energy metabolism in the hippocampus in mice with posttraumatic stress disorder (PTSD), and to investigate the mechanism of action of electroacupuncture in improving PTSD. Methods Among 68 male C57BL/6 mice, 15 were randomly selected as normal group, and the other mice were used to establish a model of PTSD using the single prolonged stress with foot shock procedure. After successful modeling, 45 mice were randomly divided into model group, electroacupuncture group, and electroacupuncture+inhibitor group, with 15 mice in each group. After successful modeling for 7 days, the mice in the electroacupuncture group were given acupuncture at Neiguan and Yanglingquan points with continuous wave at a frequency of 2 Hz for 20 minutes each time, for 7 days, and those in the electroacupuncture+inhibitor group were given intraperitoneal injection of adenosine monophosphate activated protein kinase (AMPK) inhibitor at a dose of 50 mL/kg at 2 hours before electroacupuncture. The open field test and the elevated plus maze test were used to assess anxiety-like behavior; Western blot was used to measure the protein expression levels of AMPK, phosphorylated AMPK (p-AMPK), and peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) in hippocampal tissue; ELISA was used to measure the levels of adenosine monophosphate (AMP), adenosine triphospate(ATP), adenosine diphosphate (ADP), and adenosine triphosphatase (ATPase) in hippocampal tissue, and AMP/ATP ratio was calculated; transmission electron microscopy was used to observe the changes in the ultrastructure of hippocampal mitochondria; mitochondrial membrane potential measured by JC-1 probe was used to observe hippocampal mitochondrial injury. Results Compared with the normal group, the model group had significant reductions in total distance, the number of entries into the central field, retention time in the central field, the number of entries into the open arms, and the percentage of time spent in the open arms (P<0.05), significant reductions in the protein expression levels of p-AMPK and PGC-1α in hippocampal tissue (P<0.05), significant increases in the levels of AMP and ADP and AMP/ATP ratio (P<0.05), and significant reductions in the levels of ATP and ATPase (P<0.05), with the swelling and shrinkage of neuronal nuclei, a reduction in the number of mitochondria, mild swelling of mitochondria, and a significant reduction or disruption of mitochondrial cristae, as well as a significant reduction in mitochondrial membrane potential (P<0.05). Compared with the model group, the electroacupuncture group and the electroacupuncture+inhibitor group had significant increases in total distance, the number of entries into the central field, retention time in the central field, the number of entries into the open arms, and the percentage of time spent in the open arms (P<0.05), significant increases in the protein expression levels of p-AMPK and PGC-1α in hippocampal tissue (P<0.05), significant reductions in the levels of AMP and ADP and AMP/ATP ratio (P<0.05), and significant increases in the levels of ATP and ATPase (P<0.05); the electroacupuncture group had intact and clear nuclear membranes, a slight increase in the number of mitochondria, and alleviation of mitochondrial rupture and the reduction in mitochondrial cristae, and the electroacupuncture+inhibitor group had slight reductions in the degree of mitochondrial cristae swelling and disruption; the electroacupuncture group and the electroacupuncture+inhibitor group had a significant increase in mitochondrial membrane potential (P<0.05). Compared with the electroacupuncture group, the electroacupuncture+inhibitor group had significant reductions in total distance, the number of entries into the central field, and retention time in the central field (P<0.05), significant reductions in the number of entries into the open arms and the percentage of time spent in the open arms (P<0.05), significant reductions in the protein expression levels of p-AMPK and PGC-1α in hippocampal tissue (P<0.05), significant increases in the levels of AMP and ADP and AMP/ATP ratio (P<0.05), and significant reductions in the levels of ATP and ATPase (P<0.05), as well as a significant reduction in mitochondrial membrane potential (P<0.05). Conclusion Electroacupuncture at Neiguan and Yanglingquan points can improve anxiety-like behavior in mice with PTSD, possibly by inducing AMPK phosphorylation, activating the downstream factor PGC-1α, and improving hippocampal mitochondrial function via the AMPK/PGC-1α signaling pathway, thereby increasing the content of ATP and regulating energy metabolism.