Study on the Therapeutic Effect of Yu-Mu-Tiao-Shen Acupuncture on Rats with Autism Spectrum Disorder.

Acupuncture has been shown to improve symptoms of Autism Spectrum Disorder (ASD), but its underlying mechanisms remain unclear. This study aims to explore the behavioral effects of Yu-Mu-Tiao-Shen Acupuncture on ASD model rats and investigate the potential molecular mechanisms based on SFRP5, β-catenin, and GSK-3β. On gestation day 12.5, SD (Sprague-Dawley) rats were intraperitoneally injected with valproic acid (VPA), and their offspring were considered a reliable ASD rat model. The offspring were randomly assigned to the VPA group or the VPA_acupuncture group (n=7), with a normal group serving as the control. The VPA_acupuncture group underwent Yu-Mu-Tiao-Shen Acupuncture from postnatal days 8 to 28. All rats underwent behavioral testing, including social interaction, open field, and Morris water maze tests. Subsequently, hippocampal and prefrontal cortical tissues were extracted for histological analysis and RNA sequencing to assess gene expression differences across groups, as well as protein expression in hippocampal tissues. Behavioral tests demonstrated that the Yu-Mu-Tiao-Shen acupuncture technique improved spontaneous activity, abnormal social interaction, and alleviated learning and memory impairments in autistic rat models. In the VPA_acupuncture group, total travel distance, average speed, social interaction time, time spent exploring novel rats, and the number of platform crossings were significantly increased (P < 0.05). Transmission electron microscopy revealed abnormal synaptic and mitochondrial structures in the hippocampal tissue of the VPA group. In contrast, the SD and VPA_acupuncture groups showed relatively intact pre- and post-synaptic membrane structures and normal mitochondrial morphology. Differentially expressed gene (DEG) analysis revealed 111 significantly altered genes in the hippocampus and 282 in the prefrontal cortex between the VPA_acupuncture and VPA groups. Furthermore, in the VPA_acupuncture group, the hippocampal expression levels of SFRP5, β-catenin, and GSK-3β proteins were significantly reduced compared to the model group (P < 0.05). Yu-Mu-Tiao-Shen Acupuncture can regulate the expression of relevant proteins, modulating synaptic plasticity in hippocampal and prefrontal neurons, which effectively improves motor, social cognitive, and spatial memory abilities in ASD rats. This study provides valuable insights for further exploration of ASD treatments.