北京中医药大学学报2025，Vol.48Issue(6)：877-888,12.DOI:10.3969/j.issn.1006-2157.2025.06.020
督脉推法激活PI3K/AKT信号通路改善孤独症谱系障碍大鼠行为学的机制研究
Mechanism of governor vessel pushing manipulation activating the PI3K/AKT signaling pathway to improve behavioral outcomes in rats with autism spectrum disorder
摘要
Abstract
Objective To investigate the effects of governor vessel pushing manipulation on behavioral outcomes in valproic acid(VPA)-induced autism spectrum disorder(ASD)rats and explore its underlying mechanisms using prefrontal RNA sequencing(RNA-Seq).Methods Nine Sprague-Dawley pregnant rats at gestational day 12.5 were divided into two groups,six received intraperitoneal VPA injection(600 mg/kg)for modeling,and three received saline.Male offspring at postnatal day 21 were evaluated using the three-chamber social test and open field test to validate the ASD model.VPA-induced male offspring were randomly assigned to the model group(n=5)or tuina group(n=5),while saline offspring formed the blank group(n=5).The blank group and model group received no intervention,while the tuina group underwent governor vessel pushing manipulation stimulation along the governor vessel using a custom device,twice a day for 14 days,totaling 28 times.Post-intervention,behavioral assessments included social index(SI)and social preference index(SPI)in the three-chamber test,total distance traveled and central zone time in the open field test,marble-burying test for stereotyped behaviors,and Nissl staining for prefrontal cortical neuron survival.RNA-Seq identified differentially expressed genes(DEGs)in the prefrontal cortex,followed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses.Real time fluorogenic quantitative PCR(RT-qPCR)validated DEGs,and Western blotting analyzed proteins in enriched pathways.Results Pre-intervention,both model and tuina groups showed reduced SI,SPI,total distance,and central zone time compared to the blank group(P<0.05),confirming successful modeling.Post-intervention,the model group exhibited lower SI,SPI,total distance,central zone time,increased marble-burying(P<0.05),and fewer Nissl bodies(P<0.01)versus the blank group.Compared to the model group,the tuina group displayed improved SI,SPI,total distance,central zone time(P<0.05),reduced marble-burying(P<0.05),and increased Nissl bodies(P<0.01).RNA-Seq revealed 213 prefrontal DEGs(181 upregulated,32 downregulated)in the tuina group.GO analysis highlighted cellular components,while KEGG identified 181 pathways,with 67 significantly enriched(P<0.05),notably the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway.RT-qPCR confirmed decreased collagen type Ⅰ alpha 2(Col1α2),transforming growth factor-α(TGF-α),epidermal growth factor receptor 3(ErbB3),and serum/glucocorticoid regulated kinase 2(Sgk2)(P<0.05),and increased hepatic growth factor(Hgf)(P<0.01)in the model group,reversed by governor vessel pushing manipulation.Western blotting showed reduced prefrontal NRG1,ErbB3,nNOS,PI3K,AKT,p-nNOS,p-PI3K,and p-AKT in the model group(P<0.05),which were upregulated by tuina.Conclusion Governor vessel pushing manipulation ameliorates social deficits,anxiety,stereotyped behaviors,and neuronal loss in ASD rats,potentially via activation of the PI3K/AKT signaling pathway.关键词
推拿/督脉/孤独症谱系障碍/行为学/转录组学测序Key words
tuina/governor vessel/autism spectrum disorder/behavioral outcomes/transcriptome sequencing分类
医药卫生引用本文复制引用
冯祥,张宇星,唐丽亚,智慧,李桃,王光宇,成绍武,李江山..督脉推法激活PI3K/AKT信号通路改善孤独症谱系障碍大鼠行为学的机制研究[J].北京中医药大学学报,2025,48(6):877-888,12.基金项目
中国博士后科学基金面上项目(No.2024M760895) （No.2024M760895）
国家资助博士后研究人员计划项目(No.GZC20230784) （No.GZC20230784）
湖南省自然科学基金项目(No.2022JJ30434,No.2024JJ6341) （No.2022JJ30434,No.2024JJ6341）
湖南省教育厅科学研究项目(No.23B0390) （No.23B0390）
湖南中医药大学本科生科研创新基金项目(No.2023BKS085)China Postdoctoral Science Foundation(No.2024M760895) （No.2023BKS085）
