无忧汤通过RhoA/ROCK1/Drp1通路对慢性睡眠剥夺模型大鼠海马神经元细胞凋亡及线粒体损伤的影响OA

This study investigated the protective effects and underlying mechanisms of Wuyou Decoction(WYD)against hippocampal neuronal apoptosis and mitochondrial damage in a rat model of chronic sleep deprivation(CSD).Fifty male Sprague-Dawley(SD)rats were randomly assigned to the following groups:normal,model,and WYD low-,medium-,and high-dose groups(WYD-L,WYD-M,WYD-H),respectively.Except the normal group,CSD was induced in other groups using the horizontal platform sleep deprivation method,which included 7 days of adaptive modeling followed by 21 days of formal modeling.Intragastric administration of WYD was initiated concurrently with the formal modeling phase and maintained for 28 days.Learning and memory abilities were assessed using the Morris water maze test.The morphological and quantitative changes of hippocampal neurons were evaluated by hematoxylin-eosin(HE)staining,Nissl staining,and neuron-specific nuclear protein(NeuN)immunohistochemistry.Mitochondrial ultrastructure was examined by transmission electron microscopy,and adenosine triphosphate(ATP)content in the hippocampus was measured with a commercial assay kit.Western blot was employed to detect apoptosis-related proteins B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),caspase-3,cleaved caspase-3,cytochrome C(Cyt C),as well as Ras homolog family member A(RhoA),Rho-associated protein kinase 1(ROCK1),dynamin-related protein 1(Drp1),and p-Drp1(Ser616)in hippocampal tissues.Molecular docking was used to evaluate the interactions between active components of WYD and pathway-related proteins.The results revealed that,compared to the normal group,the model group showed significant cognitive impairment,neuronal damage,reduced neuronal count,and abnormal mitochondrial structure,including disrupted cristae and myelin-like figures.The ATP content decreased,along with increased Cyt C release,upregulation of Bax and cleaved caspase-3/caspase-3,downregulation of Bcl-2,and activation of the RhoA/ROCK1/Drp1 signaling pathway.In contrast,the WYD-M group exhibited improved cognitive performance,restored neuronal morphology and quantity,and alleviated mitochondrial damage,while the ATP levels also increased.Furthermore,WYD-M treatment significantly suppressed Cyt C release,downregulated Bax and cleaved caspase-3/caspase-3 expression,upregulated Bcl-2,and effectively inhibited the activation of the RhoA/ROCK1/Drp1 signaling pathway.Molecular docking indicated that paeoniflorin and ferulic acid,the primary active components of WYD,had strong binding affinities with the key proteins in the RhoA/ROCK1/Drp1 signaling pathway,with binding energies all below-5 kcal·mol-1.In conclusion,WYD may protect mitochondrial integrity,reduce Cyt C release,inhibit hippocampal neuronal apoptosis,and ultimately improve cognitive function in CSD model rats potentially by suppressing the RhoA/ROCK1/Drp1 signaling pathway.