关键词: 失眠, miR-124, PI3K/AKT/NF-κB信号通路, 炎症因子, 大鼠, 睡眠-觉醒周期

Abstract: Objective To explore the mechanism of miR-124 in regulating the PI3K/AKT/NF-κB signaling pathway in insomnia,to open up new ideas for the construction of an insomnia prevention and treatment system. Methods Thirty-two Sprague-Dawley rats were randomly divided into the Con group,SD group,SD miR-124 agomir group,and SD miR-124 antagomir group.HE staining was used to observe the pathological changes of the hippocampus of rats in each group.The ultrastructure of neurons and synapses was observed by transmission electron microscopy.The expression levels of miR-124 in serum and miR-124,PI3K,and AKT in hippocampal tissues were detected by qRT-PCR.ELISA was used to detect the contents of serum interleukins IL-6,IL-1β,and TNF-α in rats,and Western blotting(WB)was used to detect the expression levels of PI3K,AKT,p-AKT,and NF-κB proteins. Results (1)Body weight changes:Compared with the Con group,after modeling,each treatment group showed varying degrees of body weight loss(all P<0.05),with the SD miR-124 antagomir group showing the most significant weight reduction;(2)Sleep phases:EEG/EMG results showed that with increased sleep deprivation time,the duration of WAKE decreased,while the durations of NREM and REM increased relatively(all P<0.05).Compared with the SD group,the SD miR-124 agomir group had a larger increase in REM percentage,and a smaller NREM increase;(3)HE staining:Neuronal death and inflammatory cell infiltration of varying degrees were observed in the CA1,CA3,and DG regions of each treatment group,with the SD miR-124 antagomir group being the most severe,and the SD miR-124 agomir group being milder.Transmission electron microscopy showed mitochondrial swelling,increased autophagosomes,nuclear membrane wrinkling,or endoplasmic reticulum expansion in each treatment group;(4)ELISA results:Compared with the Con group,serum levels of IL-6,TNF-α,and IL-1β increased in the SD group(all P<0.05);compared with the SD group,the SD miR-124 antagomir group had increased inflammatory factors,while the SD miR-124 agomir group showed decreased levels;(5)qRT-PCR results:Compared with the Con group,levels of miR-124,PI3K,and AKT in hippocampal tissue decreased in the SD group;compared with the SD group,PI3K and AKT levels decreased in the SD miR-124 antagomir group,whereas PI3K and AKT levels increased in the SD miR-124 agomir group;(6)WB results:Compared with the Con group,expressions of PI3K and p-AKT proteins in the hippocampus of the SD group significantly decreased(all P<0.05).Compared with the SD group,PI3K and p-AKT levels in the SD miR-124 antagomir group decreased further(all P<0.05),whereas levels of PI3K,AKT,and p-AKT increased in the SD miR-124 agomir group(all P<0.05).Compared with the Con group,NF-κB protein expression increased in the hippocampus of the SD group,SD miR-124 antagomir group,and SD miR-124 agomir group,with the SD miR-124 antagomir group showing the greatest increase. Conclusion This study first predicted the potential association between miR-124 and insomnia,as well as its pathway,through bioinformatics analysis,leading to subsequent experiments to verify that miR-124 can improve the accumulation of inflammatory factors and changes in sleep phases in rats caused by sleep deprivation.This may be related to the regulation of the PI3K/AKT/NF-κB signaling pathway,exerting anti-inflammatory and antioxidant effects.

Key words: insomnia, miR-124, PI3K/AKT/NF-κB signaling pathway, inflammatory cytokines, rats, sleep-wake cycle