中国现代中药2025，Vol.27Issue(4)：685-694,10.DOI:10.13313/j.issn.1673-4890.20240730003
基于cAMP/PKA/CREB通路的神牡安神胶囊延长深度睡眠作用机制探讨
Shenmu Anshen Capsules Prolong Deep Sleep via cAMP/PKA/CREB Pathway
摘要
Abstract
Objective:To study the effects of Shenmu Anshen Capsules in improving the deep sleep of the mouse model of p-chlorophenylalanine(PCPA)-induced insomnia.Methods:The mouse model of insomnia was established by intraperitoneal injection of PCPA in C57 male mice,and mice were randomized into control,model,diazepam,and low-,medium-,and high-dose(0.41,1.23,and 3.69 g·kg-1,respectively)Shenmu Anshen Capsules groups.Mice were administrated with drugs for 7 consecutive days after modeling.The mental state and body weight of mice in each group were observed,and the open field test and elevated plus maze test were conducted to evaluate the spontaneous activities reflecting the anxiety status of mice in each group.Morphologic changes of neurons in the hippocampus were observed by hematoxylin-eosin staining.The levels of gamma-aminobutyric acid(GABA),dopamine(DA),cortisol(Cort),melatonin(MT),cyclic adenosine monophosphate(cAMP),and brain-derived neurotrophic factor(BDNF)in the brain tissue of mice were measured by enzyme-linked immunosorbent assay.The protein levels of GABA A receptor subunit β3(GABRA3),protein kinase A(PKA),and cAMP-response element binding protein(CREB)in the brain tissue of mice were determined by Western blot.Electronencephalography(EEG)and electromyography(EMG)were employed to analyze the sleep structure of mice within 8 h after administration with Shenmu Anshen Capsules.The awake duration,non-rapid eye movement(NREM)sleep duration and rapid eye movement(REM)sleep duration of mice in each group were recorded.Results:The successfully modeled mice showed irritability and circadian rhythm disruption.Compared with the control group,the model group showed reductions in the number of crossings(P<0.05),time in the central zone(P<0.05),total movement distance(P<0.05),and movement distance in the central zone(P<0.01),and prolonged total residence time(P<0.01)in the open field test.In the elevated plus maze test,the model group showed decreases in the percentage of mice entering the open arm and the percentage of time in the open arm(P<0.01),lowered levels of GABA,MT,BDNF,and cAMP(P<0.05,P<0.01)and elevated levels of Cort and DA(P<0.05)in the brain tissue,and down-regulated protein levels of GABRA3,PKA,and CREB in the brain tissue(P<0.05).In addition,the model group had prolonged awake duration(P<0.05)and reduced NREM and REM sleep duration(P<0.05).Compared with the model group,Shenmu Anshen Capsules and diazepam groups showed increases in the number of crossings(P<0.05,P<0.01),movement distance in the central zone and total movement distance(P<0.01),and residence time in the central zone(P<0.05),and a reduction in the total residence time(P<0.05,P<0.01)in the open field test.In the elevated plus maze test,the percentage of mice entering the open arm and the percentage of time in the open arm were increased in the diazepam group and Shenmu Anshen Capsules groups(P<0.05,P<0.01).Both the medium and high doses of Shenmu Anshen Capsules elevated the levels of GABA,MT,BDNF,and cAMP(P<0.01),lowered the levels of Cort and DA(P<0.01),and up-regulated the protein levels of GABRA3,PKA,and CREB in the brain tissue(P<0.05,P<0.01)compared with the model group.High-dose Shenmu Anshen Capsules reduced the awake duration(P<0.01),prolonged the NREM and REM sleep duration(P<0.01),and made the sleep structure more complete.Conclusion:Shenmu Anshen Capsules can exert sedative and hypnotic effects on PCPA-induced insomnia and alleviate insomnia-induced tension and anxiety in mice.Through regulating the secretion of endogenous substances Cort,MT,GABA,DA,and BDNF and affecting the cAMP/PKA/CREB pathway,Shenmu Anshen Capsules prolong the total sleep duration,NREM sleep duration,and REM sleep duration,thereby improving the sleep structure,exerting sedative and hypnotic effects,and prolonging deep sleep.关键词
神牡安神胶囊/失眠/神经衰弱/镇静催眠/深度睡眠/环磷酸腺苷/蛋白激酶A/环磷酸腺苷反应元件结合蛋白信号通路Key words
Shenmu Anshen Capsules/insomnia/neurasthenia/sedation and hypnosis/deep sleep/cyclic adenosine phosphate(cAMP)/protein kinase A(PKA)/cAMP-response element binding protein(CREB)signaling pathway分类
中医学引用本文复制引用
尤诗晴,张薇,徐畅,陈静婷,肖驿泽,刘莹,杨晓菲,吴嘉怡,王金朝,褚福浩..基于cAMP/PKA/CREB通路的神牡安神胶囊延长深度睡眠作用机制探讨[J].中国现代中药,2025,27(4):685-694,10.基金项目
北京中医药大学青年科学家培育计划项目(BUCM-2019-QNKXJC008) （BUCM-2019-QNKXJC008）
国家中医药管理局高水平中医药重点学科建设项目(zyyzdxk-2023258) （zyyzdxk-2023258）
