北京中医药大学学报2025，Vol.48Issue(1)：138-148,11.DOI:10.3969/j.issn.1006-2157.2025.01.016
耳穴疗法对失眠模型大鼠睡眠的改善作用及对γ-氨基丁酸能系统通路的影响
Effect of auricular therapy on sleep improvement and the GABAergic system pathway in an insomnia rat model
摘要
Abstract
Objective To investigate the effect of auricular therapy on sleep improvement and the GABAergic system pathway in a rat model of insomnia and to explore its possible mechanism.Methods According to the random number table,60 male SD rats were randomly divided into blank control,model,auricular point sticking,auricular bloodletting,and auricular bloodletting combined with sticking groups,with 12 rats per group.Insomnia was induced by intraperitoneal injection of p-chlorophenylalanine.After establishing the insomnia model,36 rats were treated once a day with auricular point sticking or bloodletting for 5 consecutive days.After the intervention,the general condition and body weight of rats were observed;the righting reflex test was used to detect the sleep latency and duration;HE staining was used to observe the morphology of hypothalamic neuron cells;and an enzyme-linked immunosorbent assay was used to detect the GABA and glutamate content in rat serum.Immunohistochemistry(IHC)and real-time fluorescence quantitative PCR were used to detect GABA ARα1 and GABA ARγ2 protein and mRNA expression in the hypothalamus of rats,and Western blotting(WB)was used to detect GABA ARα1,GABA ARγ2,GAD65/67,GAT-1,and GABA-T protein expression in the hypothalamus of rats.Results Compared with the blank control group,the model group had a lower body weight,a significantly shorter sleep duration(P<0.05),severe damage to the morphological structure of hypothalamic neurons with disordered cell arrangement,larger intercellular gaps,enlarged cell bodies,and a vacuolated appearance.All the intervention groups had significantly higher body weight and longer sleep duration than the model group(P<0.05).Compared with the other intervention groups,the auricular point sticking group had a longer sleep duration(P<0.05),and the hypothalamic neuron cells in all intervention groups improved,with the auricular point sticking group showing more apparent improvement.The model group had a lower GABA and higher glutamate contents,and GABA ARα1,GABA ARγ2,and GAD65/67 protein expression in the hypothalamus were lower than in the blank control group.In contrast,GAT-1 and GABA-T protein expression was higher,and GABA ARα1 and GABA ARγ2 mRNA expression was lower(P<0.05).The serum GABA content in the auricular point sticking and auricular bloodletting groups was higher,and the serum glutamate content in the auricular point sticking and auricular bloodletting combined sticking groups was lower than in the model group.GABA ARα1 mRNA expression in the hypothalamus of each intervention group was significantly increased,and GABA ARγ2 mRNA expression in the hypothalamus of the auricular point sticking and auricular bloodletting combined sticking groups increased.GABA ARα1(IHC,WB),GABA ARγ2(WB),and GAD65/67 protein expression in the hypothalamus of the auricular point sticking group increased,whereas GAT-1 and GABA-T protein expression decreased.GABA ARα1 and GABA ARγ2 protein expression(IHC,WB)in the hypothalamus of the auricular bloodletting group increased,whereas GABA-T protein expression decreased.GABA ARγ1(IHC)and GABA ARγ2(WB)protein expression in the hypothalamus of the auricular bloodletting combined sticking group increased,whereas GAT-1 and GABA-T protein expression decreased(P<0.05).Compared with in the inventation groups,the serum GABA content in the auricular point sticking group increased,the serum glutamate content decreased,GABA ARα1 and GABA ARγ2 mRNA expression in the hypothalamus increased,and GABA ARα1(IHC),GAD65/67 protein expression increased.In contrast,GABA-T protein expression decreased(P<0.05),and GABA ARγ2 protein expression(IHC)in the hypothalamus of the auricular bloodletting group increased(P<0.05).Conclusion Auricular therapy,particularly auricular point sticking,may have modulated the GABAergic system pathway by upregulating hypothalamic GABA ARα1,GABA ARγ2,and GAD65/67 protein expression while downregulating GAT-1 and GABA-T protein expression to alleviate symptoms in an insomnia rat model.关键词
失眠/耳穴疗法/耳穴贴压/耳穴刺络/γ-氨基丁酸/神经递质/大鼠Key words
insomnia/auricular therapy/auricular point sticking/auricular bloodletting/γ-aminobutyric acid/neurotransmitter/rats分类
医药卫生引用本文复制引用
陈豪,佘延芬,张细芬,王雪松,高渊博,李旭欣,郑习慧,王羽,郑晓军,李海平..耳穴疗法对失眠模型大鼠睡眠的改善作用及对γ-氨基丁酸能系统通路的影响[J].北京中医药大学学报,2025,48(1):138-148,11.基金项目
国家重点研发计划项目(No.2022YFC3500601) （No.2022YFC3500601）
河北中医药大学研究生创新资助立项项目(No.XCXZZBS2024003) National Key R&D Program of China(No.2022YFC3500601) （No.XCXZZBS2024003）
