茯神水煎液对PCPA致失眠小鼠5-HT系统和GABA表达的影响  

Effect of Fushen Decoction on 5-HT system and GABA expression in mouse model of PCPA-induced insomnia

作　　者：胡俊航 许菲 王桐生[1] 彭华胜[2] 李莉[1] HU Jun-hang;XU Fei;WANG Tong-sheng;PENG Hua-sheng;LI Li(Department of Pharmacology,School of Integrated Chinese and Western Medicine,Anhui University of Chinese Medicine,Hefei 230012,China;National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]安徽中医药大学中西医结合学院药理学教研室,安徽合肥230012 [2]中国中医科学院中药资源中心,北京100700

出　　处：《中国中药杂志》2025年第6期1581-1591,共11页China Journal of Chinese Materia Medica

基　　金：国家重点研发计划项目(2022YFC3500903)。

摘　　要：观察茯神水煎液对小鼠的镇静安神作用,并探讨其对对氯苯丙氨酸(PCPA)致失眠小鼠脑内5-羟色胺(5-HT)系统及γ-氨基丁酸(GABA)的影响。采用ICR小鼠,观察10、20、40 g·kg^(-1)茯神水煎液对正常小鼠和咖啡(1 g·kg^(-1),连续3 d)致兴奋小鼠自主活动的影响,以及对PCPA(350 mg·kg^(-1),连续3 d)致失眠小鼠自主活动和阈上剂量戊巴比妥钠协同睡眠的影响。对PCPA致失眠小鼠,采用酶联免疫吸附测定(ELISA)法检测血清中色氨酸羟化酶(TPH)、5-羟色氨酸(5-HTP)、5-HT水平以及脑干、海马和皮层中5-HTP、5-HT水平,免疫荧光法检测中缝核、海马和皮层中5-HT荧光强度,Western blot法检测脑干、海马和皮层中色氨酸羟化酶-2(TPH2)、5-羟色胺1A受体(5-HT_(1A)R)蛋白表达;另外,分别采用ELISA和免疫组化法检测下丘脑、海马、皮层中GABA水平。结果显示,20、40 g·kg^(-1)茯神水煎液可减少正常小鼠、咖啡致兴奋小鼠和PCPA致失眠小鼠自主活动,延长PCPA致失眠小鼠阈上剂量戊巴比妥钠睡眠持续时间。20、40 g·kg^(-1)茯神水煎液可提高PCPA致失眠小鼠血清中TPH、5-HTP和5-HT水平,上调脑干、海马和皮层中TPH2、5-HTP、5-HT和5-HT_(1A)R水平,上调下丘脑、皮层和海马中GABA表达。20、40 g·kg^(-1)茯神水煎液对小鼠有镇静安神作用,作用机制可能与上调脑内TPH2表达,增强5-HT系统,同时提高脑内GABA水平有关。This study aims to observe the mind-tranquilizing effect of Fushen Decoction on mice and investigate its effects on the 5-hydroxytryptamine(5-HT)system andγ-aminobutyric acid(GABA)in the brain of the mouse model of 4-chloro-DL-phenylalanine(PCPA)-induced insomnia.ICR mice were administrated with coffee(1 g·kg^(-1))for 3 days,and the effects of Fushen Decoction(10,20,and 40 g·kg^(-1))on the autonomic activities of normal mice and coffee-treated mice were observed.Furthermore,the effects of Fushen Decoction on the autonomic activity and sleep induced by a suprathreshold dose of pentobarbital sodium in the mouse model of PCPA(350 mg·kg^(-1)for 3 consecutive days)-induced insomnia were observed.The levels of tryptophan hydroxylase(TPH),5-hydroxytryptophan(5-HTP),and 5-HT in the serum,as well as those of 5-HTP and 5-HT in the brain stem,hippocampus,and cortex,were measured by enzyme-linked immunosorbent assay(ELISA).The fluorescence intensity of 5-HT in the raphe nucleus,hippocampus,and cortex was measured by the immunofluorescence method.The protein levels of tryptophan hydroxylase-2(TPH2)and 5-HT_(1A)receptor(5-HT_(1A)R)in the brain stem,hippocampus,and cortex were measured by Western blot.The levels of GABA in the hypothalamus,hippocampus,and cortex were measured by ELISA and immunohistochemistry methods.The results showed that Fushen Decoction(20,40 g·kg^(-1))reduced the number of autonomous activities in normal mice,coffee-treated mice,and the mouse model of PCPA-induced insomnia,and prolonged the duration of sleep induced by a suprathreshold dose of pentobarbital sodium in the mouse model.Fushen Decoction(20,40 g·kg^(-1))elevated the levels of TPH,5-HTP,and 5-HT in the serum,and TPH2,5-HTP,5-HT,and 5-HT_(1A)R in the brain stem,hippocampus,and cortex,and up-regulated GABA expression in the hypothalamus,cortex,and hippocampus of the mouse model of PCPA-induced insomnia.In conclusion,Fushen Decoction(20,40 g·kg^(-1))exerted a mind-tranquilizing effect on mice by up-regulating the expression of TPH2,enhancing

关 键 词：茯神水煎液 镇静安神 对氯苯丙氨酸(PCPA) 5-羟色胺(5-HT) 色氨酸羟化酶-2(TPH2) γ-氨基丁酸(GABA) 

分 类 号：R285.5[医药卫生—中药学]