“神宁藏香”对失眠大鼠的行为学及GABA能系统通路的影响  

Effects of “Tibetan Incense” on Behavior and Gamma-aminobutyric Acid System of Insomniac Rats

作　　者：谭丽博 于佳慧 郑骁洋 刘燕 于雪 戴宁 张蔚 仁青加[2] 李峰 Tan Libo;Yu Jiahui;Zheng Xiaoyang;Liu Yan;Yu Xue;Dai Ning;Zhang Wei;Ren Qingjia;Li Feng(School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China;Tibet Institute of Tibetan Medicine,Lhasa 850000,China)

机构地区：[1]北京中医药大学中医学院,北京100029 [2]西藏藏医学院藏医药研究所,拉萨850000

出　　处：《世界科学技术-中医药现代化》2020年第11期4060-4068,共9页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：西藏自治区科技厅科技计划项目(2015XZ0IG63):基于GABA递质探讨“神宁藏香”治疗失眠的神经生物学机制,负责人:仁青加;西藏藏医药大学藏医药区域协同创新中心项目(2018XTCX011):藏医复方宁神精油治疗失眠的疗效及生物学机制研究,负责人:李峰。

摘　　要：目的观察"神宁藏香"对失眠大鼠行为学及GABA能系统通路的相关受体及递质的影响。方法采用SD大鼠腹腔注射对氯苯丙氨酸(para-chlorophenylalanine,PCPA)复制失眠模型,随机分组为模型组、阳性药组、"神宁藏香"组和安神中药组,并建立空白对照组,每组进行相应药物干预7天,模型组和空白组7天提供生理盐水,观察并记录各组大鼠的一般活动状态。通过行为学实验(翻正实验、旷场实验、悬尾实验、强迫游泳实验)对比各组之间的差异;运用Western Blot分子生物学技术探讨失眠的发病机制及"神宁藏香"对失眠大鼠GABA能系统通路中合成代谢:GAD65、GABA-T,转运:GAT-1,传导:NKCCl的蛋白含量的表达影响。结果 "神宁藏香"可以明显增加睡眠持续时间(P <0.01),显著降低旷场实验中央格停留时间(P <0.01)及增加站立次数(P <0.01),有效降低悬尾实验中不动时间(P <0.01),显著降低强迫游泳实验漂浮时间(P <0.01);"神宁藏香"可以提高下丘脑中GAD65、GAT-1蛋白的表达,降低下丘脑中GABA-T、NKCC1蛋白的表达。结论 "神宁藏香"能够有效延长失眠大鼠的睡眠持续时间,改善失眠所产生的抑郁、紧张等负性情绪,增加其空间探索的兴趣;并且其作用靶点主要是通过促进失眠大鼠下丘脑中GABA合成蛋白GAD-65的表达和降低转运蛋白NKCC1的表达而发挥治疗作用,这也为临床中推广使用"神宁藏香"治疗失眠提供参考。Objective To observe the effects of"Tibetan incense"on the behavior and relevant receptors and transmitters of the Gamma-aminobutyric acid system(GABA) of insomniac rats. Methods The insomnia rat model was established by intraperitoneal injection of Para-chlorophenylalanine(PCPA) in SD rats. The rats were randomly divided into insomnia model group, positive group,"Tibetan incense"group, tranquilizing herb group, and blank control group.Each group was given corresponding drugs for 7 days. The rats of the insomnia model group and blank control group were intragastrically infused with normal saline for 7 days, then the general activity of rats in each group was observed. The behavioral experiments(correction test, open field test, tail suspension test, forced swimming test) were conducted to observe differences between groups, and the Western Blot molecular biology technique was used to explore the pathogenesis of insomnia and the effect of"Tibetan incense"on the protein content of synthetic metabolism in GABA acid system pathway: GAD65, GABA-T, transport: GAT-1, and conduction: NKCC1. Results "Tibetan incense"could significantly increase the amount of sleep(P < 0.01), reduce the time of rats staying in the center grille in the open field test(P < 0.01), increase standing times(P < 0.01), reduce the immobility time in the tail suspension test(P < 0.01), and reduce the floating time in the forced swimming test(P < 0.01)."Tibetan incense"could increase the protein expression of GAD65 and GAT-1 and decrease the protein expression of GABA-T and NKCC1 in hypothalamus. Conclusion"Tibetan incense"has more significant advantages in prolonging sleep time, and improve the depressive state of insomniac rats, which may play a therapeutic role by up-regulating GAD-65 and reducing NKCC1 in hypothalamus. The study provides references for the clinical use of"Tibetan incense"for insomnia.

关 键 词：失眠 神宁藏香 行为学 WESTERN BLOT GABA递质 

分 类 号：R29[医药卫生—民族医学]