摩腹法结合揿针对焦虑性失眠大鼠下丘脑睡眠内稳态系统腹外侧视前核的影响  

Effect of Abdominal Massage Combined with Thumb-tack Needling for Subcutaeous Embedding on Ventrolateral Preoptic Nucleus of Hypothalamic Sleep Homeostasis System in Rats with Anxiety Insomnia

作　　者：邱思淦 张雪枫 古来撒尔·艾克拜尔 卢旭昇 郭蕾 张星平[1,2,3] 刘俊昌 QIU Si-Gan;ZHANG Xue-Feng;Gulaisaer·AIKEBAIER;LU Xu-Sheng;GUO Lei;ZHANG Xing-Ping;LIU Jun-Chang(School of Traditional Chinese Medicine,Xinjiang Medical University,Urumqi 830017 Xinjiang,China;Xinjiang Uygur Autonomous Region Academy of Traditional Chinese Medicine,Urumqi 830000 Xinjiang,China;Xinjiang Medical University Affiliated Hospital of Traditional Chinese Medicine,Urumqi 830000 Xinjiang,China)

机构地区：[1]新疆医科大学中医学院,乌鲁木齐齐830017 [2]新疆维吾尔自治区中医药研究院,乌鲁木齐830000 [3]新疆医科大学附属中医医院,乌鲁木齐830000

出　　处：《广州中医药大学学报》2024年第6期1549-1559,共11页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:81960906);新疆维吾尔自治区科技创新团队(天山创新团队)项目(编号:2022TSYCTD0008);新疆维吾尔自治区研究生科研创新项目(编号:XJ2023G199)。

摘　　要：【目的】观察摩腹法结合揿针对焦虑性失眠大鼠睡眠内稳态系统的影响。【方法】将40只大鼠随机分为正常组、模型组、摩腹组、揿针组和摩腹加揿针组,每组8只。除正常组,其余各组大鼠采用多因素复合刺激法复制焦虑性失眠模型。相应干预后,Morris水迷宫实验检测大鼠学习记忆水平,旷场实验检测焦虑应激程度,苏木素-伊红(HE)染色观察下丘脑腹外侧视前核(VLPO)神经元病理变化,免疫组织化学法、实时定量聚合酶链反应(qRT-PCR)法、Western Blot法检测VLPO脑区N-甲基-D-天冬氨酸(NMDA)受体亚基NR1、NR2B和钙调蛋白激酶Ⅱ(CaMKⅡ)的蛋白及mRNA表达。【结果】与正常组比较,模型组大鼠日间焦虑症状加重,睡眠潜伏期延长、持续时间缩短(P<0.01),水迷宫定向航行实验平均游泳总路程和平均逃避潜伏期增加(P<0.01),空间探索实验穿越隐藏平台的次数和目标象限滞留时间减少(P<0.01),旷场实验运动距离、中央格跨越格数与中央格滞留时间均显著减少(P<0.01),修饰频次与直立次数无统计学差异(P>0.05),VLPO脑区神经元出现病理损伤,NR1、CaMKⅡ蛋白及mRNA表达水平降低(P<0.01),NR2B蛋白及mRNA表达水平升高(P<0.01);与模型组比较,摩腹组、揿针组和摩腹加揿针组大鼠水迷宫实验学习记忆水平与旷场实验焦虑应激程度得到明显恢复(P<0.05或P<0.01),VLPO脑区神经元损伤改善,NR1、CaMKⅡ蛋白及mRNA表达水平升高(P<0.05或P<0.01),NR2B蛋白及mRNA表达水平降低(P<0.05或P<0.01);摩腹加揿针组上述各指标改善效果均优于摩腹组或揿针组(P<0.05或P<0.01)。【结论】摩腹法结合揿针可对焦虑性失眠大鼠起到促眠和抗焦虑的疗效,其相关机制可能与调控VLPO脑区NR1/NR2B之间的动态平衡及上调CaMKⅡ表达水平,改善VLPO脑区神经元功能,进而恢复下丘脑睡眠内稳态系统调节作用有关。Objective To observe the effect of abdominal massage combined with thumb-tack needling for subcutaeous embedding on sleep homeostasis system in rats with anxiety insomnia.Methods Forty rats were randomly divided into normal group,model group,abdominal massage group,thumb-tack needling for subcutaeous embedding group and abdominal massage plus thumb-tack needling for subcutaeous embedding group,with 8 rats in each group.Except for the normal group,the rats in the other groups were used to replicate the model of anxiety insomnia by multi-factor compound stimulation.After the corresponding intervention,Morris water maze test was used to detect the level of learning and memory.Open field test was used to detect the degree of anxiety stress.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of hypothalamic ventral lateral preoptic nucleus(VLPO)neurons.Immunohistochemistry,real-time quantitative polymerase chain reaction(qRT-PCR)and Western Blot were used to detect the protein and mRNA expression of N-methyl-D-aspartate(NMDA)receptor subunits NR1,NR2B and calmodulin kinase I(CaMK I)in hypothalamic VLPO area,respectively.Results Compared with the normal group,the daytime anxiety symptoms of the rats in the model group were aggravated,the sleep latency was prolonged and the duration was shortened(P<0.01).The average total swimming distance and average escape latency of the water maze directional navigation experiment were increased(P<0.01).The number of crossing the hidden platform and the retention time of the target quadrant in the space exploration experiment were decreased(P<0.01).The movement distance,the number of central grid crossings and the retention time of the central grid in the open field experiment were significantly reduced(P<0.01).There was no significant difference in the modification frequency and the number of uprights(P>0.05).Neurons in the VLPO brain region showed pathological damage.The protein and mRNA expression levels of NR1 and CaMK I were decreased(P<0.01)in VLPO brain

关 键 词：摩腹法 揿针 焦虑性失眠 睡眠内稳态系统 腹外侧视前核(VLPO) NMDA CaMKⅡ 大鼠 

分 类 号：R245.31[医药卫生—针灸推拿学]