摘要
目的探讨摩腹联合电针透穴法对失眠大鼠生物钟基因Clock、BMAL1、PER1表达及神经递质乙酰胆碱(ACh)、谷氨酸(Glu)含量的影响及可能的作用机制。方法50只SD雄性大鼠随机分为正常组、模型组、摩腹组、电针透穴组和联合组,每组10只。除正常组外,采用腹腔注射对氯苯丙氨酸建立失眠大鼠模型。摩腹组进行腹部按摩,电针透穴组采用平刺法,百会透神庭、三阴交透阴陵泉(双侧)、神门透内关(双侧),电针仪疏密波,频率1Hz/20Hz,联合组摩腹后进行电针透穴治疗,各组均1次/d,连续7d,正常组和模型组不予干预。HE染色观察下丘脑组织神经元细胞形态变化,RT-qPCR检测下丘脑组织Clock、BMAL1、PER1 mRNA表达,免疫组化染色检测下丘脑组织Clock、BMAL1、PER1表达,Western blot检测下丘脑组织Clock、BMAL1、PER1蛋白表达,ELISA检测血清ACh、Glu含量。结果与正常组比较,模型组大鼠入睡潜伏期延长,睡眠持续时间缩短,下丘脑组织细胞结构破坏严重、呈空泡样变化,神经元细胞数量减少,下丘脑组织Clock、BMAL1mRNA和蛋白表达升高,PER1mRNA和蛋白表达降低,血清ACh、Glu含量增加,差异均有统计学意义(P<0.05);与模型组比较,摩腹组、电针透穴组和联合组均能缩短入睡潜伏期,延长睡眠持续时间,改善下丘脑神经元细胞形态,降低下丘脑组织Clock、BMAL1mRNA和蛋白表达,上调PER1mRNA和蛋白表达,减少血清ACh、Glu含量,差异均有统计学意义(P<0.05),以联合组作用最显著(P<0.05)。结论摩腹联合电针透穴法能改善大鼠失眠状况,其机制可能与调控生物钟基因Clock、BMAL1、PER1mRNA和蛋白表达及神经递质含量有关。
Objective To investigate the effects of abdominal massage combined with electroacupuncture point-through-point method on the expressions of circadian clock genes of Clock,BMAL1,PER1 and neurotransmitters contents of ACh and Glu in insomnia rats and its possible mechanism.Methods Totally 50 SD male rats were randomly divided into normal group,model group,abdominal massage group,electroacupuncture point-through-point group and combination group,with 10 rats in each group.Except for the normal group,an insomnia rat model was established by intraperitoneal injection of p-chlorophenylalanine.Abdominal massage group received abdominal massage;electroacupuncture point-through-point group was treated with flat acupuncture,“Baihui”through“Shenting”,“Sanyinjiao”through“Yinlingquan”(bilateral),“Shenmen”through“Neiguan”(bilateral),the electroacupuncture instrument dilatational wave,frequency of 1 Hz/20 Hz was connected;the combination group was treated with electroacupuncture through acupoints after abdominal massage;1 time/d for each treatment group,a total of 7 days.The normal group and model group were not intervened.HE staining was used to observe the pathological changes of neurons in hypothalamic tissue,the expression of Clock,BMAL1 and PER1 mRNA were detected by RT-qPCR,the expressions of Clock,BMAL1 and PER1 in hypothalamic tissue were detected by immunohistochemical staining,the expression of Clock,BMAL1 and PER1 protein in hypothalamic tissue were detected by Western blot,the contents of ACh and Glu in serum were detected by ELISA.Results Compared with the normal group,the model group rats had a longer sleep latency,shorter sleep duration,severe damage to the cellular structure of the hypothalamic tissue,and a vacuolar like change,with a decrease in the number of neuronal cells,the expressions of Clock and BMAL1 mRNA and protein in hypothalamic tissue increased,while the expressions of PER1 mRNA and protein decreased;the contents of serum ACh and Glu increased,with statistical significance(P<0.05).Compared with the model group,the abdominal massage group,electroacupuncture point-through-point group,and combination group could all shorten the sleep latency,prolong sleep duration,and improve the morphology of hypothalamic neurons,reduce the expressions of Clock and BMAL1 mRNA and protein in hypothalamic tissue,up-regulate the expressions of PER1 mRNA and protein,and reduce the contents of serum ACh and Glu,with statistical significance(P<0.05),the combination group showed the most obvious effects(P<0.05).Conclusion Abdominal massage combined with electroacupuncture point-through-point method can improve insomnia.Its mechanism may be related to the regulation of circadian clock genes Clock,BMAL1,PER1 mRNA and protein expression,as well as neurotransmitter content.
作者
张雪枫
邱思淦
燕子悦
古来撒尔·艾克拜尔
卢旭昇
郭蕾
刘俊昌
张星平
ZHANG Xuefeng;QIU Sigan;YAN Ziyue;Gulaisaer Aikebaier;LU Xusheng;GUO Lei;LIU Junchang;ZHANG Xingping(Xinjiang Medical University,Urumqi 830017,China;Xinjiang Medical University Affiliated Hospital of Traditional Chinese Medicine,Urumqi 830000,China;Xinjiang Uygur Autonomous Region Academy of Traditional Chinese Medicine,Urumqi 830000,China)
出处
《中国中医药信息杂志》
CAS
CSCD
2024年第6期95-101,共7页
Chinese Journal of Information on Traditional Chinese Medicine
基金
国家自然科学基金地区基金(81960906、82160873)
新疆维吾尔自治区科技创新团队(天山创新团队)项目(2022TSYCTD0008)。
关键词
摩腹
电针透穴法
失眠
生物钟基因
神经递质
大鼠
abdominal massage
electroacupuncture point-through-point method
insomnia
circadian clock gene
neurotransmitters
rats
