新疆医科大学第四临床医学院,新疆维吾尔自治区乌鲁木齐市新市区新医路393号,830011
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收稿:2022-01-30,
修回:2022-07-04,
纸质出版:2022-10-02
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闫德祺,张星平,张文慧等.桂枝加龙骨牡蛎汤对肺不藏魄型不寐模型大鼠睡眠情况、血清及脑组织5羟色胺水平的影响[J].中医杂志,2022,63(19):1865-1871.
闫德祺,张星平,张文慧等.桂枝加龙骨牡蛎汤对肺不藏魄型不寐模型大鼠睡眠情况、血清及脑组织5羟色胺水平的影响[J].中医杂志,2022,63(19):1865-1871. DOI: 10.13288/j.11-2166/r.2022.19.013.
目的
2
探讨桂枝加龙骨牡蛎汤对肺不藏魄型不寐的影响及可能作用机制。
方法
2
48只SD大鼠随机分为造模组36只和正常组12只。造模组大鼠采用长平台浅水环境法剥夺大鼠睡眠42天制备肺不藏魄型不寐大鼠模型。造模第15天造模组大鼠随机分为模型组、右佐匹克隆组、桂枝加龙骨牡蛎汤组,每组12只,继续进行造模28天,同时桂枝加龙骨牡蛎汤组灌胃给予桂枝加龙骨牡蛎汤7.6 g/(kg·d),右佐匹克隆组灌胃给予右佐匹克隆片0.105 mg/(kg·d),模型组和正常组灌胃给予0.5 ml/100 g生理盐水。在给药第22-28天统计每日8:00、12:00、16:00、20:00时睡眠大鼠的数量;28天干预结束后第2天进行旷场实验统计大鼠站立次数、运动距离、静止时间,监测并分别统计8:00-20:00期间大鼠非快速眼球运动(NREM)、快速眼球运动(REM)睡眠时间以及清醒时间所占的比例;旷场实验结束后采用液质法检测大鼠血清和脑额叶5-HT含量,采用免疫组化法检测大鼠脑皮质、下丘脑和脑干5-HT表达。
结果
2
与正常组比较,模型组8:00、12:00、16:00时睡眠大鼠数量减少,20:00时睡眠大鼠数量增加,旷场实验站立次数减少,NREM、REM所占比例均下降、清醒时间所占比例增加,血清和脑额叶5-HT含量升高,脑皮质、下丘脑和脑干5-HT表达均升高(
P
<0.05)。与模型组比较,右佐匹克隆组在8:00、12:00、16:00时睡眠大鼠数量增加,旷场实验站立次数增加,REM所占比例升高(
P
<0.05),血清和脑组织5-HT水平差异无统计学意义(
P
>0.05);桂枝加龙骨牡蛎汤组在8:00、16:00时睡眠大鼠数量增加,20:00时减少,旷场实验运动距离缩短,REM所占比例升高,血清和脑额叶5-HT含量下降,脑皮质、下丘脑和脑干5-HT表达均下降(
P
<0.05)。与右佐匹克隆组比较,桂枝加龙骨牡蛎汤组旷场实验大鼠站立次数和运动距离均降低、静止时间增加,REM所占比例升高(
P
<0.05)。
结论
2
桂枝加龙骨牡蛎汤能够改善肺不藏魄型不寐模型大鼠的睡眠情况,恢复血清及脑组织中5-HT水平可能是其作用机制之一。
Objective
2
To investigate the effects of
Guizhi Longgu Muli Decoction
(桂枝加龙骨牡蛎汤, GLMD) on insomnia model rats with the syndrome of lung failing to store corporeal soul, and the underlying mechanism.
Methods
2
Forty-eight SD rats were randomly divided into modeling group (n=36) and normal group (n=12). Rats in the modeling group were raised in the shallow water environment with long platforms in it and were deprived of sleep for 42 days to prepare the insomnia model with lung failing to store corporeal soul syndrome. On the 15th day of modeling, the rats in the modeling group were further randomly divided into model group, eszopiclone group and GLMD group with 12 rats in each group. The modelling was continued for 28 days, and simultaneously 7.6 g/(kg·d) of GLMD and 0.105 mg/(kg·d) of eszopiclone tablets were given by gavage to the mice in the GLMD group and eszopiclone group, respectively, while 0.5 ml/100 g of normal saline by gavage was given to those in the model group and normal group. From the 22nd to the 28th day of the intervention, the number of sleeping rats was counted at 8:00, 12:00, 16:00 and 20:00 everyday with the help of a infrared night vision device. After 28-day intervention, the open field experiment and EEG/EMG monitoring were carried out on the next day to count the standing times, movement distance and resting time, and calculate the the proportion of non-rapid eye movement (NREM), rapid eye movement (REM) sleep time and awake time of the rats during the period from 8:00 to 20:00, respectively. The serum and brain 5-HT was detected by liquid mass spectrometry after the field experiment time, and the 5-HT expression in cerebral cortex, hypothalamus and brainstem was detected by immunohistochemistry.
Results
2
Compared to those in the normal group, the number of sleeping rats at 8:00, 12:00 and 16:00 in the model group decreased, while the number of sleeping rats at 20:00 increased; moreover, the standing times, the NREM and REM sleep time from 8:00 to 20:00 decreased, while the awake time, serum and brain 5-HT, and the expression of 5-HT in cerebral cortex, hypothalamus and brainstem increased (
P
<0.05). Compared to those in the model group, the number of sleeping rats at 8:00, 12:00 and 16:00, the standing times, and REM proportion increased (
P
<0.05), with no significant difference in serum and brain 5-HT level in the eszopiclone group (
P
>0.05); the number of sleeping rats at 8:00, 12:00 and 16:00 and the open field experiment movement distance significantly decreased in the GLMD group, while the REM proportion increased, and serum and brain 5-HT level and the expression of 5-HT in the cerebral cortex, hypothalamus and brainstem decreased (
P
<0.05). GLMD group had decreased standing times and movement distance, increased resting time, and REM proportion than the eszopiclone group (
P
<0.05).
Conclusion
2
GLMD can improve the sleep condition of insomnia model rats with lung failing to store corporeal soul syndrome, and the mechanism may be related to the restore of serum and brain 5-HT level.
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