久坐湿地法建立肾不藏志不寐大鼠模型的研究

Study on establishment of a rat model of insomnia with kidney not being able to store zhi by long time sitting on the wetland

目的:为依据单一主诉确立的中医不寐五神分型诊断法的研究建立一种肾不藏志不寐大鼠模型。方法:24只雄性SD大鼠随机分为正常组和模型组,每组12只。正常组大鼠正常饲养,不给予干预。模型组大鼠于中午换到装有10 cm厚的4℃湿沙的大鼠笼内,夜幕降临时放入15~18℃水中后立即取出放回正常笼中,实验21 d。以大鼠2导脑电图/肌电图为主评价大鼠睡眠情况,以体质量、戊巴比妥钠睡眠诱导实验、水迷宫实验、血清褪黑素(MT)水平和脑干、中段脑和肾皮质褪黑素相关受体(GPR50)水平为辅探讨不寐引起的功能和节律分子水平的变化。结果:与正常组比较,模型组一般情况较差,体质量显著降低(P<0.05)。模型大鼠白天各期睡眠时间显著缩短(P<0.05),以非快速动眼睡眠(NREM)(3,4)期和快速眼动睡眠(REM)减少为主;夜间各期睡眠时间显著增加(P<0.05),以NREM(1,2)期和低电压REM睡眠增加为主;戊巴比妥钠诱导的睡眠潜伏期显著延长、睡眠时间显著缩短(P<0.05);水迷宫实验提示逃避于平台上的潜伏期显著增加(P<0.05);穿台次数显著减少(P<0.05);血清MT水平显著降低(P<0.05),除听觉区外的其他受试脑区GPR50水平显著升高(P<0.05);肾皮质GPR50显著降低(P<0.05)。结论:采用大鼠睡眠后半期“久坐湿地,强力入水”法可以建立以早醒为特点的肾不藏志不寐大鼠模型,为研究依据单一主诉确立的中医不寐五神分型诊断法提供了动物模型。

Objective:To establish a rat model of insomnia with kidney not being able to store zhi for the study of diagnostic method of insomnia in traditional Chinese medicine five shen by unique complaint.Methods:Twenty-four male Sprague Dawley(SD)rats were randomly distributed into normal group and model group(n=12).The rats in the normal group were fed normally without intervention.The rats in the model group were transferred to the cage with 10 cm thick 4℃wet sand after noon,and put into 15~18℃water before returning to the normal cage after nightfall everyday in the 21 days.2EEG/EMG was used to evaluate the sleep of the rats.Body weight,pentobarbital sodium induced sleep test,water maze experiment,melatonin(MT)and GPR50 were used to elucidate some function and molecular level changes.Results:Compared with the normal group,the general condition of the model group was poor and the body weight decreased significantly(P<0.05).The sleep time of model rats in each stage of daytime sleep was significantly shortened(P<0.05),mainly in non-rapid eye movement sleep(NREM)(3,4)and rapid eye movement sleep(REM)sleep;the sleep time at night increased significantly(P<0.05),especially in NREM(1,2)and lower voltage REM sleep;Pentobarbital sodium induced prolonged sleep latency(P<0.05)and shortened sleep time(P<0.05);the night water maze test showed that the latency of escaping onto the platform increased significantly(P<0.05);the times of crossing the platform decreased significantly(P<0.05);the level of serum MT decreased(P<0.05),and the level of brain GPR50 increased(P<0.05);GPR50 in renal cortex decreased(P<0.05).Conclusion:The method of‘long time sitting on the wetland and entering the water quickly’in the second half of rat sleep can establish the insomnic rat model of‘kidney being not able store zhi’,and provide an animal model for studying the insomnia classification by Five shen in traditional Chinese medicine.