桂枝甘草龙骨牡蛎汤类方对斑马鱼睡眠行为的影响

Effects of Guizhi Gancao Longgu Muli-like decoctions on the sleep behavior of zebrafishes

目的研究桂枝甘草龙骨牡蛎汤类方对斑马鱼睡眠行为的影响。方法 96孔板分为8组,每组12孔,依次分为:空白对照组、褪黑素组、四逆加桂甘龙牡组、柴胡加龙骨牡蛎组、桂枝加龙骨牡蛎组、桂甘龙牡组6个组。将受精后第5天的野生型幼鱼用加样枪逐个加入96孔板中,每孔连幼鱼的体积为300μL,然后根据药物分组每孔用移液枪加入配置好浓度的药物60μL,空白对照组加入等量的含甲基蓝的Holt Buffer培养液,放置在幼鱼观察行为仪装置中观察。用视频跟踪系统记录不同给药组斑马鱼的休息及运动活力指标,同时给予光周期刺激,观察斑马鱼的运动活性。结果与空白对照组比较,褪黑色素组增加了睡眠总时间,减少了总的活动量和觉醒的活动量(P<0.05)。四逆加桂甘龙牡组增加了黑夜睡眠总时间,减少了总的活动量,增加了觉醒的活动量(P<0.05)。柴胡加龙骨牡蛎组、桂枝加龙骨牡蛎组和桂甘龙牡组均增加了睡眠总时间(P<0.05),降低了总的活动量和觉醒的活动量(P<0.05)。相关性分析显示四逆加桂甘龙牡组与桂甘龙牡组、柴胡加龙骨牡蛎组、桂枝加龙骨牡蛎组不具有相同作用神经通路。而桂甘龙牡组、柴胡加龙骨牡蛎组、桂枝加龙骨牡蛎组可能具有相似的神经通路。结论桂枝甘草龙骨牡蛎汤类方对斑马鱼的运动行为有抑制作用,不同配伍对斑马鱼行为节律影响不同,其作用通路亦不尽相同。

Objective To study the effects of Guizhi Gancao Longgu Muli - like decoctions on the zebrafish sleep behavior. Methods The video - tracking system was used to record resting and activity indexes of zebrafishes in different administration groups. At the same time, photoperiodic stimulation was given to observe zebrafish＇s motility. Results Compared with blank group, the total sleep total was significantly increased, as the total activity and waking activity were significantly reduced in positive control group （melatonin） （ P 〈 0. 05 ）. Sini jia guigan longmu prescriptions significantly increased the total sleep in nighttime, reduced the total activity and increased the waking activity （ P 〈 0. 05 ）. Chaihu jia longgu muli prescriptions, Guizhi jia longgu muli prescriptions and Guigan longmu prescriptions all significantly increased the total sleep total, and reduced the total activity and waking activity （ P 〈 0. 05 ）. Correlation analysis showed that Sini jia guigan longmu decoction did not have the same biological targets with Guigan longmu decoction, Chaihu jia longgu muli decoction or Guizhi jia longgu muli decoction. Guigan longmu decoction, Chaihu jia longgu muli decoction and Guizhi jia longgu muli decoction may have similar biological targets. Conclusion Guizhi Gancao Longgu Muli - like decoctions have inhibitory effect on the zebrafish＇s activity. Different compatibility has different effects on the zebrafish＇s rhythm, and their biological targets are also different.