摘要
目的观察药物处理对失眠症斑马鱼睡眠的改善作用,并探讨银杏叶滴丸(GBP)对γ-氨基丁酸A型受体α1亚单位(GABRA1)和褪黑素受体1类亚型(MTNR1)mRNA表达的影响。方法采用咖啡因诱导活体斑马鱼建立失眠模型,分别水溶给予GBP(低剂量组56μg/mL、中剂量组167μg/mL、高剂量组500μg/mL)、金纳多(Ginaton,低剂量组56μg/mL、中剂量组167μg/mL、高剂量组500μg/mL)和氯化锂(阳性对照组,500μM),同时设置正常对照组和失眠组,应用行为分析仪定量斑马鱼处于失眠状态的时间,评价GBP对斑马鱼睡眠的改善作用。另取失眠模型斑马鱼,水溶给予GBP56、167、500μg/mL,同时设置正常对照组和失眠组,采用RT-PCR检测GABRA1和MTNR1 mRNA的相对表达量。结果在观察的1 h(3600 s)内,失眠组斑马鱼失眠时间多于正常对照组,(1598±277)s vs.(1240±129)s,差异有统计学意义(P<0.01)。阳性对照组斑马鱼失眠时间为(1368±170)s,少于失眠组,差异有统计学意义(P<0.05)。低剂量、中剂量、高剂量的GBP对睡眠的改善作用分别为72%、95%和97%。低剂量、中剂量、高剂量的Ginaton对睡眠的改善作用分别为78%、80%和95%。与正常对照组比较,GBP低剂量组GABRA1 mRNA相对表达量降低,差异有统计学意义(P<0.01);失眠组、GBP中剂量组和高剂量组无明显变化,差异无统计学意义(P均>0.05);与失眠组比较,GBP低剂量组GABRA1 mRNA相对表达量降低,差异有统计学意义(P<0.01)。MTNR1 mRNA相对表达量:与正常对照组比较,失眠组、GBP低剂量组和高剂量组降低,差异有统计学意义(P均<0.01),GBP中剂量组无明显变化,差异无统计学意义(P>0.05);与失眠组比较,GBP低剂量组、中剂量组和高剂量组均升高,差异有统计学意义(P均<0.01)。结论低、中、高浓度银杏叶滴丸和金纳多对失眠症斑马鱼睡眠均有显著改善作用;银杏叶滴丸改善睡眠的可能机制为抑制GABRA1的表达和改善MTNR1的表达。
Objective To observe the relief effect of drug treatment on sleep in zebra fishes with insomnia, and discuss the influence of Ginkgo Biloba Pills (GBP) on mRNA expressions of γ-aminobutyric acid type A receptor-1 (GABRA1) and melatonin receptor-1 (MTNR1). Methods The insomnia model was established by caffeine in living zebra fishes, and zebra fishes were respectively given GBP (56 μg/mL, GBP low-dose group; 167 μg/mL, GBP mid-dose group; 500 μg/mL, GBP high-dose group), Ginaton (56 μg/mL, Ginaton low-dose group;167 μg/mL, Ginaton mid-dose group; 500 μg/mL, Ginaton high-dose group), and lithium chloride (500 μM, positive control group), and at the same time normal control group and insomnia group was set up. The duration of zebra fishes being insomnia condition was quantified by using behavior analyzer, and relief effect of GBP on zebra fishes’ sleep was reviewed. In addition zebra fishes with insomnia were selected and given GBP [56 μg/mL (GBP low-dose subgroup), 167 μg/mL (GBP mid-dose subgroup) and 500 μg/mL (GBP high-dose subgroup)],and at the same time normal control subgroup and insomnia subgroup was set up for detecting the relative mRNA expressions of GABRA1和MTNR1 by using RT-PCR. Results The insomnia duration was longer in insomnia group than that in normal control group [(1598 ±277) s vs. (1240 ±129) s, P<0.01] within observation time for 1 h (3600 s). The insomnia duration was shorter in positive control group (1368±170) s than that in insomnia group (P<0.05). The meliorating effect of GBP on sleep reached, respectively, by 72% in GBP low-dose group, by 95% in GBP mid-dose group and by 97% in GBP high-dose group. The meliorating effect of Ginaton on sleep reached,respectively, by 78% in Ginaton low-dose group, by 80% in Ginaton mid-dose group and by 95% in Ginaton highdose group. The relative expression of GABRA1 mRNA decreased in GBP low-dose subgroup compared with normal control subgroup (P<0.01), had no significant changes in insomnia subgroup, GBP mid-dose subgroup and GBP high-dose subgroup (all P>0.05), and decreased in GBP low-dose subgroup compared with insomnia subgroup (P<0.01). The relative expression of MTNR1 mRNA decreased in insomnia subgroup, GBP low-dose subgroup and GBP high-dose subgroup compared with normal control subgroup (all P>0.01), had no significant changes in GBP mid-dose subgroup (P>0.05), and increased in GBP low-dose subgroup, GBP mid-dose subgroup and GBP highdose subgroup compared with insomnia subgroup (all P<0.01). Conclusion Low-dose, mid-dose and high-dose GBP and Ginaton all have significant meliorating effect on sleep in zebra fishes with insomnia, and the possible mechanism may be the inhibition of GABRA1 expression and melioration of MTNR1 expression.
作者
何东方
张丽军
刘梅颜
HE Dong-fang;ZHANG Li-jun;LIU Mei-yan(Department of Cardiology, Beijing Anzhen Hospital, Capital University of Medical Sciences, Beijing 100029, China)
出处
《中国循证心血管医学杂志》
2017年第3期326-330,334,共6页
Chinese Journal of Evidence-Based Cardiovascular Medicine
关键词
斑马鱼
失眠
抑郁
γ-氨基丁酸A型受体
褪黑素受体
Zebra fishes
Insomnia
Depression
γ-Aminobutyric acid type A receptor
Melatonin receptor
