Influence of drug treatment on sleep in zebra fishes with insomnia and relevant mechanism

HE Dong-fang; ZHANG Li-jun; LIU Mei-yan

doi:10.3969/j.issn.1674-4055.2017.03.20

Abstract：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; 167μg/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) svs. (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 high-dose 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 (allP>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 (allP>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 high-dose subgroup compared with insomnia subgroup (allP<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.