Regulation of Tuina on Sleep Deprivation via Adenosine and A2A Receptor within Basal Forebrain in Mice
Objective:To reveal the effect of Tuina on sleep deprivation and the influence of adenosine and A2A receptor within.basal forebrain.Methods:Sixty-six C57BL/6J mice were randomly divided into blank group,model group and Tuina group.After the sleep deprivation model was prepared by sleep deprivation device for 4 days and Tuina treatment for 6 days,the mice were observed for gen-eral conditions and daily activities.Electroencephalogram and electromyogram signals were recorded through ethology.The mice were analyzed for Wake,non-rapid eye movement(NREM),rapid eye movement(REM)sleep phases and basal forebrain neural activity.ELISA was used to calculate the adenosine content in basal forebrain.The fiber photometry was used to observe the dynamic signals of astrocyte-released adenosine in basal forebrain.Western Blot was utilized to measure the protein expression of adenosine A2A receptor in basal forebrain.Results:①Tuina mostly improve the behavioral outlook and daily activity in sleep-deprived mice.First,the hair color and mental phenotype of the sleep-deprived mice were recovered after Tuina treatment.Second,the daily activity of the Tuina group was significantly higher than that of the model group on the 4th-6th day(P＜0.01),and the difference between the activity on the 6th day and that of the blank group was not statistically significant(P＞0.05).②Tuina probably improve the somnolence phenomenon in-duced by sleep deprivation and the neural activity of basal forebrain.The NREM phase decreased significantly after Tuina treatment(P＜0.01)while the Wake phase was significantly increased(P＜0.01),and the 8 and θ wave activities in basal forebrain was de-creased.③Tuina possibly regulate adenosine and A2A receptor of basal forebrain in sleep-deprived mice.The content of adenosine in sleep-deprived mice basal forebrain decreased significantly after Tuina(P＜0.01).Meanwhile,the astrocyte-released adenosine and the protein expression of A2A receptor were both decreased after Tuina-therapy(P＜0.05).Conclusion:(1)Tuina increases daily activity and reduces basal forebrain δ and θ wave activity in sleep deprived mouse,improving sleep deprivation-induced drowsiness.(2)Tuina can reduce the content of basal forebrain adenosine and the protein expression of A2A receptor,and lower the astrocyte-re-leased adenosine of basal forebrain in sleep-deprived mice,which may be involved in improving the awakening process after sleep dep-rivation.
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