| Objective:Through the induction and arrangement of ancient and modern literature,this study theoretically discussed the origin,etiology and pathogenesis of senile insomnia and the basis of Guben-Jiannao liquid in the treatment of senile insomnia,at the same time by observing the thrift brain fluid behavior of senile insomnia model rats,EEG electromyography and the influence of relevant indicators protein,can clear up brain fluid based on GABA conduction system for inhibitory protective effect and mechanism of senile insomnia for thrift brain fluid clinical use provides the experimental basis and theoretical guidance.Methods:60 healthy SD rats were randomly divided into 4 groups:normal group?n=15?,model group?n=15?,estazolam group?n=15?and Guben-Jiannao liquid group?n=15?.after adaptive feeding for 1w,build module?model group,Guben-Jiannao liquid group,estazolam group?was injected subcutaneously with D-galactose 200mg.kg-1.d-1 on the back of neck for 8 weeks,which was an aging model,and the normal group was subcutaneously injected with the same dose of normal saline at the same time.Then Guben-Jiannao group,estazolam and model group were put into the self-made sleep deprivation box.Sleep deprivation was carried out for 18 hours from 07:00 every day for 21 days.All behaviors of model rats in each group were detected and analyzed by HomecageScan3.0 real-time monitoring system,and the observation time was 24hours to detect the model.After the molding is establishment,5 experimental rats in each group were implanted with electrodes.On the first day after operation,the estazolam group was given a dose of 0.09mg.kg-1.d-1,and the GubenJiannao liquid group was given a dose of 16g.Kg-1.d-1.The normal group,the model group and the estazolam group were treated with the same dose of normal saline as the experimental control group.All the animals were given the same dose of normal saline at 18:00every night for 2 weeks.EEG and EMG were recorded on the first day after administration for 24 hours.The hypothalamus morphology was observed by H.E.staining,and the expression of GABAAR 1,GABAAR 2 and NKCC1 were detected by immunohistochemistry and western blot.Results:HomeCage monitoring showed that rats in the model group were more likely to be inactive and less active,with significantly lower activity frequency than those in the normal group,the hair was dry and dull,more sleep awakenations,significantly increased drinking water,significantly decreased sleep duration?P<0.05?,more sleep awakenations and reversed circadian rhythm,indicating successful model replication.Compared with the normal group,the cumulative amount of arousal in the model group increased significantly during the day?P<0.05?,the number of awakenings increased significantly?P<0.05?,the cumulative amount of REM sleep decreased significantly?P<0.05?,and the cumulative amount of NREM sleep decreased significantly at night?P<0.05?.There was no significant difference in sleep accumulation during the day in the estazolam group,with a significant increase in sleep duration?P<0.05?,a significant decrease in REM sleep accumulation?P<0.05?,and a significant increase in NREM sleep accumulation at night?P<0.05?.Compared with the normal group,there was no significant difference.The H.E.staining results showed that the rats in the treatment group had basically normal cell distribution and less nuclear atrophy,indicating that eszolam and Guben-Jiannao liquid could improve the cellular aberration of hypothalamus..Immunohistochemistry and Western blotting showed that the protein expression of GABAAR?1 and GABAAR?2 increased and the protein expression of NKCC1 decreased in Guben-Jiannao liquid group.Conclusion:this suggests that GubenJiannao liquid can effectively improve the abnormal state of hypothalamic cells by up-regulating GABAAR?1?GABAAR?2 and reducing NKCC1,it can strengthen the binding of GABA and its receptor,and improve the insomnia of aged rats. |
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