| Objective To explore the effect and mechanism of Root-Securing and Brain-Fortifying Liquid(RSBFL)in the treatment of aging insomnia.Methods Seventy SPF male Sprague-Dawley rats were randomly divided into aging control group(D-gal+LPC),aging insomnia group(D-gal+SD),and estazolam group(D-gal+SD+Estazolam).Guben Jiannao Liquid group(D-gal+SD+RSBFL).All rats were subcutaneously injected with 200 mg/kg/d of D-gal 8w on the back of the neck,which was an aging model.At 07:00 every day,the aging control group was placed in the environmental control box,and the other three groups were placed in the water platform environmental sleep deprivation box for 18 hours and 21 days in a row.This is the aging insomnia model.At this time,Home Cage Scan 3.0 was used to monitor the sleep-wake behavior of the rats in the aging control group and the aging insomnia group for one week,which was to observe the characteristics of insomnia symptoms in the aging insomnia model rats.At the same time,the estazolam group was given estazolam liquid by gavage at the standard of 0.09 mg/kg/d,the Gubenjiannao liquid group was given the standard of 6.75 g/kg/d by gavage with Gubenjiannao liquid,and the aging control.The group and the aging insomnia group were given the same volume of double distilled water as the estazolam group every day.All animals were dosed once every night at 19:00for 1w.After that,Home Cage Scan 3.0 was used to monitor the sleep-wake behavior of the rats in each group,Nissl staining was used to observe the changes of neurons and Nissl bodies in the hypothalamus of the rats in each group,and Elisa was used to detect the content of 5-HT(5-hydroxytryptamine,5-HT)and DA(dopamine,DA)in the hypothalamus of the rats in each group.Western blot was used to detect the expression levels of 5-hydroxytryptamine 1A receptor(5-HT1A)and dopamine receptor D2(DRD2)in the hypothalamus of rats in each group.5-HT1 A mRNA and DRD2 mRNA content in murine hypothalamus.Results(1)Home Cage Scan 3.0 to monitor the characteristics of insomnia in aging insomnia model rats Results: Compared with the rats in the D-gal+LPC group,the daytime sleep time,the number of sleep times and the sleep time of the rats in the D-gal+SD group were significantly decreased,and they were prone to premature death.wake up;sleep time at night and one sleep time also decreased significantly,the number of sleep at night did not change significantly,and sleep in the second half of the night was unstable;sleep time within 24 hours a day decreased significantly,and the time required for one awakening process during the day increased sharply,indicating that the aging insomnia model is successful.(2)Home Cage Scan 3.0monitors the sleep-wake results of rats in each group: Compared with the rats in the D-gal+SD group,the rats in the D-gal+SD+Estazolam group had significant sleep times during the day,sleep time during the day and sleep time at night.There were no significant changes in daytime sleep time,night sleep time,number of night sleep times,and total sleep time in a day.Compared with the rats in the D-gal+SD group,the rats in the D-gal+SD+RSBFL group not only significantly increased the sleep time,sleep times and once sleep time during the day,but also increased the sleep time,sleep times and once sleep time at night.The sleep time was significantly increased,and the sleep time of the rats in the D-gal+SD+RSBFL group also increased significantly within one day.(3)Nissl staining results: Hypothalamus of D-gal+LPC group: the neuron cells have good morphology,compact arrangement,complete cell structure,round or oval nuclei,clear nucleoli,and abundant Nissl in the cytoplasm.Hypothalamus of rats in D-gal+SD group: cells were loosely arranged,some neuronal cell bodies were shrunken,Nissl bodies in the cytoplasm were reduced,and the staining was darker.Compared with the D-gal+SD group,the morphology and number of Nissl bodies in the hypothalamus of the D-gal+SD+Estazolam group and the hypothalamus of the D-gal+SD+RSBFL group were improved.(4)Elisa,WB and PCR results: Compared with rats in D-gal+LPC group,the expressions of 5-HT,5-HT1 A,DA and DRD2 in the hypothalamus of rats in D-gal+SD group were significantly increased,and 5-HT1 Am RN was significantly increased.and DRD2 mRNA expression decreased.Compared with the rats in the D-gal+SD group,the expressions of 5-HT,5-HT1 A,DA and DRD2 in the hypothalamus of the rats in the D-gal+SD+Estazolam group were significantly decreased,and the 5-HT1 A mRNA and DRD2 mRNA were increased;The expressions of 5-HT,5-HT1 A,DA,and DRD2 in the hypothalamus of rats in the D-gal+SD+RSBFL group were also significantly decreased.The mRNA of 5-HT1 A did not change,but the mRNA of DRD2 increased.Conclusion(1)Guben Jiannao Method can significantly improve the insomnia symptoms and pathological changes of hypothalamic neurons in aging insomnia rats,restore their neuron morphology,and increase the number of Nissl bodies.(2)The mechanism of Guben Jiannao Method’s significant improvement of aging insomnia may be related to the content of 5-HT and DA in the hypothalamus,which can down-regulate the expression of 5-HT and its receptor 5-HT1 A,and down-regulate the content of DA and its receptor DRD2,increased DRD2 mRNA. |
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