| Objective: To observe the effect of Gubenjiannao anshen recipe on sleep,learning and memory of senile insomnia rats,which induced by D-galactose and PCPA,and to explore its mechanism by the effect of Gubenjiannao anshen recipe on Circadian Rhythm and Circadian Clock Genes expression.Methods: Sixty rats were divided into aging group,model group,solid high group,solid low group and Western medicine group randomly,12 rats in each group,PCPA was injected intraperitoneally into the rats of other groups except aging group for 7 days,and normal saline was given to the aging group and model group.The sleep time and circadian rhythm of Rats were monitored by Home Cage Scan,the learning and memory ability of rats were measured by Morris water maze,and the m RNA expression levels of Clock,Bmal1,Per1,Per2,Per3,Cry1 and Cry2 in the hippocampus of rats were detected by RT-PCR,the expression of Clock and Bmal1 protein in hippocampus was detected by IHC.HE staining and Nissl staining were used to observe the changes of hippocampal histomorphology and neuronal Nissl bodies.Results: 1.The results of Home Cage Scan showed that compared with the aging group,the sleep time in the model group was reduced significantly（P<0.05）,the Circadian Rhythm disappeared,the sleep rhythm recovered,the nocturnal activity and the daytime sleep time increased significantly（P<0.05）.2.The results of Morris Water Maze Test showed that the time of finding a platform in each group was shortened with the training days,and there was no significant difference in the escape latency between each group and the Aging Group（P>0.05）,the learning efficiency of the high dose group was significantly increased（P<0.05）.The escape latency decreased significantly in the high-dose Group（P<0.05）and the Times of escape increased significantly（P<0.05）.3.The expression of Clock and Bmal1 was significantly decreased（P<0.05）,the expression of Per1,Per2 and Per3 were significantly increased（P<0.05）in the model group compared with the aging group,the expression of Clock and Bmal1 in the High Dose Group of Guben Jiannao Anshen Fang was significantly increased（P<0.05）,and that of Per1 and Per2 were significantly decreased（P<0.05）.It is suggested that Gubenjiannaoanshen decoction may regulate the circadian rhythm of aging and insomnia rats by regulating the Clock,Bmal1,Per1 and Per2 genes.4.The results of IHC showed that the positive expression of Clock and Bmal1 in the hippocampus of rats in the model group was obviously decreased,the arrangement was loose and disorderly,the color was light,and the expression of positive granules was obviously increased after treatment with Guben Jiannao Anshen decoction,the arrangement was close and concentrated,and the shape was regular,darkening of color.5.HE staining and Nissl staining showed that in the aging group,the cell membrane was intact,the nucleus was located in the middle of the cell body,and the nucleoli were obvious;The number of Nissl bodies decreased obviously,and some of Nissl bodies dissolved.After treatment with Guben Jiannao Anshen decoction,the hippocampal cells arranged closely and tidily,the structure was clear,the cell injury recovered remarkably,and a large number of Nissl bodies were found.Conclusion: Insomnia is closely related to the decline of learning and memory ability,Guben Jiannao Anshen decoction has the effects of improving sleep and regulating circadian rhythm in aging insomnia rats induced by injection of D-galactose and PCPA,It can effectively improve the learning and memory function of aging and insomnia rats,and the mechanism may be related to the regulation of biological clock genes. |
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