| Objective:To sort out the ancient Chinese books related to dementia and insomnia,clarify the comorbid mechanism of the two in Chinese medicine,and clarify the traditional Chinese medicine prescriptions for preventing and treating dementia and insomnia.Based on the research foundation of the research group,6-month-old and 9-month-old APP/PS1 double-transgenic dementia mice were selected,and 6-month-old dementia mice were given Suanzaoren Decoction to explore Suanzaoren decoction through behavioral and molecular biological experimental techniques.The role of dementia with sleep disorders and the mechanism of action,so as to provide reliable theoretical basis and experimental basis for prevention and treatment of dementia and insomnia.Method:1.Theoretical discussion:The literature on ancient Chinese medicine related to dementia and insomnia was consulted,the relevance of dementia and insomnia was clarified from the perspective of traditional Chinese medicine,and the comorbid mechanism of dementia and insomnia was elaborated,thus clarifying the rational prescriptions for preventing and treating dementia and insomnia.2.Experimental study:The experiment used APP/PS1 double transgenic mice and Suanzaoren decoction as the research object to explore the effect and mechanism of Suanzaoren Decoction in treating AD sleep disorders.The 6-month-old and9-month-old APP/PS1 double transgenes were selected for the experiment.The mice were observed for sleep rhythms of 6-month-old and 9-month-old APP/PS1 double transgenic mice.The 6-month-old APP/PS1 transgenic mouse was given Suanzaoren Decoction.The effect of Suanzaoren decoction on sleep rhythm of 6-month-old dementia mice was observed through independent activity experiments;Morris water maze test was used to detect the effect of Suanzaoren decoction on 6-month-old dementia mice;the application of HE staining,Nissl's staining,and immunity Histochemical staining was used to observe the histopathological changes in hippocampus of6-month-old dementia mice by histochemical staining;the effect of Suanzaoren decoction on serum IL-1?and TNF-?in 6-month old dementia mice was detected by ELISA;RealTime-PCR was used.The effect of rhythm mRNA expression in hypothalamus of 6-month-old dementia mice was detected.The expression of A?1-40,A?1-42,GFAP and IBA-1 in hippocampus of 6-month-old dementia mice was determined by Western Blot.Results:1.Sleep rhythm experiment resultsCompared with the blank group,there was no significant difference in activity time of the dark environment between the mice of the 6 months old group and the blank group?P>0.05?,and the total activity time and activity time of the light environment increased significantly?P<0.01?.In 9-month-old mice,compared with the blank group,the activity time of dark environment,activity time of light environment,and total activity time of mice in APP/PS1group increased to varying degrees?P<0.01?.2.Behavioral experiment resultsWater maze test results:Compared with the model group,the latency of the upper platform of each group of mice was shortened?P<0.01,P<0.05?,and the total swimming distance decreased?P<0.01,P<0.05?;compared with the donepezil group,The upper platform latent period and total swimming distance of mice in high-dose Suanzaoren decoction groups increased,but the difference was not statistically significant?P>0.05?.The total swimming distance of mice in the low-dose group of Suanzaoren decoction increased?P<0.05?.The latency of the upper platform also increased,but the difference was not statistically significant?P>0.05?.Compared with the model group,the times of crossing platform and target quadrant of the mice in the blank group,donepezil group,and high-dose Suanzaotang group increased?P<0.01?,and the time to reach the original platform on the first time was shortened?P<0.01?.Suanzaoren Decoction The number of mice crossing the platform in the low-dose group increased?P<0.01?,the time to reach the original platform reached the first time?P<0.01?,and the target quadrant time increased,but the difference was not statistically significant;compared with the donepezil group,the Suanzaoren Tanggao The number of times that the dose group traversed the platform decreased?P<0.05?.The time of the target quadrant and the time of the first arrival at the original platform decreased,but the difference was not statistically significant?P>0.05?.The target quadrant time of the low-dose group of Suanzaoren decoction Decreased?P<0.05?,decreased number of crossing platforms,and increased time to first platform arrival,but the difference was not statistically significant?P>0.05?.The step-down test results:compared with the model group,the number of errors in the blank group mice was significantly reduced?P<0.01?,and the latency period of the platform latency was shortened?P<0.05?,the donepezil group,high and low doses of Suanzaoren Decoction,and low doses of Suanzaoren Decoction.The number of rat errors increased and the latent period latency of the platform was shortened,but the difference was not statistically significant?P>0.05?.Compared with the donepezil group,the number of error times and the latent period of latent period of the low-dose Suanzaoren Decoction and Suanzaoren Decoction mice were changed.,but the difference was not statistically significant.Sleep rhythm test results:Compared with the model group,dark activity time,total activity time,and light activity time of the mice in other groups were reduced at the age of 6 months?P<0.01,P<0.05?.Compared with the donepezil group,the total activity time and light activity time of the high-dose Suanzaoren Decoction group decreased?P<0.01?,and the total activity time and light activity time of the Suanzaoren Decoction group decreased,but there was no statistical significance?P>0.05?.3.ELISA test resultsCompared with the model group,TNF-?and IL-1?in the serum of the blank group were significantly decreased?P<0.01?.The levels of TNF-?and IL-1?in the serum of the mice in the blank group were decreased to different degrees?P<0.01,P<0.05?.4.Real Time-PCR experiment resultsCompared with the model group,the expression of Per1 mRNA,Cry1mRNA,Bmall mRNA and Clock mRNA were up-regulated in the other groups.Compared with Donepezil group,the expression of Per1 mRNA,Bmall mRNA,and Clock mRNA were down-regulated in high-dose Suanzaoren decoction group,but Cry1 mRNA and Clock mRNA were up-regulated.Perl mRNA,Cry1 mRNA,Bmall mRNA,and Clock m RNA were down-regulated in Suanzaoren Decoction low-dose group.5.Pathological experiment resultsHE staining results:In the blank control group?C57/BL6J?mice,the vertebral body cells in the DG region of the hippocampus were well organized and well-distributed,with uniform staining,complete structure,tight cell connections,and obvious nucleoli.In the APP/PS 1 mouse hippocampal DG area,the vertebral body cells were arranged neatly,the morphology was intact,the nucleolus were unclear,the nucleus was deeply stained,the phenomenon of nuclear pyknosis occurred,and the intercellular space of the local neurons increased.The model group was small.In the hippocampus of rats?APP/PS1?,the arrangement of nerve vertebral cells in the DG region is still neat,some intercellular spaces between neurons increase,most neuronal nuclei show a pyknosis,deep cytoplasm staining,and the nuclear membrane boundary is not Clearly,necrotic neurons were visible,and in the donepezil group,some neurons showed a nucleus pyknosis,some deep staining of the cytoplasm was evident,and the cell arrangement was still neat,but the intercellular space between neurons increased significantly.In the low-dosage of Suanzaoren decoction,the neurons in the hippocampus were arranged neatly and the number of neurons was slightly increased.Occasionally,the nucleus was pyknosis and the nuclei were increased.The high-dose Suanzaoren group was more obvious.Nissl staining results:In the blank group,the neurons in the DG region of the hippocampus were arranged in a regular pattern,with clear and light Nissl bodies in the cells,but some neurons had pyknosis and dark color.In the model group,the neurons in the DG region of the hippocampus of mice were disordered,with many darker nuclei,darker intracellular Nissl staining,and even pyknosis.After administration,the neurons in the DG neurons of the hippocampus in the doneepez group,the high-dose group of the Suanzaoren,and the low-dose Suanzaoren groups were significantly increased compared with the model group,and arranged more neatly,and the cell layer was rich,but there were also more Nyquist neurons Body,in which the high and low doses of Suanzaoren's Nissl body's pyknosis and deep stain improved most obviously.Immunohistochemical results:Immunohistochemistry results of A?1-40and A?1-42 showed that the blank group had intact hippocampal cells,neat cells with normal morphology,and light yellow-brown granules on the cell membrane and cytoplasm.Compared with the blank group,the mouse hippocampal cells in the model group showed brown staining and incomplete cells.The rest of the groups were stained with light brown particles.GFAP immunohistochemistry results showed that compared with the blank group,the number of stained cells in the model group increased significantly,and the staining deepened.Compared with the model group,cells in each administration group stained more deeply,especially in the high-dose group.IBA1 immunohistochemistry results showed that compared with the blank group,the IBA1 positive cell body in the model group was darkly stained and the cell body was slightly enlarged.Compared with the model group,the staining of the IBA1 positive cells in the other groups was lighter.Western Blot experiment resultsContent of A?1-40 and A?1-42 protein in hippocampus:Compared with the model group,the relative expression of A?1-40-40 and A?1-42-42 protein in the hippocampus of each group was significantly decreased?P<0.01,P<0.05?.Compared with the donepezil group,the relative expression of A?1-40 protein in the hippocampus of the high-dose and low-dose groups of Suanzaoren Decoction significantly increased?P<0.01?,and the expression of A?1-42increased in the Suanzaoren Decoction low-dose group?P<0.05?.The expression of A?1-42 in the high-dose Suanzaoren Decoction group also increased,but the difference was not statistically significant?P>0.05?The content of GFAP and IBA1 protein in hippocampus:Compared with the model group,the expression of GFAP and IBA1 decreased in all groups?P<0.05,P<0.01?.Compared with the donepezil group,the expression of GFAP and IBA1 in the high dose group of Suanzaoren Decoction significantly decreased?P<0.01?,the expression of GFAP in the low dose group of Suanzaoren also decreased?P<0.05?,while the expression of IBA1 decreased in the Suanzaoren low dose group.However,the difference was not statistically significant?P>0.05?.Conclusion:1.Dementia and insomnia are related in many aspects such as etiology,pathogenesis,pathological features,site of onset,and prescriptions for treatment.Deficiency of the heart and brain is a common pathogenesis of dementia and insomnia.Therefore,dementia with sleep disorders can be divided from Heart,brain treatment,or heart and brain Tongzhi.2.The 6-month-old APP/PS1 double transgenic mice have obvious sleep rhythm disturbances and can be used as an animal model for studying AD sleep disorders.3.Suanzaoren soup can improve the learning and memory of APP/PS1double transgenic mice and improve sleep disorders.4.The mechanism of improving and improving the learning and memory and sleep disturbance of APP/PS1 double transgenic mice by sojazao decoction may be related to the reduction of inflammatory reaction,the regulation of circadian clock genes and the protection of neurons. |
PDF Full Text Request