| Objective The relationship between mind and sleep is discussed on the basis of re-understanding of the relationship between mind,spirit and both from the perspective of traditional Chinese medicine,and re-combing the etiology and pathogenesis of insomnia in traditional Chinese medicine,as well as the treatment of prescription and prescription.On the other hand,from the perspective of immunology to understand the relationship between sleep and inflammatory response and apoptosis.The anti-inflammatory response and anti-apoptosis effects of Yangxin Anshen method on insomnia rats were observed.This research is divided into two parts: theoretical research and experimental research.The aim is to lay a theoretical foundation and reliable experimental basis for the application of the method of nourishing the mind and tranquilizing the mind for insomnia.Method1.Discussion of theory: by consulting and sorting out the relevant literature,to rearrange the understanding of heart and god in the theory of traditional chinese medicine,to summarize the relationship between mind,god and sleep,to study the influence of mind and mind on human sleep,and to explore the application of tranquilizing mind in insomnia.On the other hand,collect data to understand the relationship between sleep and inflammatory response,apoptosis from the perspective of immunology.2.Experimental study: 60 male SD rats were randomly divided into Control group and Model group.Among them,there were 10 in the control group and 50 in the model group.The model was made by intraperitoneal injection of PCPA,the injection dose was350mg·kg-1,and the model was continuously made for 3 days.After modeling,40 of them were selected and divided into model group(isovolume saline)in order,and Tianwang Buxin Dan plus reduced,medium and high dose groups(8.8,17.6,35.2g·kg-1·D-1),10 in each group.Morris water maze method was used to test its learning and memory ability;immunofluorescence staining was used to detect the number and morphology of rat hippocampal neurons;TUNEL staining was used to detect rat hippocampal neuron apoptosis;RT-qPCR was used to observe Changes in gene levels of rat hippocampal inflammatory factor IL-21 and apoptosis regulators Fas / Fas L,Caspase-8,and Bcl-2;Western Blot method was used to detect rat hippocampal inflammatory factor IL-21 and apoptosis regulator Fas/ Fas L,Caspase-8,Bcl-2 protein expression levels.Results1.Model evaluation: Before modeling,the rats in each group were active,good in activity,regular in sleep,and shiny in fur.In the initial stage of modeling,some rats showed excessive excitement,dull hair,white or pale yellow claw nails;in the late stage of modeling,the excitability of the model rats decreased,and their spirits became debilitating.The claw nails are whitish in color,the hair is messy,and the reaction is slow.The control group was in good condition from beginning to end,with flexible activities and normal sleep and food intake.The model was evaluated using the righting reflex method.Intraperitoneal injection of sodium pentobarbital solution 50 mg·kg-1was used to record the latency to sleep and the duration of sleep within 15 minutes.Compared with the control group,the sleep maintenance time of the rats in the model group was significantly shortened,and the latency to fall asleep was significantly prolonged(P<0.01),suggesting that the modeling was successful.2.Morris water maze experiment: the platform incubation period and the total swimming distance of the rats in the model group were significantly longer than those in the control group(P<0.01).After treatment with low,medium and high concentrations of Tianwang Buxin Dan,the platform incubation period and the total swimming distance of the rats were significantly better than those in the model group,but they still did not reach the normal rat level in the control group.Compared with the control group,the number of rats crossing the platform decreased significantly(P<0.01),while the target area movement time decreased significantly(P<0.01),and the first arrival time was significant extended(P<0.01);the number of times of crossing the original platform and the time of target area in each group of traditional Chinese medicine increased significantly compared with the model group(P<0.01),and the time of first crossing the platform was significantly shortened(P<0.01).During the water maze experiment,the Chinese medicine group rats can cross the platform individually,and the model group rats can rarely cross the platform.Experimental results of autonomous activity: the number of activity and the distance of activity in the model group were higher than those in the control group(P<0.01).In the low,middle and high dose group,the number of activity,the distance of activity and the time of activity were decreased(P<0.05).3.Immunofluorescence staining: the number of neurons,compared with the control group,the number of neurons in the model group was significantly reduced and the arrangement was not uniform;compared with the model group,the number of neurons in the Tianwangbuxindan group increased to a certain extent,and the high-dose group The increase was most pronounced,followed by the middle dose group.Compared with the control group,the IOD value of the model group was reduced.Compared with the model group,the Tianwangbuxindan low,medium,and high and low dose groups were increased to varying degrees,and the high dose group had the most significant increase.The middle dose group Secondly(P<0.05,P<0.01).Compared with Density control group,the model group decreased significantly compared with the control group.Compared with the model group,the low dose group decreased,the middle dose group did not change significantly,and the high dose group increased significantly.4.TUNEL staining: Compared with the control group,the marker index and apoptotic index of the model group were increased,indicating that sleep deprivation may cause apoptosis of hippocampal neurons;compared with the model group,the apoptosis index of the low and medium dose groups was relatively The decrease was more obvious in the low-dose group,while there was almost no change in the high-dose group,indicating that Tianwangbuxindan may improve neuronal apoptosis(P<0.01).5.RT-qPCR: can be seen from Table 7,compared with the control group,the m RNA transcription level of inflammatory factor IL-21 in hippocampus of rats in the model group was increased(P<0.01).after administration,the m RNA transcription level of each group decreased compared with the model group,in which the low and middle dose groups were still higher than the control group,and the high dose group was lower than the control group(P<0.01).the mRNA transcription level of hippocampal apoptosis regulatory factor Fas、Fas L、Caspase-8 was increased in the model group compared with the control group(P<0.01).after administration,the m RNA transcription level of apoptosis regulatory factor Fas、Fas L、Caspase-8 decreased in the chinese medicine group compared with the model group,among which the low and middle dose groups were still higher than the control group,and the high dose group was lower than the control group(P<0.01).The m RNA transcription level of regulatory factor Bcl-2 decreased significantly in the model group compared with the control group(P<0.01).All the high dose groups increased,among which the low and middle dose groups were still lower than the control group,and the high dose group was higher than the control group(P<0.01).6.Western Blot: The protein expression level of inflammatory factor IL-21 in hippocampus of rats: the protein expression level of IL-21 in the model group was significantly higher than that in the control group,and the protein expression level in the traditional Chinese medicine group was lower than that in the model group,and the higher the dose,the more obvious(P<0.01).protein expression levels Fas/Fasl apoptotic regulatory factors: compared with the control group,the expression level of the model group was significantly increased;compared with the model group,the protein expression level Fas/Fasl the traditional Chinese medicine group was decreased,and the high dose group was the most obvious,followed by the middle dose group(P<0.01).Adjustments The protein expression level of the control factor Caspase-8: compared with the control group,the expression level of the model group was significantly increased;compared with the model group,the protein expression level Caspase-8 the traditional Chinese medicine group was decreased,and the higher the dose,the more obvious(P<0.01).regulatory factors Bcl-2 protein expression levels: the expression level of Bcl-2 protein in the model group was significantly lower than that in the control group;the protein expression level Bcl-2the traditional chinese medicine group was higher than that in the model group,the high dose group was the most obvious,followed by the middle dose group,but the histone expression level of traditional chinese medicine was still lower than that in the control group(P<0.01).Conclusion:1.The heart for a living,is the main body of the five viscera,god dominates the function of life,is the leading basis of sleep function;sleep regulation,the mind played a vital role,and with this also Yingwei qi and blood and the function of the five viscera god normal operation.Therefore,the method of nourishing the mind and tranquilizing the mind is of great significance in regulating sleep.2.Intraperitoneal injection of PCPA insomnia rat model is reproducible.3.The selected Yangxin Anshen Fang Tianwang Buxindan has a sedative and hypnotic effect and can improve the sleep state of insomnia model rats.4.While improving the sleep quality of model rats,Yangxin Anshen Method can also improve learning and memory.Its mechanism may be related to the regulation of IL-21-mediated inflammation and Fas / Fas L-mediated neurons in insomnia in hippocampus Related to apoptosis. |
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