| Objective:Look up relevant document literature,understand the structure and function of the heart,the classification and function of mind and the relationship between heart and mind from the viewpoint of traditional Chinese medicine.Recognize the cognition of traditional Chinese medicine to insomnia from the etiology,pathogenesis,treatment,prescription and so on.Explore the relationship between mind and sleep,and the theoretical basis of treating insomnia by nourishing the mind and tranquilizing the mind.Establish rat model of insomnia and observe the effect of Yangxin Anshen method on insomnia rats and the effects of brain related nerve factors and their receptors.Through the theoretical and experimental research,this provides the theoretical basis and experimental basis for the application of the method of nourishing the mind and tranquilizing the mind in insomnia.Method:1.Theoretical discussion: Search information,Collate documents and know the systematic cognition between heart and mind from traditional Chinese medicine.And sum up the relationship between heart,mind and sleep.Make clear the mind's effect on sleep and explore the effect of nourishing the mind and tranquilizing the mind from traditional Chinese medicine.And finally sort out the relevant treatment principles and prescription.2.Experimental study: Fifty SD male SPF rats were selected to establish the insomnia model by multi platform water environment with intraperitoneal injection of caffeine and cytoxan,to observe the effect of modeling method of multi platform water environment with intraperitoneal injection of caffeine on rats' physique and related praxeology to observe the effect of modeling method of multi platform water environment with intraperitoneal injection of caffeine on rats' physique and related praxiology;Each group were took orally for 14 days,using Morris water maze to test the ability of learning and memory,using spontaneous activity test video analysis system of rats to observe autonomic activities;using TEM to observe the ultrastructural changes of hypothalamus,concretely on SCN morphology,organelles etc.;using immunohistochemistry to compare expression level of blank group,model group,western medicine group and Chinese medicine group rats hippocampus,hypothalamus neurotrophic factor;using ELISA method to detect the rat serum,hippocampus and hypothalamus nerve factor NGF,BDNF and VIP expression;using Real Time-PCR to detecte the effect of rats hypothalamus and hippocampus NGF,BDNF,VIP receptor expression.Results:1.General condition: Before modeling(T=0,T=7),the rats' condition in each group were active,and sleep were regular,hair were lustrous.With the model time expanding,the model group began to appear over excited,some rats' hair were dark and gloomy,the claw were white or pale yellow.At the end of modeling,the model rats' excitability were decreased and the condition were malaise.Most rats' eyes were glassy,and lips,claws and nails were pale,the hair were shaggy and react were tardy.While the blank group in the whole process of modeling were in good health,the activity,sleep,diet were normal.Weight:During modeling,The body weight of rats in each group increased,and the blank group increased significantly.Before modeling and after one week adaptive feeding(T=0,T=7),the model group in weight compared with the blank group have no difference(P > 0.05).After modeling(T > 7),the weight of the model group was significantly different from that of the blank group(P<0.01).Later stage of modeling(T=21,T=28),the body weight of the model group was significantly lower than that of the blank group compared with the blank group,(P<0.01)and the weight gain of the model group slowed down and tended stable.Hemoglobin concentration:there was no significant difference during the first week between model group and blank group in peripheral hemoglobin concentration(P>0.05),after modeling(T>7),model group compared with the blank group was reduced in hemoglobin concentration(P<0.01),Test on 21 d,28d show that TCM group and Western medicine group compared with the model group were increased in the peripheral hemoglobin concentration,but it was still lower than the blank group(P < 0.05).2.Morris water maze test results:The rats in the model group on the platform latency and swimming distance was significantly prolonged compared with blank group(P<0.01);after treatment,Tianwang buxin Dan high,low dose group and estazolam group was significantly shorter than the control group on the platform latency and swimming distance,there was statistically significant difference between groups(P<0.01).The model group compared with the control group,the rats' frequency of crossing platform were reduced(P<0.01),the target quadrant time were reduced(P<0.01),the first time to reach the platform were prolonged(P < 0.01);after treatment,the Tianwangbuxin Dan high,low dose group,estazolam group were increased in the number of cross platform than model group(P<0.01),the target quadrant time were prolonged(P<0.01),first time to reach the platform were shorten(P<0.01).In the whole process,the model group rarely passed through the platform,and someone in the medicine group can pass through the platform.Autonomous activity experiment result:Compared with the blank group,the number of activities and the distance of movement of the model group were significantly longer than those of the blank group(P <0.01).After treatment,compared with the model group,the number of activities,the distance of movement and the moving time of medicine group were decreased(P<0.01,P<0.05).3.Electron microscope observa:The neurons of SCN are mainly oval,and nucleus are irregular.The nucleoplasm is larger and cytoplasm is less.Mitochondrial crista is normal,to lipofuscin(Lipo)in blank group,the size is normal,the number is moderate,the cisternae is flattened and arrangement is neat in general.The number and volume of Lipo in SCN in the model group is increased and mitochondrial crista is partially fractured compared with control group,and the dyeing is deepened,the cystic complex of Golgi is disordered and has extensive swelling and vacuolization.After treatment,The volume of Lipo are increased a little,and the dyeing is deepened slightly,some cystic cells are disordered.Immunohistochemical results:BDNF immunoreactive products were mainly concentrated around the nuclear membrane and were brownish yellow or brown,with uniform staining and cyclic expression.The positive cells were widely distributed in the hippocampus,especially in CA3,CA1 and dentate gyrus(DG).The distribution of NGF was concentrated around the cytoplasm,the color was brown,and the neurons were arranged in good order,especially in CA3,CA1 and DG.There was statistically significant between groups in IOD comparison,compared with the control group,model group IOD is decreased;compared with the model group,each medicine group IOD is increased.4.Elisa test results:The results of NGF,BDNF and VIP in hypothalamus and hippocampus were consistent.Compared with the blank group,the contents of NGF,BDNF and VIP in the model group were significantly decreased(P < 0.01).After treatment,compared to the model group,the concentrations of NGF,BDNF,VIP in each medicine group were higher than the model group,but was still lower than that of the control group(P<0.01,P<0.05).Among all the tests,there was no significant difference in serum VIP concentration of low dose group compared with model group(P>0.05),and about serum and hypothalamus tests in estazolam group and Tianwangbuxin Dan high dose group,the difference was not statistically significant(P>0.05).5.Real Time-PCR detection results:Hypothalamic test: compared with the blank group,NGF,BDNF,VIP receptor mRNA expression in hypothalamus among other groups were significantly reduced;after treatment,the NGF,BDNF,VIP receptor mRNA expression in Tianwangbuxin Dan low and high dose group and positive drug group were increased in several degrees compared to the model group.Hippocampus test: In Tianwangbuxin Dan low and high dose group and positive drug group,NGF,BDNF,VIP receptor mRNA expression were decreased compared with control group;at the same time,compared with the model group,Tianwangbuxin Dan low and high dose group and positive drug group in NGF,BDNF,VIP receptor mRNA expression were increased in different degree.Conclusion:1.Heart is the root of life,and is dominated in the viscera.Mind is the master of life activities,and is the basis of sleep.The mind plays a key role in the regulation of sleep,and is also guaranteed by the normal function of visceral mind,ying qi and wei qi,Yin-Yang meridian etc.The method of nourishing the mind and calming the nerves is an important way to regulate sleep.2.The rat model of insomnia induced by intraperitoneal injection of caffeine combined with multi-platform water environment sleep deprivation model is reproducible.3.Yangxin Anshen representative Tianwang Buxin Dan has obvious sedative hypnotic effect,it can improve insomnia model rats' sleep condition,regulate the disorder of sleep rhythm.4.The method of tranquilizing mind has an obvious advantage in improving sleep quality of rats.At the same time,it also has obvious improvement effect on learning and memory level.The mechanism may be related to regulating the expression of NGF,BDNF and VIP in the brain. |
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