| IntroductionFor surgical operation sufferer to say,the occurrence which fears before the operation is a kind of widespread phenomenon.As a complicated stressor,fear emotion bring the physiological change meanwhile the psychological injure to the patient.How to repress the fear emotion effectively before the operation and make the patient could pass through the period surround operation safety is still need to resolve.Through the literature retrieval,the storage which knows terrified formation and memory all passes activate of quality with dopamine inside the system closely related. But the topic expect before the set of clinical trial result,ketamine compound the midazolam can the accurate decrease child fears before the surgical operation,and prove through an animal experiment its parts of function principleses include: influencing the expression of NMDA and GABA inside the cerebellum and hippocampus is be subjected to repressing the fear emotion and the storage of these memory.So whether applied ketamine compound midazolam can also influence dopamine system to represses the fear emotion.The conditioned reflex theory of the this experiment is based on the theories of classical,to establishment the fear rat model,and through observing the behavior result by using ketamine and midazolam,accepting the freezing%as the index sign which evaluates the degree of the fear rats,then carry on a histology slice to the rat brain, make use the immunohistochemistry skill to analyze the expression of the dopamine-1 receptor and the dopamine.Materials and Methods1.AnimalsMale,health,Wistar rats 112,are all provided by Chinese medicine university experimental animal department.2.Agent and EquipmentAgent:The keatamine and midazolam are provided by the ShengJing hospital anaesthesiological department;The rabbit anti-rat DA1R polyclone antibody and the SABC examination kit are bought from the BOSTER company;The mouse ant-rat monoclone antibody is bought from the Santa company and the SABC examination kit is from the Maixin company.Equipment:Self-made electric shock cage;Type XJ-2356 physiology stimulates generator;Type DIK-245 audio-generator;Microscope.3.MethodsThe behavior learns training:After measuring big rat a weight,they are equally divided into 4 groups,namely the normal sodium group(N group),katemine group(K group),midazolam(M group)and the compound group(K+M group).According to the classical conditioned reflex theory,we use a kind of low frequency tone to be the conditioned stimuli and the foot electrical shock to be a non- condition stimuli.Place rats in the electrical shock cage,after giving a 30 seconds of tone stimuli then shock their foot bottom 5 seconds.Repeat 4 times everyday,totally training for 5 days,and drug administration everyday conferrable correspond.Observe the behavior index on the 6th day to evaluate the degree of fear.The immunohistochemistry partment:At the day when we over the behavior training,we get the brain tissue at the 0 mins,10 mins,30 mins,60 mins,120 mins, 180 mins and 360 mins.Apply the step provided by the description to carry out the immunhischemistry experiment.4.AssessmentWe use the freezing%as the behavior index to evaluate the degree of fear,while freezing behavior is no body move in addition respiration and heart beating.Its covariance method is to take 10 seconds as one time of observation segment,if have any body movement we record "0",unless record "1" for the freezing,the percentage which finally appears freezing behavior inside segment by 10 times is finallyto observe index.The immunohistochemistry Assessment:When see the gray yellow grains appear as the positive result and random select 5 visual fields under the high power microscope(400×)then measure light density,take it as the immunohistochemistry index.5.Statistical analysisStatistical methods t-test were performed by SPSS software version 13.0.Results1.Freezing%The average freezing%of N group is 74%±16%,the K group is 49%±12%,the M group is 23%±10%,and the K+M group is 15%±10%.The freezing%of control group is significantly higher than the experiment group,and the P＜0.01.2.The expression of DA1R in the prefrontal cortex,midbrain and hippocampusThe expression of DA1R of the N group,K group,M group and K+M group in the prefrontal cortex,midbrain and hippocampus respectively is 0.5565±0.0215, 0.5121±0.0278,0.5362±0.0263;0.3877±0.0248,0.4245±0.0247,0.4195±0.0216; 0.3959±0.0235,0.4288±0.0300,0.4232±0.0252;0.3981±0.0260,0.3896±0.0203, 0.4031±0.0254,the P＜0.01. 3.The expression of DA in the prefrontal cortex,midbrain and hippocampusThe expression of DA of the N group,K group,M group and K+M group in the prefrontal cortex,midbrain and hippocampus respectively is 0.6272±0.0241, 0.5905±0.0259,0.5755±0.0235;0.4993±0.0229,0.5040±0.0235,0.5195±0.0224; 0.5000±0.0256,0.4868±0.0231,0.5047±0.0238;0.4162±0.0264,0.4754±0.0223, 0.4573±0.0219,the P＜0.01.4.The expression of DA1R and DA on each time pointFrom each time of expression of DA1R and DA,we observe the time when 30 minutes after drug administration is lower than the other time points.Conclusion1.The ketamine and midazolam could depress the fear behavior of rats,and eliminate the fear memory.2.The ketamine and midazolam could significantly depress the expression of DA1R and DA in the prefrontal cortex,midbrain and hippocampus.3.The effect of depression to expression of DA1R and DA is appeared at 30 minutes after taking orally.
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