The Effect Of Consolidating Root Nourishing Brain To Expression Of C-FOS In Hippocampal In The Forgetful Model In Aged Rat

Objects:Establishing forgetful model to old age in rat by injecting D-galactose and nitrous acid sodium in the abdominal cavity.According to the tutor's experience who has treated the old and forgetful patients,to set up the three principles such as nourishing spleen and nourishing kidney and consolidating root, taking medicine to the model animals and to observe change of the memory,to clarify the mechanism that consolidating root nourishing brain may improve memory from the function and cytology and molecule biology,to define difference of the treating effects between nourishing spleen and nourishing kidney.To explain the science connotation of the theory on consolidating root nourishing brain and provide basis that Traditional Chinese Medicine(TCM)can improve memory and prevent brain functions from fading.To define the connection between the forgetfulness to old age and Spleen.Kidney,to explore epidemiology and pathogenesis on the forgetfulness to old age,to enrich and supply medical connotation on Spleen-Kidney in TCM.Methods:Through methods of studying documents,systematically considerate the theory about that ancient doctors cognized forgettery,look up the ancestor comprehension of forgettery,conclude physiology basis about memory and pathology mechanism about forgettery,according to Traditional Chinese medical theory,renewly summarize pathogeny and pathology mechanism on forgettery.Referring the experience on tutors treated forgetfulness to old age,according to pathogenesis on forgetfulness from all literature,filtrate medicine and compose prescription on nourishing spleen strengthening brain(Dangshen,Fuling,Shanzha,Shichangpu and so on),prescription on nourishing kidney and strengthening brains(Qiguo,Heshouwu,Shanzha,Shichangpu and so on), consolidating root nourishing brains(Dangshen,Qiguo,Heshouwu,Fuling,Shanzha,Shichangpu and so on),validate science and rationality of pathogenesis on forgetfulness through animal experiment.Previous research of the problem group had proved that the prescription could improve memory,but hadn't lucubrated the molecule mechanism and difference on treat effect of nourishing spleen and kidney.In order to define that how Spleen and Kidney impact memory,divide the prescription into the prescription on nourishing spleen strengthening brain and prescription on nourishing kidney strengthening brains,three prescriptions act on model animals,observe mechanism of treat effect,to clarify the mechanism three therapies may improve memory from the function and cytology and molecule biology.Filtering animals of memory with jumping,dividing into normal group with 15 and model group with 60.Injecting 2ml physiological saline into abdomen cavity of normal group every day,injecting D-galactose and into abdomen cavity of model group animals,successfully copy models after 40 days.Divide models into four parts,such as nourishing spleen group,nourishing kidney group,consolidating root group,model group,each of group is 15 averagely.The total is 5 groups including normal group,75 animals in all.Filling physiological saline 2ml every day with normal group and model group, filling liquor on nourishing spleen strengthening brain and nourishing kidney strengthening brains and consolidating root nourishing brains with other three groups.Dosage is 50g/kg/d or 40g/kg/d within 20 days.Beginning to behavior test after filling.Each of animal faste in 12h after test, draw blood from 1-10 number animals through decOllation,centrifuge 10min in speed of 3000r.min,adopting serum in term of specification in reagent-box.To menstruate SOD activity and MDA content.Adopt pallium and detect activity of Na~+-K~+-ATP enzyme.Adopt left-hippocampus and detect expression of C-FOS gene.Adopt right-hippocampus and detect expression of NR2A gene.Fasting 12h after behavior-testing with 11-15 number big rats of every group.Extract brains of rats and go along with immunity-coloration with biotin enzyme.Measure of immunity histochemistry ration is that hippocampus of rat brain is cut into 6 pieces,every piece is 10μm,adopt one from 3 pieces,separately use to colorate for antibody of NGF,NT-3,HSP70,namely adopt 2 pieces from each antibody of each big rat.Result:After making a model,the animal's fur is sparse,abscission,its eye gum increases,the amount of activity falls off,growth is slow,spirit is dejected,it is less than the normal group in drinking water and amounts of taking food,showing obvious old state,the characteristic is consistent with old animal.The Morrise's water maze test,the time that model group finds hiding platform is longer than normal group(P＜0.01).Time that three treating group finds the platform is shorter than model group,consolidating root group is the best(P＜0.05).Times that normal group and treating group go through platform location is more than model group after removing platform(P＜0.01 or P＜0.05).Consolidating root group is better than nourishing spleen(P＜0.01)and nourishing kidney(P＜0.05).Time that model group animal swims in platform location(C area)cuts down (P＜0.01)comparing with normal group.Time that treating group swims at C area is more than model group(P＜0.01 or P＜0.05).Consolidating root group is better than nourishing spleen and nourishing kidney(P＜0.01).Serum enzymes test displays,SOD of model group is lower than normal group (P＜0.01).Consolidating root group is higher than model(P＜0.05).Serum MDA of model group is higher than normal group.There are no differences among treating groups.Na~+-K~+-ATPase activity of model group in cerebral cortex is lower than the normal group obviously(P＜0.01);nourishing spleen group and consolidating root group is higher than model group(P＜0.01 or P＜0.05).NGF expression in normal group,nourishing spleen group and consolidating root group is higher than model group(P＜0.01 or P＜0.05).NGF expression in nourishing kidney is lower than consolidating root group(P＜0.05).NT-3 of the number of positive expression and gradation of positive cell is expressed is higher than model group(P＜0.05),is consistent with NGF result. Normal group has not positive cell of HSP70 immunoreaction in brain. Number of HSP70 immunoreaction cell of treating group is higher than model group(P＜0.01),consolidating root is the best.The number of HSP70 positive cell of consolidating root group is higher than nourishing spleen and nourishing kidney(P＜0.01).C-FOSmRNA expression of normal group is lower than model group(P＜0.01).Three treating group is higher than model group(p＜0.01),consolidating root group is higher than nourishing kidney obviously(P＜0.05),there is no significance with nourishing spleen(P＞0.05).There are certain amounts of NR2AmRNA expression in normal group,it is higher than model group(P＜0.05).NR2AmRNA expression of nourishing kidney group and consolidating root group is higher than model group(P＜0.01).Treating effect of consolidating root group which is higher than normal group is the best(P＜0.01).Conclusion:To analyze synthetically above result,problem group think that old age has a poor memory,which involves with multiple internal organs,it is a complicated illness related to qi blood sputum,but whose center is spleen and kidney.Making model of the forgetful old age animal by injecting D-galactose and nitrous acid sodium into abdominal cavity.Prescription of consolidating root nourishing brain can improve memory by various approach.For example,Rising to oxidation resistance,Rising energy supplies in cerebral cortex,strengthening neurotransmitter conduction,promoting renovation and regeneration of neuron,to resist neuron injury by disadvantageous factors and so on.consolidating root nourishing brain is important method treating forgetting old age.