Study On Ethological Effects And Its Mechanism Of Acupuncture On Hypoxic-ischemic Brain Damage In The Neonatal SD Rats

PartⅠThrough comprehensive researches on quantity of documents on Hypoxic-ischemic Brain Damage(HIBD),partⅠcan be divided into two parts as fellow:The first part,an intimate analysis has been made on the current research status of how to identify the definition,symptoms,etiopathogenesis and treatments,etc.of Hypoxic-ischemic Brain Damage in TCM,and especially the development of acupuncture treatment for HIBD throughout the world.It is considered that acupuncture is one of the effective therapies for treating HIBD,which builds a substantial clinical foundation for our empirical study.The second part,an overview has been made on the research for HIBD of modern medicine.The mechanism of HIBD is very complicated.For the pass few years,the research on its mechanism has been increasingly profound,though the exact mechanism hasn't been wholly clarified so far.However,with the development of molecular biology,researches based on molecular level can be continued on cerebral anoxia and ischemia of neonats and neonatal animals, therefore has come new hope for the prevention and cure for HIBD.So far,there is no specific remedy for children' s brain diseases.Experts throughout the world have done lots of research,and have made some achievements.Among them, acupuncture effects have increasingly attracted international recognition, which worth further lucubration.PartⅡObjective:Test from behavior locomotory,learning and retention ability, apoptosis cells,apoptosis related gene protein Bcl-2,Bax expression,and HE pathomorphology,etc,in order to approach the possible mechanism of learning and retention handicap,ethologic handicap of the neonatal rats which are Hypoxic-ischemic Brain Damaged,and to approach the possible mechanism of the amelioration of the mentioned handicaps with the methods of acupuncture.Methods:Demesh and deligate both arterias uterina of the pregnant rats, wholly block the blood supply for 25min,then birth fetus rats,so the models of Hypoxic-ischemic Brain Damage are made.The successful-made models are randomly divided in to the acupuncture group and the model group,8 rats each. In normal group,during the surgery the pregnant rats' cervixes are clamped by haemostats,the fetus rats are taken out immediately,8 rats.After the acupuncture group has been treated therapy,ethology of the rats in each group are tested;then observe the pathomorphology changes of the animals with HE colored,and the changes in brain tissue cells apoptosis and apoptosis related gene Bcl-2,Bax expression.Results:1.The Ethology index1.1 Learning and retention abilities(Maze Y)Comparison of three groups of the number of error reactions(EN)and their differences revealed significant difference(ANOVA,F test = 17.45,P＜0.0001)before acupuncture(14d).And multiple comparison show significant difference between the normal group,the model group and the acupuncture group (P＜0.05),except the model group and the acupuncture group.The difference of interclass has statistical significance after further treatment(ANOVA, F test = 75.83,P＜0.0001).And Multiple Comparison again,there is significant difference found as compared with any two groups and there is statistical difference between the two groups.1.2 Grasping experimentsThe average time of rats grasping one horizontal plastic rope by the lead leg and the average score of the normal group,the model group and the acupuncture group are 30.19±15.03 and 2.5,12.19±6.37 and 1.75, 10.22±7.03 and 1.25 one by one.The difference of interclass has statistical significance(ANOVA).Then Multiple Comparison,the significant difference is found between the model group,the acupuncture group and the normal group (P＜0.05).There is no significant difference between the acupuncture group and the model group(P＞0.05).After acupuncture(33d),the average time of rats grasping one horizontal plastic rope by the lead leg in the normal group last 375.16±42.50 and the average score is 5;the model group is only 56.08±49.97 and 2.5;and the acupuncture group is 177.86±93.22 and 3.75.The differences are statistical significance in each two groups(both P＜0.05).1.3 Slope experimentsThe average expensing timeof the normal group,the acupuncture group and the model group is 7.13s,11.80s,15.15s one by one at the 14^thd.The differences were statistical significance between the model group,the acupuncture group and the normal group except the acupuncture group and the model group.At the 33^thd,the average expensing time of the normal group, the acupuncture group and the model group is 2.94s,5.78s,10.40s one by one. Significant differences in two groups are observed between the normal group, the acupuncture group and the model group(P＜0.05).However,there is no statistical significance compared with the normal group and the acupuncture group(P＞0.05).1.4 Playground experimentsThe average score before acupuncture(14d)of the normal group is 1.38, and There are no differences between the acupuncture group and the model group, both 0.75.After acupuncture(33d),the average score of the normal group, the model group and the acupuncture group is 1.88,1.38,1.75 one by one.Then compare the average score among the three groups both before and after acupuncture,the difference is not statistically significant.2.The effect of acupuncture on apoptosis of rat brain cellsThe apoptosis index of rat brain cells in the normal group rarely appear, only(1.0±0.21),but the model group is rather high(20.00±3.18).The difference was significant.The values of the acupuncture group(9.05±2.07) are among the normal group and the model group.Separately compared with the normal group and the model group,it is significantly different,respectively (t test,P＜0.05).3.The effect of acupuncture on expression of apoptosis-related Bcl-2 protein and Bax proteinIn the normal group,its bcl-2 protein express scarcely(0.51±0.24) and the quantity of bax protein is very low(1.03±0.27).But the expression of bax protein in the model group is significantly increased(1.54±0.17) and its bcl-2 protein is decreased(0.26±0.06).The model group compared with the normal group in bcl-2 protein,and the difference is significant(P＜0.05).The result is the same as between the acupuncture group and the model group(P＜0.01).In the bax protein,there are significant differences between the model group and the normal group(P＜0.05),the acupuncture group and the model group(P＜0.05).4.The effect of acupuncture on HE pathomorphologyThe nerve cells of cerebral cortex and hippocampus in the model group lost and degenerate the most seriously in the 3 groups.Although there are still varying degrees of nerve cell degeneration in the acupuncture group, but the number of the degenerated nerve cell is significantly reduced,some sporadic visible damage cells.The shape of the pyramidal cells of the hippocampus,is basically normal.Conclusion:1.Acupuncture can improve the learning and retention abilities(maze Y),balance ability(grasping experiment),reaction ability (slope experiment),of SD rats which is Hypoxic-ischemic Brain Damage.2.Minor intrauterine hypoxia and ischemia would cause apoptosis of neuronal cell in brain tissues indeed.And acupuncture treatment can restrain nerve cell from apoptosis which is caused by intrauterine hypoxia and ischemia.3.The stimulation of intrauterine ischemia can cause the expression of apoptosis-associated genes Bax-protein in neonatal mouse,and acupuncture treatment can increase the its Bcl-2 protein and decrease the Bax-protein, thus restraining nerve cell from apoptosis which is caused by intrauterine hypoxia and ischemia.4.The dysfunction or premature in the model group may due to the apoptosis of the nerve cells as insufficient blood to the brain within the uterus.The metal ability of the acupuncture group is better than the model group,which may due to the stimulation of acupuncture therapy by the protection of the nerve cells.