Experimental Study On The Alterations Of Aquaporins And The Intervention Of Kakonein After Focal Cerebral Ischemia In Rat

Cerebrovascular diseases is usual diseases.The prevalence rate ,death rate and disability factor is high.Accompany with the increasing of living standard ,the incidence rate of cerebrovascular diseases increased year by year.The cerebral infarction is the 70% of cerebrovascular diseases.In that, the 60% is the MCAO.The cerebral infarction induced the intracranial edama immediatly.It will reach peak on the third day,and recovers gradually on the seventh day.If we can reduce the edema ,we will treat the diseaes effectively.The transport of water is the basis of edema.The transport of water through the membrane have two way:1) travel by the simple diffusion.2)travel by the mediatation of AQP. Under the pathological conditions of the edema,water travel by the mediatation of AQP importantly. We first create the MCAO model on the rats. Then(1) detect the changes of mRNA of AQP1--AQP9 by RT-PCR. (2)detect the change of mRNA of AQP4 by hybridazation in situ.(3)detect the changes of protein of AQP4-AQP5 by western blot.(4)detect the change of protein of AQP4-AQP5 by immunohistochemistry. We perform the experiment in order to research the involvement of edema and provide the reason of new mechanism of edema. Furthermore,we use the kakonein to interfere and compare with the group of ischemic rat.we do it in order to research the new treatment and develop the clinial value of kakonein. Part I Summary of documents. (1)The research development of the AQP (2)The research development of the cranial edema. part â…¡Establishment of the MCAO model We establish the MCAO model and evaluate it by the neurological scale and pathological analysis (microscope,electron microscpe).The results are as follows: we do the 181 model.In that, there are 19 models which are not satisfied.In the others,we evalulate by the neurological scale and pathological analysis (1)The neurological seale:At 6h ,1d,3d of ischemia, the neurological scale increase with time and then the neurological scale decrese with time at 3d ,5d,7d,14d of ischemia .The scale of the grorp which use the kakonein is lower than the group of ischemia.(2)Pathological changes:The changes of microscope;The infarction is mainly locate the area of surface layer of coping cortex ,Insula ,lateral of piriformarea ,Nucleus amygdalae,putamen ,hypothalamls and the CA1,CA2 ,CA3of the Hippocampus. In the group of 6h .The tissue swell.There are a lot of triangle neurons which deeply stain in the surface layor of coping cortex.There are disseminated ensinophil neurons in the pennumbra,In the group of3d,we detect the widespread edema .There are a lot of necrotic neurons. In the group of 5d and 7d ,the edema decreases gradually .The glial and capillary hyperplasia greatly. In the group which use the kakonein,the area of infarction is more small than the ischemic group .The edema is lighter than the ischemic group. Electro microscope:In the 6h group,the neuron cell cytolisis,the mitochondrion swells. The nuclear chromatin gathers to the edge. Conclusion: we duplicate the MCAO model successful.The success rate approach to 90%. Through the neurological scale and pathological analysis,we think the model can be used to research the focal ischemia. Part â…¢: The alternation of the AQP after focal cerebral ischemia in rats . (1) detect the changes of mRNA of AQP1--AQP9 by RT-PCR. (2)detect the change of mRNA of AQP4 by hybridazation in situ.(3)detect the changes of protein of AQP4-AQP5 by western blot.(4)detect the change of protein of AQP4-AQP5 by immunohistochemistry. â… The mRNA of AQP1-AQP9 is detected by the RT-PCR:The results are as follows:There are no expression of AQP1,AQP2,AQP6,AQP7 in the brain. The expression of AQP4:In the control,there is the positive expression but the ambulance is weakly .In the 6h group,the expression increase obviously.It reach peak in the 3d group.Then,it decrease gradually .There is no obvious difference between the 14d group and the control group.The expressions of AQP5 and AQP9:The regularity of expression is as same as the AQP4, but the ambulance is more...