| Encephalopathy of ischemia is one of the diseases which injure people's health. Neurons died and relative functions are not sufficient durimg cerebral ischemia.In acute stage of ischemia, necrosis is the main way of neuronal death in the core of ischemic region,while neurons show secondary neuronal death around ischemic area. Apoptosis is obvious in this stage.Some genes' expression was associated with apoptosis.In resent,brain derived neurotrophic factors(BDNF) and Bcl-2,BAX played an important role in neuronal apoptosis. BDNF,Bcl-2 can keep neurons from damage,moreover, protect and recover neurons under ischemia.OBJECTS:Traditional Chinese Medicine(TCM) have certain curative effects in treating cerebral ischemia in recent years,but little is known about the mechanism.Di Huang Yin Zi is one of the Chinese herbs medicine applied in clinic.The purpose of this study was to research hippocampus' changes and expression of BDNF,Bax,Bcl-2 following ischemia in rats.We also investigated the effects of Di Huang Yin Zi in order to illustrate the mechanism of keeping neurons from injury.These results can supply the experimental bases for clinical application. METHODS:According to string method,build up a cerebral ischemic model of middle cerebral artery occlusion on Wistar rats.Each rat was anesthetized with 10% cholral hydrate(4ml/kgi.p).The animal was bound to an operation table.Cut skin in the middle of neck.Apart arteries on the right,including the common carotid artery(CCA), external carotid artery(ECA),internal carotid artery(ICA).Ligate the embranchment of ECA and CCA.Dissociate the trunk of ECA.Insert a fish line in it to creat the model of cerebral ischemia.By using H.E stain,observed the changes in hippocampus and the influence of Di Huang Yin Zi after ischemia. Useing method of inmmunohistochemic(ABC) to study BDNF,Bcl-2 and BAX expression of hippocampal neurons following cerebral ischemia .Steps as follows:First, Dewax paraffin slices, hydrate;Second,Slice was incubated in 1%H2O2-CH2OH under room temperature;Third,Slice was digest by protein enzyme K for 30 minutes in wet box under 37℃.4th,incubate 30 minutes in wet box under 37℃ with sheep serum.5th,Dripted antibody of BDNF(1:100),Bcl-2(1:100),Bax(1:100)on samples respectively and incubated for allnight in wet box.6th,take biotin of IgG and complex of ABC to incubate in wet box for 2 hours6. after that slice was taken in to solution of 0.05％DAB-0.01%molPBS for 30 minutes。7th,Visualize 5-20 minutes by DAB.8th,those steps need to be vibrated and bathed in 0.01MPBS solution except incubating by sheepserum.9th,dehydrate through alcohol whose concentration is up. make slice limpid by dimethylbenzene,envelop with gum.RESULT:1.In control group,vascular was dilatation around ischemic area.Neurons show denaturalization,such as cell body in alkalescence, nissle's body in dissolvimg, karyotheca crimpling,dark staining,hyperplasia glial cells and decreasing neurons.when the time of ischemia is extend,glial cells are greatly hyperplasic.At the same time,death of cells is more obvious and neurons become acidophil in part.In group of treatment with Di Huang Yin Zi,we can find that neuronal death is less serious than control group,neither is hyperplasia of glial cells.But neuronal death of treatment groups after nonage of ischemia is numerous, both of denaturalized cells and glial cells are increasing.2. There are evidences of positive neurons of BDNF in hippocampus of rats.The results show that BDNF-staining positive neurons changed with different time intervals after ischemia.The most number of positive cells was at 3d after ischemia. It is observed that the morphology of neurons was triangle or polygon.The nucleus were dark while neurons were losed. The live cells are decreasing when the time span of ischemia is extended in the control groups. On the contrary, changes can be improved in treatment groups, the morphology of neurons is similar to normal's. The live cells in treatment groups were more than in the control group.3.The res...
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