| Background and ObjectiveAs a scaffold protein, Axin has many domains which can directly or indirectly interact with corresponding proteins and plays critical role in embryonic development,tumorigenesis and cell apoptosis.Axin involved in regulating Wnt,JNK,p53 and other signaling pathways ,and overespression of Axin can promote cell apoptosis.Expression and significance of Axin has been reported in many malignant tumors and culture cells,but its expression rarely seen in the rat with middle cerebral artery ischemia reperfusion injury model.Our study used the model of middle cerebral artery occlusion in rats and examined the expression and significance of Axin through immunohistochemistry staining. Materials and Methods40 SD rats were selected and randomly divided into two groups:(1)sham operation(Sham group n=20);(2)ischemia reperfusion group (IR group n=20).According to reperfusion time ,the two groups were divied into 6h,12h,24h,48h four groups,each group had five rats.Surgery group produced by suture method model of ischemia-reperfusion injury,sham operation group with the same set of actions,but not to insert lines. After 2h ischemia ,both groups were reperfused at 6h,12h,24h,48h.Then the brains of each group were removed after the 4% paraformaldehyde perfusion.The parts that before and after the optic chiasm for 1mm were processed coronal section,and been fixed in 4% paraformaledhyde for 24h.After been washing in the running water for 12h,the parts were dehydrated,transparened t,embedded in paraffin blocks ,and sectioned into slices of 4μm.Subsequently HE staining, immunohistochemical staining were performed .Finally the expression of Axin located in hippocampal CA1 region and parietal cortex region were detected by using image analysis system . ResultsOur data shows : (1)neurological score:Sham group did’t found neurological deficits.Neurological deficits were observed in IR group in which 24h subgroup was the most obvious.(2)the changes of HE staining brain tissue .Sham group :the nerve cells which located in hippocampal CA1 region and parietal cortex region have normal morphology,integrate structure ,closely array,and clearly border .we also found that the neurons have lightly stained cytoplasm ,blue nucleus which shaped round or oval and has clearly outline.IR group:the neuron of ischemic hippocampal CA1 region and parietal cortex region has disordered arrange,concentrated plasma which even formed cytoplasmic vacuoles,concentrated nucleus .We also found that interstitial were thin and the number of normal nerve cells were reduced.(3)The sections of Axin which processed immunohistochemical staining were detected in the light microscope.Sham group:The expression of Axin in hippocampal CA1 region and parietal cortex region can be observed .We found that the positive performance of neuron located in the cytoplasm.IR group: The expression of Axin which located in hippocampal CA1 region and parietal cortex region was more obviously than that of Sham group. The two groups have significantly different(P<0.05). IR group :the expression of Axin at different time of reperfusion have significant difference (P<0.05) ,in which the expression of 24h subgroup higher than other subgroups.Conclusions⑴Firstly revealed the expression of Axin in rat with middle cerebral artery ischemia reperfusion model by immunohistochemistry.⑵The expression fo Axin increased after cerebral ischemia reperfusion injury,and the expression is consistent with the apoptosis situation of the model,which suggest that Axin may be involved in apoptosis mechanism of cerebral ischemia reperfusion injuryand maybe through the mitochondrial pathway related death.⑶The positive expression of Axin was located in the cytoplasm of rat brain。... |
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