Proteomic Analysis Of Synaptosomes Of Subarachnoid Hemorrhage Rats With Cerebral Vasospasm

Objective1. To research the dynamic state of synapsinⅠand p- synapsinⅠin SAH rat brain,and to investigate the effect of synapsinⅠand p- synapsinⅠin the process of SAH's generate and development.2. To construct the two-dimensional gel electrophoretogram of the synapsins proteome of the SAH rat brain.It could be the basis to find the disease-specific proteins and to research the mechanism of the CVS after SAH.Methods1. In this study, Methods Healthy Wistar rats were used in the experiment. Autologus arterial hemolysates were injected into rat cisterna magna to induce SAH. The animals were randomly divided into sham operated and SAH groups,and each group was randomly divided into 0d,1d,3d,5d and 14d groups.To observe the diameter of the basilar artery(BA) by Somatotype microscope and livingbody circulation measure system.The distribution and the dynamic expressions of synapsinⅠand p- synapsinⅠwere observd by optical microscope after immunohistochemistry staining.2. In this study, Methods Healthy Wistar rats were used in the experiment. Autologus arterial hemolysates were injected into rat cisterna magna to induce SAH. The animals were randomly divided into normal control,sham operated,SAH +CVS1h,1d,3d,5d,14d groups.The synaptosomes of cortex and hippocampus were extracted by density gradient centrifugation ,then to analyze the expression of synapsinⅠand p- synapsinⅠamong the groups by Western blot. 3. In this study, Methods Healthy Wistar rats were used in the experiment. Autologus arterial hemolysates were injected into rat cisterna magna to induce SAH. The animals were randomly divided into normal control and SAH+CVS 3d groups. The synaptosomes of cortex and hippocampus were extracted by density gradient centrifugation ,then clearaged and purificated.Two-dimensional electrophoresis was performed on serum protein of both groups.Result1. Compared with sham operated group,solvate widespread in subarachnoid space and obviously spasm of basilar artery could be seen in SAH rats'brains.Deposits of synapsinⅠwere mainly distributed in neuropile,the expression of synapsinⅠin cerebral cortex was little,which in the regions of CA1-CA4 of hippocampus was more,and especially in the regions of hilus fascia dentata and CA4 was the most. SynapsinⅠ's expression of SAH rats was lower than sham operated rats at all the time point, the decrease is not clear at the 1 hour after operation, and obviously decrease was observd at the 1d time point,the expression was the lowest at 3 day.(P<0.01),and then recovered gradully,to achieve the level of sham operated group at 14 day.The dynamic expressions of p- synapsinⅠwere similar with those of synapsinⅠ,but the expression was lower than synapsinⅠ,and the decrease was more significant .2. To extract the synaptosomes of cortex and hippocampus by density gradient centrifugation could successfully build the 2-DE images of the synapsins proteome of SAH rats.The result displayed that the PI of the synapsins was mainly between 4 and 7. After contrastive analysis, we found out 13 disparity proteins between the samples of cerebral cortex,and 12 disparity proteins between the samples of hippocampus.Conclusion1. Autologus arterial hemolysates injected into rat cisterna magna could induce stably modle of SAH ,and obvious cerebral vessels spasm could be observed.2. The expressions of synapsinⅠand p- synapsinⅠof cerebral cortex and hippocampus all decrease obviously after SAH,and the decrease at 3 day after SAH is the most significant.The results illustrate that synapsinⅠand p- synapsinⅠare related with the CVS after SAH. 3. The synapsins proteome's 2-DE images of SAH rats were successfully built,and it is very useful for searching specificity proteins associated with SAH.