Lidocaine Treatment Of Delayed Cerebral Vasospasm And Neuronal Injury After SAH In Rats

Objective:To observe the cerebral vasospasm and changes in brain tissue, With SD rats of clean grade of secondary injection of blood into the cisterna magna model give lidocaine from the cistern in a different time to observe the cerebral, in order to understand lidocaine for subarachnoid hemorrhage and Late-onset cerebral vasospasm late after CA1 neural cells hippocampus whether area prevention and protection,about the prevention of delayed cerebral vasospasm after subarachnoid hemorrhage.Methods:1. Selected 60 clean grade of healthy male SD rats weighing 280 ~ 320g, no abnormal feeding after 1 week for the experiment.The experimental animals were randomly divided into four group: Control group, Sham group, SAH group, Lidocaine injection group.2. To Established animal model of subarachnoid hemorrhage, injecting autologous blood into cisterna magna in rats. About 30 minutes After the second injection of blood, Subarachnoid hemorrhage group and sham group were injected 0.1ml saline, and lidocaine was injected 0.1ml 2% lidocaine. Disinfecting incision suture, Then put mice put back in a rat trap,holding feeding. Free eating water. 3d, 5d, 7d after lidocaine injection, the model (mouse) perfusion, then take brian. And Placed in 4% paraformaldehyde solution saved.3. Various testing method:3.1 to observed changes about hippocampal CA1 area wide neurons and general structural change of the basilar artery by HE staining, then observe local cells from Micro-perspective.3.2 through the tar violet staining, Understanding neurons in hippocampal CA1 apoptotic changes.3.3 Using terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL) method. Determination of apoptosis of hippocampal CA1 neurons.3.4 using electron microscopy ,to observed nerve cells of the ultrastructural changes in various organelles about Hippocampal CA1 area.3.5 by immunohistochemistry, Determined Bax, bcl-2 apoptosis-related factors change for hippocampal CA1 area.3.6 By image analysis system to calculate the circumference of the basilar artery vessel diameter and vessel wall thickness, and observed changes in vascular structure.Results:1 The rat the hippocampus HE dyeing results :Normal controls and sham operation group to normal neurons form is visible. Neurons dyeing clear, neatly arranged rules, form complete, the nucleus is big and round, shading relatively shallow, located in cells, an occasional eosinophilic central nucleoli and cell bodies, axon, dendritic are visible, cytoplasm rich and evenly. Subarachnoid hemorrhage brain structure is very loose, the control group (present sieve, most change cells to break down, the nucleus of dissolved,and the second note blood first 3 days, after the first five days, the 7th day visible rupture cell and the nuclei of dissolved number increase gradually. Lidocaine injections group visible neurons nuclear broke into pieces, however,compared with the control group, subarachnoid hemorrhage, reducing the number of cells around the area of basic shape can be identified. And injection lidocaine for the first seven days after three days and 5 days than the decrease in the number of the abnormal cells is obvious.2 Tar violet stain: Normal controls and Sham operation group of big rat hippocampal nerve cell structure is clear, orderly rows, cytoplasm was stained lilac, nuclei is not shading, nucleoli shading. Subarachnoid hemorrhage of rat control of nerve cells in the hippocampus, the cells become pyknotic morphological deformation in cytoplasm decrescent, purple, nucleoli disappear, appear large necrotic cells. Lidocaine injections of nerve cells in the hippocampus structure disorder, cells reduce, a little cells form deformation, become pyknotic decrescent, cytoplasm hyperchromatic.3 TUNEL assay observations :Normal controls aligned nerve cells in the hippocampus visible light green uniform, the nucleus for fluorescence, the nucleus shape for the oval or round, individual cells appear nuclear marginalized and nuclear weeks local highlighted fluorescence phenomenon show cricoid.Sham operation group and normal controls no significant changes. Subarachnoid hemorrhage in control and lidocaine injections, group compared with the extension of time the apoptotic cells increases gradually, and lidocaine injections of group, the apoptotic cells relative reduction. Subarachnoid hemorrhage in control group (3d, 5d) and lidocaine injections group (3d, 5d) diam-eter phase group compared a statistically significant (P < O.05).4 Rats brain tissue transmission electron microscope observations :Normal control group and Sham operation group of nuclei and mitochondria saw no abnormalities. Subarachnoid hemorrhage in control and lidocaine injections of each phase of neural nuclei and mitochondria are notable changes have taken place, and bleeding is lidocaine injections control each phase of the nuclei and mitochondrial damage, especially with serious 7 day obvious.5 The rat the hippocampus Bax and Bcl - 2 expression :Normal controls and the Sham operation group bax expression for lower expression, BCL - 2 for high expression, and the two groups of comparisons was not statistically significant (P > O.05). But subarachnoid hemorrhage in control group (the first 3 days) started bax express enhancement, the first five days, the first seven days to express are high expression, positive cells increased and dyeing strengthened; Bcl-2 expression from three days to begin to express gradually decreased.Sham operation group with normal controls and compared subarachnoid hemorrhage phase in the group were a statistically significant (P < o. 01). Lidocaine injections group (the first 3 days) started Bcl - 2 express enhancement, 5 days, the first seven days to express are high expression, positive cells increased and dyeing strengthened; Bax expression from three days to begin to express gradually decreased. Sham operation group with normal controls and more pronounced in Billy phase injection groups are statistically significant group (P < o. 01) and control group (with subarachnoid hemorrhage 7d) and 5d, lidocaine injections group (5d diam-eter phase 7d), compared with the statistical significance (P < O.05), bax expression BCL - 2 decrease, express enhance, especially the first seven days, bax BCL - 2 to reduce significantly enhanced obviously.6 The rat basal artery HE dyed :Normal controls and sham operation group of basal artery lumen a round or oval, did not see the wall creases,lumen diameter of large area,long; Subarachnoid hemorrhage group of lumen is irregular, wall creases, lumen contraction obvious, the area is small, wall thickening, lumen diameter short; Normal controls and Sham operation group of the basal artery of inner perimeter and thickness of two groups of comparisons was not statistically significant (P > O.05). But subarachnoid hemorrhage from the first 3 days began to control diameter perimeter decrease gradually, wall thickness to increase gradually.Lidocaine injections from the first three days of inner perimeter gradually start decreasing, thickness of growth. Subarachnoid hemorrhage in control group (5d, with lidocaine 7d) injection group (5d diameter phase 7d), compared with the statistical significance (P < O.05).Conclusion:1,Lidocaine can alleviate after SAH late-onset cerebral vasospasm.2,Lidocaine can reduce rat hippocanlpal CA1 nerve cell injury and are obviously brain protection.3,Lidocaine can reduce CA1 area after SAH Bax nerve cell in the hippocampus gene expression,enhance the relative CA1 neural cells in the hippocampus Bcl-2 area gene expression ,reduce hippocampal CA1 neural cell apoptosis area.4,Because after subarachnoid hemorrhage nerve cells damage mechanism of not only confined to the late, and cerebral vasospasm may be some chemical substances stimulation, reason lidocaine there may be other protection mechanism, needs to be further study.5,Because this experiment to only within 7 days after SAH basal artery and nerve organizational change are studied ,whether for lidocaine for longer time inside of cerebral vasospasm, nerve cell injury and nerve function prevention and treatment effect,still need further research.