| Cerebral vasospsam(CVS) is a major cause of death and disablement in pateits with aneurysmal subarachnoid hemorrhage (SAH). Past research on CVS concentrated to the platelet Products (5-Serotonin, angiotensin, catecholamine and K+) , erythrocyte decomposition Products (Adenosine triphosphate and oxyhemoglobin), unbalance between Nitrogen monoxide and endothelins and disorder of ionic channel, but it is unclear that molecule mechanism of persistent contraction of smooth muscle and inflammatory reaction in CVS, especially, the rule and relation of the expression and activity of PKC and NF-κB. In our study, We used two-hemorrhage model by injection of autologous aterial blood into cisterna magna on 0d and 3d to initiate the pathological and molecular changes of basilar artery associated with SAH, and evaluated the expression and activity changes of PKC, NF-κB and IκB by immunohistochemical,substrate phosphorolysis kinase assay, Western blot analyses and electrophoretic mobility shift assays (EMSA). Meantime, We apply Myristoylated PKC Peptide inhibitor to prevent and cure CVS in two-hemorrhage model in rabbits, and discuss the molecular biological mechanism that Myristoylated PKC Peptide inhibitor effect on CVS, and offer a theoretical basis to prevention and cure CVS . Our study were devided into five section as follow: 1 Experimental study of morphologic changes of basilar artery in two hemorrhage model in rabbitsThe objective of this study was to observe morphologic changes of basilar artery in two hemorrhage model in rabbits,and offer the data of morphologic changes for studying mechanism and intervention result on CVS . 35 male chinese white rabbits, weighing between 1.5~2.0kg, were used in this study and randomly divided into two groups: ①control group(n=5);②operation group(n=30):two-hemorrage model of cerebral vasospasm was constructed by injection of autologous arterial blood into the cisterna magna on 0d and 3d. Five animals of operation group were killed by perfuse 4% paraformldehyde buffer solution into aorta (90mmHg) on 1d, 3d, 5d, 7d, 10d and 14d, rapidity take out basilar artery including brain stem and cut upper half of basilar artery, and fixed for 24h in 4% paraformldehyde, paraffin embedding and serial section of 5μm, carry on hematoxin eosin(HE) and masson's trichrom stain to observe the morphologic changes by light microscopy and measure inner perimeter and artery wall thickness; another half of basilar artery were fixed 24h in 2.5% glutaraldehyde, epoxy resin embedding, ultra section to observe ultra- structure by transmission electron microscopy(TEM).Light microscopy: The structure of basilar artery is no abnormality including thin collagenous fiber in adventitia and a little extracellar matrix in smooth muscle in control group 1d and 3d after injection of blood. We observed slightly arterial narrowing, corrugation of endothelial cells, hypertrophia of smooth muscle cells, extracellar matrix in smooth muscle layer begin to increasing and collagenous fiber in adventitia become thicking and monocyte and neutrophile granulocyte infiltration in adventitia in basilar artery on 5d. On 7d after injecting blood, severe arterial narrowing,corrugation and disruption of endothelial cells, hypertrophia, distortion and necrosis of smooth muscle cells, more extracellar matrix in smooth muscle layer, collagenous fiber and monocyte and neutrophile granulocyte infiltration in thick adventitia were found. 10d after injection of blood, arterial narrowing, hypertrophia of smooth muscle cells and proliferation of extracellar matrixbegin to relieve, and remained disruption of endothelial cells, proliferation of collagenous fiber inflammation reaction weaken in adventitia. To 14d ,there are only light corrugation of endothelial cells and a little necrosis of smooth muscle cells.TEM: The structure of basilar artery is no abnormality in control group. On 1d and 3d after injecting blood, light vacuolar degeneration of endothelial cells were observed. On 5d, it were found that...
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