Two Cisterna Magana Blood Injection Methods For Rabbit Cerebral Vasospasm Model And Comparative Observation

Objectives To establish a new simple, low mortality rate, high success ratesubarachnoid hemorrhage（SAH）model by the B-positioning percutaneous techniquesinfusing cisterna magna. To provide data support for exploring the most ideal animalmodels of cerebral vasospasm（CVS）after SAH by comparing the neurological score,the diameter changes and histological changes of basilar artery and basal vein atdifferent days after SAH of single and twice blood injection cisterna magna.Methods①A total of105New Zealand white rabbits were randomly divided intothree groups: control group (n=26), one-hemorrhage group (n=39),two-hemorrhagegrop(n=40), made by infusing the autologous arterial blood or normal saline into thecisterna magna at the guidance of type-B ultrasanic. CT scan is observed24h aftermodeling.②The animal behavior and neurological function were evaluated beforemaking model and1~11days after model made, the diameter changes of basilar arteryand basal vein were observed by magnetic resonance angiography （MRA） either.③The specimens of basilar artery and basal vein of each group were collected0~11days after SAH. Observing the wall morphological changes of basilar artery and basalvein by using HE stain and immunofluorescence stain.Results1. The method of infusing the cisterna magna at the guidance of type-Bultrasanic: no animals died in control group, one-hemorrhage group animal mortalityrate was5.13%, two-hemorrhage group animal mortality rate was7.5%, no significantdifference was found at each groups.24h CT shows the blood can be seen atsubarachnoid.2. The animal behavior and neurological function: Obvious symptoms ofneurological deficit were found0n3days after modeling at one-hemorrhage group andon5days after modeling at two-hemorrhage group; The symptoms of neurological deficit in two-hemorrhage group are more serious than one-hemorrhage group on5,7days.3. Rabbit cerebral vascular microsurgical anatomy and MRA contrast. Themicroanatomy shows that the basilar artery is walking in the middle of the ventralbrainstem just like the TOF-MRA image. The microanatomy of deep cerebral veinshows that no obvious boundaries between great cerebral vein and straight sinus wereseen just like susceptibility weighted imaging （SWI）image.4. The diameters of basilarartery and basal vein were observed by MRA between one and two-hemorrhage groupsafter modeling0~11days.①T he changes of basilar artery diameters: The diameters ofone-hemorrhage group become the most narrow on3days after modeling; While thediameters of two-hemorrhage group become the most narrow on5days after modeling.The diameters of basilar artery in two-hemorrhage group spasm more seriously thanone-hemorrhage group on5days and7days after modeling（P﹤0.05）.②The changesof basal vein diameter: The diameters of basal vein in both one and two-hemorrhagegroup become the most narrow on5days after modeling; The diameters of basal vein intwo-hemorrhage group spasm more seriously than one-hemorrhage group on3,5,7days after modeling（P﹤0.05）.5. The general histological observation of basilar arteryand basal vein.①HE stain of basilar artery: Obvious vasospasm changes of basilarartery wall such as endothelial folds and smooth muscle layer thickening were found on3,5days especially on3days at one-hemorrhage group; The more obvious vasospasmchanges of basilar artery wall were found on3,5,7days especially on5days attwo-hemorrhage group.②HE stain of basal vein: Basal vein wall is thin, only a singlelayer of cells surrounded the lumen, no wall sub-intima, intima, adventitia structurewere seen. No endothelial folds and wall thickening were found before and aftermodeling at one and two-hemorrhage group. The diameter is the most narrow in5daysafter the model.6. The immunofluorescence observation of basilar artery and basal vein.①vWF labeling of basilar artery: Obvious vasospasm changes of basilar arteryendothelial folds were found on3,5days especially on3days at one-hemorrhage group; The more obvious vasospasm changes of basilar artery endothelial folds were found on3,5,7days especially on5days at two-hemorrhage group. a-SMA labeling of basilarartery: The basilar artery wall a-SMA expression level increases on3,5,7days aftermodeling in one-hemorrhage group; more significantly increased appear on3,5,7daysin two-hemorrhage group.②vWF labeling of basal vein: vWF expression of the basalvein wall was negative before and after modeling at each groups. a-SMA labeling ofbasal vein: a-SMA expression of the basal vein wall was negative before and aftermodeling at each groups.Conclusions①T hemethod of injection blood to cisterna magna under B-positioningis a better way to establish the model of SAH.②The degree of vasospasm in the CVSmade by twice injection blood to cisterna magna is more intense and the time is morelasting than single injection blood to cisterna magna.③Basal vein can appearvasospasm after SAH on MRA image and histological like the basilar artery.