| Objectives1.To investigate the effect of rhG-CSF given intranasally on cerebral vasospasm following SAH.2.To investigate the effect of rhG-CSF given intranasally on relieving acute cerebral vasospasm of rats following SAH.3.To investigate the effect of rhG-CSF given intranasally on brain injury secondary after subarachnoid hemorrhage.Methods:1.Twenty-four adult healthy male Wistar rats were divided randomly into four equal groups(n=6), including normal control treated group,SAH group(treated with hemolysate), NS+SAH group(normal saline given intranasal-lly before operation), rhG-CSF+SAH group(hemolysate given intranasally before operation).Animals were treated with either artificial CSF or hemolysate. Left craniotomy was performed and BA was posed. Somatotype microscope and livingbody circulation measure system were used to observe the caliber of BA. Pial vasculature were posed and observed by olymPus biological microscope. Pial microarterial and microvenous diameter changes were recorded by video record.2.SAH model was produced by double intracisternal injection of freshly autologous arterial blood. Twenty-four adult healthy male Wistar rats were divided randomly into four equal groups(n=6), including normal control treated group SAH group,NS+SAH group, rhG-CSF+SAH group.All animals were sacrificed at 72h after SAH by perfusion.The brain along with basilar arteries(BA)were collected and made into the paraffin sections.By HE staining to detect morphological changes of BA; Immunohistochemical analysis on expressions of Caspase-3,NF-kb in Rat's cerebral cortex and hippocampus; Nissl's staining and image analysis were taken to estimate number of dead neural cells, optical density of positive cell of Nissl's staining of hippocampus CA1 area and cerebral cortex in rat brain.Results1.The BA in SAH group and NS+SAH group had obviously cerebral vasospasm,the caliber was significantly coarctat compared with that in normal control treated group(p<0.01); the BA in rhG-CSF+SAH group slightely cramped,the caliber of BA was lightly coarctat compared with SAH group and NS+SAH group(p<0.01). Pial vasculature in SAH group and NS+SAH group were obviously abnormal,blood flow much presented flow of sediment,even could be found blood stasis and movement2.(1)By HE staining, the basilar artery revealed luminal narrowing, reduction in diameter,thickness of wall, corrugation of the internal elastic lamina (IEL), and cellular proliferation were observed in the spastic BA of the SAH group and NS+SAH group; the BA in rhG-CSF+SAH group was reliever than the SAH group and NS+SAH group. (2)By immunohistochemical analysis, Caspase-3 and NF-κB positive reaction increased obviously in cerebral cortex and hippocampus of the SAH group and NS+SAH group. In rhG-CSF+SAH group the expresstion of Caspase-3 and NF-κB were less than SAH group.(3) Nissl staining suggested in normal control treated group the color of Nissl staining was deeper,the number was more and the structure was more regular than that of the SAH group and NS+SAH group.In the SAH group and NS+SAH group, the staining was lighter and the structure was less regular than that of the control group, some pyramidal cells shrunk, their nuclear dwindled and presented triangles, and there were less or no Nissl's bodie.(neuronal cells were characterized by considerable swelling, and the empty vacuolar structures appeared in the cytoplasm).Neurons in CA1 there were more Nissl's bodies in the rhG-CSF+SAH groupConclusionsl.rhG-CSF via intranasal pathway could relieve basilar artery vasospasm;improve cerebral microcirculation;raise brain blood provision following SAH. 2. rhG-CSF given intranasally can significantly reduce the expresstion of Caspase-3,a proapoptotic factor, than the SAH group; can suppress the activation of NF-κB effectively and reduce the death of neural cells; can decrease nerve cell death and apoptosis of hippocampus area to ameliorate brain injury secondary after subarachnoid hemorrhage. |
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