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目的探讨蛛网膜下腔出血 (SAH)后血管内皮生长因子(VEGF)受体的变化。方法采用Wistar大鼠雌雄各半 ,将动物随机分入非SAH组和SAH组。非SAH组于股动脉抽血0.4ml后 ,枕大池注入0.3ml生理盐水 ;SAH组于股动脉抽血0.4ml后 ,枕大池注入经冻 -溶方法制备的自体动脉血溶解物0.3ml。每组各12只用于RT-PCR检测 (VEGF受体flt-1上游引物 :5’ -GACGGACTACCTGTCAATCA -3’ ;下游引物 :5’-GTCGCAAATCTTCACAACA -3’ ,目的片段长度748bp;β-actin游引物 :5’ -GGGAAATCGTGCGTGACAT-3’ ,下游引物 :5’ -CAGGAGGAGCAATGATCTT-3’ ,目的片段长度 :386bp) ,用计算机凝胶图象分析系统摄像并进行吸光度测定 ,得到VEGF或flt-1产物OD值与β-actin产物OD值的比值。各6只于脑池注射结束后72h行免疫组化检测VEGF受体flt-1和VEGF受体flk -1。结果RT-PCR结果显示非SAH组脑组织flt-1mRNA有很微弱的表达 ;SAH组在脑池注射后24hflt-1mRNA明显表达 ,72h表达强度进一步增高。非SAH组未见flt-1阳性细胞。SAH组和溶媒组在脑池注入动脉血溶解物后72h脑组织flt-1蛋白表达明显增强 ,其阳性细胞主要分布在大脑皮层 ,基底节区、海马等部位也有较多flt-1阳性细胞 ,脉络膜表达明显 ;flt-1蛋白主要表达在胶质细胞、微血管内皮细胞 ,部分神经元也见flt-1蛋
Objective To investigate the changes of vascular endothelial growth factor (VEGF) receptor after subarachnoid hemorrhage (SAH). Methods Wistar rats were randomly divided into two groups: non-SAH group and SAH group. In the non-SAH group, 0.3 ml saline was infused into the occipital cistern after drawn 0.4 ml femoral artery. In the SAH group, 0.3 ml autologous arterial lysate prepared by the freeze-thaw method was injected into the occipital cistern after blood drawn 0.4 ml in the femoral artery. 12 of each group was used for RT-PCR detection (VEGF receptor flt-1 upstream primer: 5 ’-GACGGACTACCTGTCAATCA -3’; downstream primer: 5’-GTCGCAAATCTTCACAACA -3 ’, target fragment length of 748bp; β-actin primer : 5’-GGGAAATCGTGCGTGACAT-3 ’, and the downstream primer: 5’-CAGGAGGAGCAATGATCTT-3’, the target fragment length: 386 bp). The image was taken by a computer gel imaging system and the absorbance was measured to obtain the OD value of the VEGF or flt-1 product And β-actin product OD value ratio. The expression of VEGF receptor flt-1 and VEGF receptor flk-1 were detected by immunohistochemistry in each 6 rats 72h after the end of cistern injection. Results The results of RT-PCR showed that there was a weak expression of flt-1 mRNA in brain tissue in non-SAH group. The expression of 24hflt-1 mRNA in brain tissue of SAH group was significantly increased at 72h after injection. Non-SAH group no flt-1 positive cells. The expression of flt-1 protein in brain tissue of SAH group and vehicle group was significantly increased at 72h after injection of arterial lysate into the cerebral cistern. The positive cells were mainly distributed in the cerebral cortex, basal ganglia, hippocampus and other sites also had more flt-1 positive cells, Choroidal obvious expression; flt-1 protein is mainly expressed in glial cells, microvascular endothelial cells, some neurons also see flt-1 egg