SAH大鼠抗脑血管细胞增殖研究

Experimental study of anti-cerebral vascular cell proliferation in Subarachnoid hemorrhage model rats

目的探讨大鼠SAH后脑血管细胞增殖变化,观察大鼠SAH后早期阻断VEGF的作用是否能减轻脑血管细胞增殖。方法 72只Sprague-Dawley大鼠随机分为3组,假手术组枕大池注入NS,模型组采用枕大池二次注血法制作SAH模型,干预组在制作SAH模型的同时通过枕大池注入抗VEGF多克隆抗体。于SAH(或NS)后1、3、5、7天时处死动物,分离脑干(包含基底动脉)做形态学和PCNA免疫组化检查,观测三组大鼠基底动脉的形态学改变、基底动脉壁PCNA表达。结果在SAH后第1天基底动脉即出现明显血管痉挛,第3、5天达到高峰,第7天逐渐缓解。SAH模型组大鼠基底动脉壁PCNA表达第1天即开始升高,第3、5天达到顶峰,第7天后逐渐下降;NS假手术组基底动脉壁PCNA呈阴性表达;抗VEGF抗体干预组基底动脉壁PCNA呈低水平表达。枕大池注射抗VEGF抗体能缓解SAH后脑血管肌内膜增殖,减轻脑血管壁的增厚程度。结论 VEGF介导的血管内膜增殖反应和脑血管壁增厚可能在脑血管痉挛的发生发展过程中起促进作用,SAH后早期应用VEGF拮抗剂能缓解脑血管细胞增殖。

Objective To explore cerebral vascular cell proliferation and observe the effect of VEGF to interrupt vascular cell proliferation in SAH model rats. Methods 72 Sprague-Dawley rats were randomly divided into three groups. Sham operation rats were injected normal saline into the cisterna magna. The SAH model rats were performed by use of the double hemorrhage model. The intervention group was injected anti-VEGF polyclonal antibody into cisterna magna before autologous arterial blood was injected into cisterna magna. Animals were sacrificed by euthanasia. The brain stem including basilar artery was obtained in order to perform hematoxylin and eosin stain and immunohistochemistry stain of PCNA. The mor- phological changes of basilar arteries of three groups were observed. The changes of expression of PCNA in the basilar artery wall were compared. Results The basilar arteries showed apparent vasospasm in SAH model rats at day 1 after the second injection of blood, and reached the peak at day 3 and 5, then gradually attenuated at day 7. The expression of PCNA in basilar artery wall in SAH model rats was increasing at the first day, peaked at day 3 and 5, gradually decreased at day 7. There was no expression of PCNA in sham operation group. The expression of PCNA in basilar atery wall in anti-VEGF polyelonal antibody intervention rats was obviously at lower levels. Injection of anti-VEGF polyclonal antibody into cisterna magna before injection of autologous arterial blood could block cerebral vascular cell proliferation in intima and could reduce cerebral vascular wall thicken- ing. Conclusion VEGF-mediated cerebral vascular cell proliferation in intima and cerebral vascular wall thickening may play a promoting effect in occurrence and progress of CVS after SAH. Treatment early with the VEGF antagonist can relieve cerebral vascular cell proliferation caused by SAH.