blood; there were CGRP and ETRA immunohistochemical positive staining granula expressions in the basilar artery tissues. Its dynamic change regularity was as follows: the expression of CGRP was the most obvious at 1 h, and decreased from day 3 to day 5 by turns, then on days 5 reached the lowest; it began to increase from day 7, the expression on day 10 was relatively obvious, but did not reach the normal level. There was scattered irregular positive staining in the normal saline group and puncture group. ETRA was expressed on the basilar artery both in blank group and puncture group. They were expressed mainly on the vascular endothelial cells. ETRA was also expressed on the basilar artery in the saline group. ETRA was expressed heavily on the basilar artery’s endothelial cells in SAH group, mainly in 3-day subgroup. Smooth muscle cells also expressed ETRA. Conclusion The expression of CGRP and ETRA in the cerebral vessel cells after subarachnoid hemorrhage presents obviously dynamic changes; and it is suggested that CGRP plays a very important role in the delayed cerebral vasospasm.
KEY WORDS: subarachnoid hemorrhage; cerebral vasospasm; calcitonin gene-related peptide; endothelin receptor A; immunohistochemistry
迟发性脑血管痉挛(delayed cerebral vasospasm, DCVS)是蛛网膜下腔出血(subarachnoid hemorrhage, SAH)最常见的病理改变之一。但是，其发病机制目前仍不十分清楚。近年来的研究发现[1-3]，内皮素-1(endothelin-1, ET-1)可以特异性的和内皮素受体A(endothelin receptor A, ETRA)结合，引起血管的强烈收缩，而降钙素基因相关肽(calcitonin gene-related peptide, CGRP)却具有拮抗ET-1的收缩作用。但是，目前有关CGRP的相关研究均测定的是血液或脑脊液中CGRP水平的变化[4-5]，而其在脑血管内皮上的表达情况，尤其是与ETRA表达的对照研究较少。本实验采用枕大池二次注血法建立兔SAH后DCVS模型，旨在探讨SAH后CGRP与ETRA在在脑血管表达的动态变化规律，进一步阐明SAH后DCVS的作用机制。
1 材料与方法
1.1 材料
实验动物为日本大耳白家兔，60只，雌雄不拘，健康状况良好，体重2.1-2.9(2.5±0.4)kg，在相同的条件及环境中(饲料和温度均相同)饲养一周后进行实验。所有实验用兔均购自西安交通大学医学院实验动物中心。
实验试剂及药品：CGRP一抗、兔抗兔ETRA单克隆抗体、SABC试剂盒、DAB显色试剂盒均购自武汉博士德生物工程公司;40 g/L多聚甲醛灌注液购自西安交通大学医学院器材科;250 g/L乌拉坦溶液、0.01 mol/L枸橼酸盐缓冲液、PBS缓冲液以及修复液均由西安交通大学医学院组织胚胎与解剖实验室提供;手术器械等由西安交通大学医学院第
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