降钙素基因相关肽经鼻给药对蛛网膜下腔出血后大脑皮层细胞损伤的影响

Protection of Intranasal Delivery of Calcitonin Gene-related Peptide Against Cerebral Injury Following Subarachnoid Hemorrhage

目的探讨降钙素基因相关肽(CGRP)经鼻用药对蛛网膜下腔出血(SAH)后脑损伤的影响。方法将Wistar大鼠随机分为正常对照组、SAH组、经鼻生理盐水(NS)+SAH、经鼻CGRP+SAH组。用枕大池2次注入自体动脉血法制作SAH模型,CGRP和NS经鼻腔给予。于第2次枕大池注血后3 d,将大鼠处死取脑制作冰冻切片,用TUNEL法检测大脑皮层原位细胞凋亡,用免疫荧光结合激光共聚焦显微镜检测大脑皮层Bcl-2蛋白表达。结果解剖学观察表明,SAH模型制作成功。SAH组和经鼻NS+SAH组大脑皮层出现大量TUNEL阳性细胞,经鼻CGRP+SAH组大脑皮层TUNEL阳性细胞较经鼻NS+SAH组减少。SAH组和经鼻NS+SAH组大鼠大脑皮层Bcl-2蛋白表达明显增多,经鼻CGRP+SAH组大鼠大脑皮层Bcl-2蛋白表达较经鼻NS+SAH组增强。结论经鼻应用CGRP可减轻SAH后脑损伤。

OBJECTIVE To investigate the protective effects of intranasal delivery of calcitonin gene-related peptide（CGRP） on cerebral injury following subarachnoid hemorrhage（SAH）.METHODS Wistar rats were divided into normal control group,SAH group,intranasal NS＋SAH group and intranasal CGRP＋SAH group.SAH models were produced by double injection of autologous arterial blood into cisterna magna.CGRP and NS were given by intranasal perfusion.On the third day after second cisternal injection,the apoptosis of cells in cerebral cortex was determined by a TUNEL staining.The expression of Bcl-2 protein in cerebral cortex was observed by immunofluorescence method combined with laser confocal microscopic observation.RESULTS Anatomic observation revealed SAH models were successfully manufactured.A great deal of TUNEL positive cells appeared in cerebral cortex in SAH group and intranasal NS＋SAH group.The numbers of TUNEL positive cells in cerebral cortex in intranasal＋SAH CGRP＋SAH group decreased,compared with those in intranasal NS＋SAH group.Increased expressions of Bcl-2 protein in cerebral cortex in SAH and intranasal NS＋SAH group were found,compared with normal control group.The expression of Bcl-2 in intranasal CGRP group was more obvious than that in intranasal NS＋SAH group.CONCLUSION Intranasal delivery of CGRP may alleviate cerebral injury following SAH.