神经肽Y对蛛网膜下腔出血后脑血管平滑肌细胞增殖及α-actin表达的影响  

Effects of neuropeptide Y on the cerebral vascular smooth muscle cell proliferation andα-actin expression after subarachnoid hemorrhage

作　　者：薛俊刚[1,2] 宋锦宁[1] 安吉洋[1] 王军锋[1] 李丹东[1] 张明[1] 孙鹏[1] 李宇[1] 

机构地区：[1]西安交通大学医学院第一附属医院神经外科,陕西西安710061 [2]西安市红会医院神经外科,陕西西安710000

出　　处：《西安交通大学学报（医学版）》2013年第6期744-748,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家高技术研究发展计划"863计划"基金资助项目(No.2006AA02Z4Z4);教育部高等学校博士学科点专项科研基金(No.20110201110060);教育部新世纪优秀人才支持计划资助项目(No.NCET-05-0831)~~

摘　　要：目的观察神经肽Y(NPY)在蛛网膜下腔出血(SAH)后脑血管平滑肌细胞增殖及收缩蛋白表达变化,揭示NPY在SAH后迟发性脑血管痉挛中的作用。方法 SD大鼠150g,雌雄不拘,失血法处死并行生理盐水灌注,取大鼠Willis环动脉及其分支,行原代细胞培养并传代,向细胞培养基中加入自身全血使氧合血红蛋白终浓度达10-6mol/L以建立SAH血管痉挛平滑肌细胞模型并进行分组,包括空白组、SAH组、NPY组、NPY+SAH组、NPY+SAH+BIBO3304组(干预组)、NPY+BIBO3304组,培养1~5d后用免疫荧光法检测α-actin表达,IPP软件测荧光强度半定量,SPSS 16.0统计软件进行数据分析。结果应用消化法成功培养出血管平滑肌细胞(VSMCs),经细胞活度检查示活度>97%,经细胞纯化后细胞免疫染色未见其他种类细胞,经传代培养至第5代时可见细胞呈典型"谷-峰"样生长。细胞干预3d后NPY组与NPY+SAH组较SAH组与空白组α-actin荧光强度明显增强(P<0.05),而干预组与NPY+BIBO3304组α-actin荧光强度与空白组相比差异无统计学意义(P>0.05)。结论 NPY上调VSMCs的α-actin表达,而其受体拮抗剂BIBO 3304可阻断NPY上调α-actin作用。NPY在SAH后迟发性脑血管痉挛中发挥着重要作用。Objective To study the role of hemorrhage （SAH） in vitro cultured rat cerebral neuropeptide Y （NPY） in vasospasm after subarachnoid vascular smooth muscle cells （VSMCs） by observing the proliferation and contraction protein expression of VSMCs. Methods Male or female SD rats weighing 150 g were killed by blood loss. After infusion with saline, the circle of Willis artery and its branches were harvested and primary VSMCs were cultured. Then in vitro model of SAH was established by exposure to oxyhemoglobin of 10 s mol/L. Then the cells were treated with NPY or BIBO3304 （a commonly used blocker of NPY receptor）, or NPY -F BIBO3304. After culturing 1, 3, 5 and 7 days, immunofluorescence was used to measure the expression of ^-actin. IPP software was used to measure the fluorescence intensity and SPSS16.0 statistical software was used for data analysis. Results VSMCs were successfully cultured with digestion method. The fifth generation of visible cells showed a typical ＂valley-peak＂-like growth. In NPY and NPY -？ SAH groups, 3-day cell intervention significantly increased the fluorescence intensity of a-actin （P%0.05）, while BIBO 3304 treatment reduced the expression of a-actin, the intensity of which was as weak as that of the blank group （P〉0.05） a-actin expression in VSMCs while BIBO 3304 blocks the up-regulation of a-actin by role in delayed cerebral vasospasm after SAH. Oonclusion NPY increases NPY. NPY plays an important

关 键 词：蛛网膜下腔出血 脑血管痉挛 神经肽Y 脑血管平滑肌细胞 免疫荧光 a—actin 大鼠 

分 类 号：R743.35[医药卫生—神经病学与精神病学]