脑血管痉挛的差异蛋白质组学研究

目的采用蛋白质组学技术分离和鉴定动脉瘤性蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)患者组及无痉挛患者组发病后第1 d脑脊液中差异表达蛋白质,寻找与脑血管痉挛相关的蛋白质。方法采用双向凝胶电泳分离无脑血管痉挛组(对照组)、轻中度痉挛组(实验组1)和重度痉挛组(实验组2)脑脊液总蛋白质,比较分析实验组和对照组之间的差异表达蛋白质,应用基质辅助激光解析电离串联飞行时间质谱(MALDI-TOF/TOF-MS)鉴定差异表达的蛋白质。结果实验组1、实验组2与对照组表达差异2倍以上的蛋白质斑点共有68个,成功鉴定出23个差异表达蛋白质。结论初步建立了脑血管痉挛的差异表达蛋白质双向电泳技术体系,筛选出一批与血管痉挛相关蛋白质候选物。本研究为脑血管痉挛的发病机制研究提供了新的方法,为寻找脑血管痉挛的生物标志物、寻找药物治疗靶标和预后评估打下研究基础。     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

颅脑创伤后亚低温脑保护的蛋白质组研究

目的 本研究应用差异蛋白质组学技术,探讨亚低温和常温条件下,创伤性颅脑损伤大鼠海马组织蛋白质的表达变化.方法 采用侧方液压冲击装置,建立大鼠中度脑损伤模型,亚低温组(n=3)于伤后维持体温(33±0.5)℃持续3h,常温组(n=3)始终维持体温(37±0.5)℃.取大鼠海马组织,通过差异荧光双向凝胶电泳、分离蛋白,获得二维的蛋白质分离图谱,然后通过胶内酶切、抽提酶解肽段、基质辅助激光解吸飞行时间质谱分析差异的蛋白质点,鉴定出变化的蛋白质.结果 通过DeCyder5.0(GE Healthcare)软件分析报告发现差异1.5倍以上的蛋白点17个(P＜0.05).通过对这些蛋白考染点进行质谱鉴定,鉴定出14个蛋白质,2个为同一种蛋白,实际差异蛋白数为13个.分别是细胞骨架蛋白、介导能量代谢的酶类、参与核酸合成、氧化应激反应的蛋白质、神经突触功能蛋白、细胞内信号传递蛋白及未知蛋白.结论 脑损伤后亚低温及常温条件下,大鼠海马组织蛋白质存在表达差异,鉴定出的差异蛋白质可能与亚低温保护效应的潜在作用机制有关.     

侵袭性与非侵袭性垂体腺瘤的蛋白质差异表达

目的 建立分辨率高和重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出其差异表达的蛋白质.方法 利用固相pH梯度双向凝胶电泳分离侵袭性与非侵袭性垂体腺瘤的总蛋白质,凝胶经银染显色后,运用ImageMaster 2D图像分析软件进行比较分析、识别差异表达的蛋白质,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)及数据库搜索鉴定部分差异蛋白质点.结果 得到了分辨率较高、重复性好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱.比较分析图谱,初步对其中30个点进行了肽质指纹图分析,鉴定出一些与细胞周期调控、信号传导等有关的差异蛋白质.结论 建立了分辨率较高且重复性较好的侵袭性与非侵袭性垂体腺瘤的双向凝胶电泳图谱,并识别鉴定出一些侵袭性与非侵袭性垂体腺瘤的差异表达的蛋白质,为进一步筛选侵袭性垂体腺瘤的蛋白质表达数据库提供了基础.     

缝隙连接阻断剂1-庚醇对脑血管痉挛的抑制作用

目的探讨缝隙连接在脑血管痉挛中的作用及观察其阻断剂在动物实验中的治疗作用。方法建立兔二次蛛网膜下腔出血模型,通过脑血管造影观察经动脉或池内注入缝隙连接阻断剂heptanol,对脑血管痉挛的抑制和治疗作用,并观察基底动脉的形态学变化。结果枕大池注血后血管造影显示基底动脉出现痉挛。在脑血管痉挛后,动脉给予heptanol对急、慢性脑血管痉挛有显著的治疗作用。预先枕大池注入heptanol再注血,造影显示基底动脉痉挛不明显(P>0.05),heptanol枕大池预处理后能显著抑制急、慢性期脑血管痉挛的形成。形态学检查发现,对照组第7d的基底动脉光镜见内皮细胞核染色质聚集,内弹力膜波纹状,平滑肌细胞分布稀疏等。动脉给药组和池内给药组也有类似变化,但范围局限、程度轻。结论缝隙连接阻断剂heptanol能有效抑制兔蛛网膜下腔出血后急性和慢性脑血管痉挛,体内试验表明缝隙连接在脑血管痉挛中可能发挥重要作用,应用其阻断剂heptanol具有显著的治疗作用。     

Role of Ca(2+)-dependent K+ channels in erythrocyte lysate-induced contraction of rabbit cerebral artery.

K+ channel openers may be useful in the treatment of cerebral vasospasm following subarachnoid hemorrhage. However, the role of Ca(2+)-dependent K+ channel (KCa) openers in cerebral vasospasm remain unclear. This study was undertaken to examine the role of KCa in hemolysate-induced contraction of rabbit cerebral and peripheral arteries: 1. Iberiotoxin (IBX), a selective KCa channel blocker, produced more pronounced contraction in basilar than in those of carotid or femoral arteries, indicating KCa channels are important regulating factors in cerebral arteries; 2. NS1619, a selective KCa channel opener, abolished the contraction of basilar artery to erythrocyte lysate, a causative agent for cerebral vasospasm; 3. In rabbit basilar artery, NS1619 relaxed the contractions to IBX, erythrocyte lysate and KCl (20 and 60 mM), indicating that NS1619, besides opening KCa channels, possesses other vasodilating actions. We conclude that KCa channels are important factors in the regulation of cerebral vascular tension and KCa channel opener NS1619 may have dual relaxant actions in cerebral arteries.     

颅内动脉瘤患者血清差异表达蛋白质的研究

目的 采用蛋白质组学方法研究散发性颅内动脉瘤和健康成人外周血清蛋白表达差异.方法 对散发性颅内动脉瘤和健康成人的血清样本进行双向凝胶电泳分离,通过对两种蛋白图谱的比较,确定差异表达的蛋白点;而后对差异点进行基质辅助激光解析电离飞行时间质谱分析和蛋白数据库信息检索;随机选取2种蛋白进行Western blot法验证蛋白质组学实验结果.结果 在颅内动脉瘤和健康成人中鉴定出14种差异表达蛋白,得到了11个肽质量指纹图谱(peptide mass fingerprinting,PMF),最终7种蛋白质得到确认.结合珠蛋白、C反应蛋白、B因子、淀粉样血清蛋白A和载脂蛋白E前体在动脉瘤组表达上调;载脂蛋白A-Ⅳ前体和间α胰蛋白酶抑制物H4重链前体表达下调.蛋白印迹实验结果与双向电泳结果一致.结论 成功鉴定出7个与颅内动脉瘤相关的蛋白,为进一步阐明颅内动脉瘤形成的病理生理学机制提供了新的思路.     

Role of Ca2+-dependent K+ channels in erythrocyte lysate-induced contraction of rabbit cerebral artery

Abstract

K⁺ channel openers may be useful in the treatment of cerebral vasospasm following subarachnoid hemorrhage. However, the role of Ca²⁺ -dependent K⁺ channel (KCa) openers in cerebral vasospasm remain unclear. This study was undertaken to examine the role ofKCa in hemolysate-induced contraction of rabbit cerebral and peripheral arteries: 7. Iberiotoxin (IBX), a selective KCa channel blocker, produced more pronounced contraction in basilar than in those of carotid or femoral arteries, indicating KCa channels are important regulating factors in cerebral arteries; 2. NS1619, a selective KCa channel opener, abolished the contraction of basilar artery to erythrocyte lysate, a causative agent for cerebral vasospasm; 3. In rabbit basilar artery, NS1619 relaxed the contractions to IBX, erythrocyte lysate and KCI (20 and 60 mM), indicating that NS1619, besides opening KCa channels, possesses other vasodilating actions. We conclude that KCa channels are important factors in the regulation of cerebral vascular tension and KCa channel opener NS1619 may have dual relaxant actions in cerebral arteries. [Neurol Res 1999; 21: 705–711 ]     

Therapeutic potential of peroxisome proliferator-activated receptor γ agonist rosiglitazone in cerebral vasospasm after a rat experimental subarachnoid hemorrhage model

The pathogenesis of cerebral vasospasm is closely associated with inflammation and immune response in arterial walls. Recently, the authors proved the key role of Toll-like receptor (TLR)4 in the development of vasospasm in experimental subarachnoid hemorrhage (SAH) model. Because peroxisome proliferator-activated receptor (PPAR) gamma agonists are identified as effective inhibitors of TLR4 activation, we investigated the anti-inflammation properties of PPAR-gamma agonist rosiglitazone in basilar arteries in a rat experimental SAH model and evaluated the effects of rosiglitazone on vasospasm. Inflammatory responses in basilar arteries were assessed by immunohistochemical staining for intercellular molecule (ICAM)-1 and myeloperoxidase (MPO). Expression of TLR4 was determined by western blot analysis. The degree of cerebral vasospasm was evaluated by measuring the mean diameter and cross-sectional area of basilar arteries. Rosiglitazone suppressed the SAH-induced inflammatory responses in basilar arteries by inhibiting the TLR4 signalling. Furthermore, rosiglitazone could attenuate cerebral vasospasm following SAH. Therefore, we suggested that PPAR-gamma agonists may be potential therapeutic agents for cerebral vasospasm.     

Potential role of Ras in cerebral vasospasm after experimental subarachnoid hemorrhage in rabbits

Previous studies have demonstrated that mitogen-activated protein kinase (MAPK) is involved in the pathogenesis of cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH). Ras, an upstream regulator of MAPK, may be activated following SAH. The aim of this study was to investigate the role of Ras in cerebral vasospasm in a rabbit model of SAH. We first investigated the time course of Ras and ERK1/2 activation in the basilar artery after SAH. Next, for the time point at which Ras was maximally activated, we assessed the effect of FTI-277 (a Ras farnesyltransferase inhibitor) on cerebral vasospasm. SAH was induced by injecting autologous blood into the cisterna magna on both day 0 and day 2. FTI-277 was injected into the cisterna magna every 24 hours, beginning 30 minutes after blood injection to the last day of the experiment. Elevated expression of Ras-GTP and phosphorylated ERK1/2 was detected in the basilar artery after SAH and expression peaked on day 3. FTI-277 administration resulted in lower Ras-GTP and phosphorylated ERK1/2 levels and markedly attenuated vasospasm in the basilar arteries relative to animals that did not receive FTI-277. Our results suggest that Ras protein is activated in the arterial wall after SAH and contributes to vasospasm development.     

Cerebral vasospasm: comparison of contractile responses in isolated human and canine basilar arteries

Differences between human and canine basilar arteries in contractile responses to various agents were studied in vitro. Human basilar arteries were obtained at postmortem. Serotonin or prostaglandin F2 alpha contracted greatly both human and canine basilar arteries. There was no difference between the two vessels in serotonin- or prostaglandin F2 alpha-induced contractions. In contrast, great differences were found between the two arteries in response to norepinephrine or hemoglobin. Human basilar arteries contracted markedly to norepinephrine while canine basilar arteries did not contract to norepinephrine. This data suggests that sympathetics might play a more important role in the genesis of vasospasm after subarachnoid hemorrhage than has been previously thought. Hemoglobin did not elicit a contraction in human basilar arteries whereas it produced marked contractions in canine basilar arteries. This result indicates that possible participation of hemoglobin as a causative agent in vasospasm could be ruled out. In view of these differences between human and canine cerebral arteries, the canine seems unsuitable for an experimental model of cerebral vasospasm.     

缝隙连接蛋白43在内皮素诱导的脑基底动脉收缩中的表达变化

目的 探讨Cx43在内皮素诱导的脑基底动脉收缩中的表达变化及其可能的作用.方法 血管环张力实验检测内皮素诱导的脑基底动脉的收缩变化并应用Western blot检测基底动脉Cx43蛋白的表达变化,染料传输实验用来检测脑基底动脉收缩过程中平滑肌细胞间缝隙连接的功能变化.结果 浓度递增的内皮素导致脑基底动脉呈显著浓度依赖性的收缩,一定浓度缝隙连接阻断剂苷珀酸显著缓解该收缩;收缩过程中,Cx43的蛋白表达呈显著时间依赖性的升高,苷珀酸减弱该表达的升高;内皮素刺激下,血管平滑肌细胞间的染料传输呈时间依赖性的升高,苷珀酸显著减少染料在细胞间的传输.结论 脑血管痉挛过程中,通过增加Cx43的表达,血管细胞间缝隙连接的功能被内皮素激活并在血管痉挛病理过程中发挥重要作用;抑制缝隙连接的功能是有效缓解蛛网膜下腔出血后脑血管痉挛的新途径.     

实验性蛛网膜下腔出血后脑血管痉挛兔基底动脉Cx43蛋白表达的时相变化

目的 通过建立兔二次蛛网膜下腔出血实验模型,观察兔蛛网膜下腔出血后基底动脉缝隙连接蛋白Cx43表达的时相变化特点,初步探讨蛛网膜下腔出血后脑血管痉挛的形成机制.方法 选择健康新西兰大白兔30只,随机分为5组:正常对照组(n=6)和蛛网膜下腔出血模型组(1d、3d、7d和14d,n=6):建立兔二次蛛网膜下腔出血后脑血管痉挛实验模型,脑血管造影分析基底动脉的直径变化并应用Western Blot检测基底动脉Cx43蛋白的表达变化.对血管直径与Cx43表达变化情况进行相关分析.结果 成功建立兔二次蛛网膜下腔出血模型;脑血管造影显示注血后1d基底动脉即出现痉挛(85.7%±8.6%,P<0.05);7d时达高峰(66.5%±7.6%,P<0.01);14d时仍有痉挛(78.4%±8.2%,P<0.05)但程度较前缓解.Cx43蛋白表达在建立SAH模型后1d(38.6%±5.6%,P<0.05)、3d(50.2%±5.7%,P<0.05)、7d(57.8%±5.3%,P<0.01)、14d(32.4%±3.6%.P<00.05)均升高,其中7d为高峰,14d开始下降.Cx43蛋白表达的时相性变化与SAH后基底动脉直径的时相性变化相关系数为0.914.结论 实验结果 显示蛛网膜下腔出血后兔基底动脉缝隙连接蛋白Cx43的表达发生了时相性变化,并且Cx43蛋白表达强弱与蛛网膜下腔出血后脑血管痉挛程度在时程上存在正相关关系,表明缝隙连接蛋白Cx43可能参与蛛网膜下腔出血后脑血管痉挛的形成.

Abstract：

Objective The study was designed to explore the change of expression of connexin43(Cx43)protien in the model of subarachnoid hemorrhage(SAH)of rabbits,hoping to provide the basis to study the mechanism of cerebral vasospasm(CVS).Methods 30 New Zealand rabbits were divided into 5 groups:SAH group(1d,3d,7d,14d,n=6)and control group(n=6).The model of CVS following SAH was established.Digital subtraction angiography was performed to detect the change of the basilar arteries diameter.The expression of Cx43 protien in basilar arteries tissue at different time points following experimental SAH was examined by using western blotting analysis.The data were statistically analyzed using the bivariate correlations test.Results The model of SAH in rabbits was successfully established.All 30 rabbits were analyzed.Cerebral angiograms on 1d,3d,7d and 14d showed severe narrowing of the BAs,and on 7d showed the most narrowing and on 14d began to Relieve.Western blotting showed that the expression of Cx43 protein were detected in normal rabbit basilar arteries tissue.However,the expression of Cx43 protein increased gradually and significantly in models compared with that of control(P<0.05),which reached peak on 7d(P<0.01)and then decreased on 14d(P<0.05).There was positive correlation between expression of Cx43 and cerebral vasospasm.Conclusions The above results demonstrates at the first time that the Cx43 protein expression is altered after the SAH,and exhibits a time-dependent change.which might be connected with the development of CVS.In summary,our data demonstrates gap junctions may play an important role in the pathogenesis of cerebral vasospasm after SAH.     

甘珀酸对兔脑血管痉挛时缝隙连接蛋白43磷酸化表达的影响

目的 探讨甘珀酸对兔实验性蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)时缝隙连接蛋白43(Cx43)磷酸化表达的影响.方法 建立兔二次SAH模型,脑池及静脉分别给予甘珀酸,脑血管造影及光镜观察分析基底动脉直径及形态学变化并应用Western blot检测基底动脉Cx43蛋白磷酸化表达的变化.结果 SAH组与正常组相比,脑血管造影及光镜观察结果 证实基底动脉痉挛明显;痉挛动脉肇磷酸化的Cx43(P-Cx43)蛋白表达显著升高,但去磷酸化的Cx43(NP-Cx43)蛋白表达显著减少.甘珀酸脑池处理组及静脉处理组与SAH组相比,脑血管造影及光镜观察结果 证实基底动脉痉挛显著减轻;痉挛动脉壁P-Cx43蛋白表达显著减少,但NP-Cx43蛋白表达显著升高.结论 SAH后,Cx43蛋白磷酸化表达发生变化,脑池或静脉给予甘珀酸能明显缓解SAH后CVS,其作用机制可能与基底动脉Cx43蛋白磷酸化表达变化有关.