【摘要】 目的:观察局灶性脑缺血再灌注信号转导与转录激活子1蛋白表达及其丝氨酸残基磷酸化,探讨其在神经元凋亡中的作用。方法:实验于2003-04/2004-01在中南大学湘雅医院神经病学研究所和中心实验室完成。选择雄性SD大鼠52只,随机分为空白对照组4只,假手术组16只,脑缺血组32只,脑缺血组又分为再灌注15min,1,4,12,24,48h,5,7d8个时相点,每个时相点4只。采用线栓法制作短暂性局限脑缺血模型。造模后进行神经功能评分,0分:正常,未见任何神经功能缺损表现。1分:垂直提起时前爪不能伸直。2分:行走时身体向左倾,向左侧旋转。3分:行走时身体向左侧跌倒。4分:不能自发行走或有意识障碍。根据首次评分,没有神经功能缺损、评分4分、有呼吸困难、提前死亡及处死时发现有蛛网膜下腔出血的动物均弃去。此外对于手术中出血过多的动物也弃去。采用免疫组织化学技术依靠特异性抗体检测信号转导与转录激活子1蛋白表达及其丝氨酸残基磷酸化。结果:纳入动物52只,均进入结果分析。①信号转导与转录激活子1于再灌注1h检测到阳性表达,12h达到峰值,24h下降[分别为(16.90±2.95),(28.40±4.62),(18.68±2.72)个]。②信号转导与转录激活子1磷酸化于再灌注15min出现,4h达到峰值,持续至少5d[分别为(7.87±1.82),(28.43±5.94),(10.25±2.48)个]。结论:脑缺血再灌注信号转导与转录激活子1表达上调并在丝氨酸残基磷酸化,可能参与再灌注神经元凋亡。更多还原

【Abstract】 AIM: To observe the expression and serine phosphorylation of signal transduction and activator of transcription 1 (STAT1) followed focal cerebral ischemic reperfusion, and explore its role in neuronal apoptosis. METHODS: The experiment was conducted in the Institute of Neurology and the Central Laboratory of Xiangya Hospital, Central South University from April 2003 to January 2004. Fifty-two male SD rats were randomly divided into control group (n=4), sham operation group (n=16) and cerebral ischemia group (n=32). The cerebral ischemia group was subdivided into eight groups according to eight different reperfusion phases: 15 minutes, 1 hour, 4, 12, 24 hours, 5 and 7 days after reperfusion with 4 rats in each group. The suture method was applied to prepare the transitory focal cerebral ischemic rat model. The neurologic evaluation was performed after establishment of model: 0 meant normal, no neurologic defects; 1 point: failure to extend left forepaw fully; 2 points: circling to the left when walking; 3 points: falling to the left when walking; 4 points: disable to walk spontaneously or having a consciousness disorder. The animals with no neurologic defects, 4 scores of evaluation, dyspnea, advanced death or subarachnoid hemorrhage when killed were excluded after the first evaluation. Additionally, the animals with overmuch hemorrhage when operated were excluded too. The expression and serine phosphorylation of STAT1 were detected by immunohistochemistry with specific antibodies. RESULTS: All 52 rats entered the result analysis. ①STAT1 positive cells were firstly detected at 1 hour after reperfusion, reached peak at 12 hours, and declined after 24 hours [(16.90±2.95), (28.40±4.62), (18.68±2.72) cells, respectively]. ②The serine phosphorylation of STAT1 was firstly detected at 15 minutes after reperfusion, reached the peak at 4 hours, and lasted at least 5 days [(7.87±1.82), (28.43±5.94), (10.25±2.48) cells, respectively]. CONCLUSION:The increased expression and serine phosphorylation of STAT1 followed focal cerebral ischemic reperfusion may account for the neuronal apoptosis.更多还原