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背景:脑缺血-再灌注损伤与 Kinesin 活性密切相关,研究认为微管运动蛋白 Kinesin 活性下降是脑缺血后神经细胞死亡的早期标志。黄芩素甙可以防止脑缺血-再灌注诱发的蛋白激酶 C 的激活、减轻钙超载,且可减小缺血梗死灶的体积,从而减轻脑缺血-再灌注损伤。但是黄芩素甙脑保护作用是否与 Kinesin 活性有关,目前很少报道。目的:观察黄芩素甙对脑缺血-再灌注期间海马锥体细胞微管运动蛋白Kinesin 活性的变化。设计:随机对照实验。单位:济宁医学院附属医院麻醉科及徐州医学院附属医院江苏省麻醉学重点实验室。材料:实验于 1999-02/08在江苏省麻醉学重点实验室完成。选用雄性沙土鼠 35只。方法:将沙土鼠随机分成假手术组(5只)、缺血再灌注对照组(15只)和黄芩素甙组(15只)3组,根据再灌注时间将缺血再灌注对照组和黄芩素甙组分为 3个亚组,即再灌注Ⅰ组(再灌注 6h)、再灌注Ⅱ组(再灌注 48h)和再灌注Ⅲ组(再灌注 96h)。每亚组 5只动物。缺血再灌注对照组和黄芩素甙组制备沙土鼠前脑缺血-再灌注损伤模型,脑缺血时间为 10m in。假手术组仅游离双侧颈总动脉但不予阻断。黄芩素甙组于缺血前 15m in 给予灯盏花素(其有效成分为黄芩素甙)45m g/kg腹腔注射,假手术组和缺血再灌注对照组则给予等量的生理盐水灌胃。应用免疫组织化学染色方法结合计算机图象分析技术测定海马微管运动蛋白 Kinesin 的活性。主要观察指标:各组动物 Kinesin 的活性及其变化。结果:纳入实验的动物为 35只,均进入结果分析,无实验动物脱失。在海马 CA1区,缺血再灌注对照组缺血-再灌注 6,48和 96h 时微管运动蛋白 Kinesin 活性分别降至假手术组的 58%,38%和 12%(P 均<0.01),而黄芩素甙组在再灌注 6,48和 96h 后,微管运动蛋白 Kinesin 活性分别为假手术组的 81%,61%和 21%,均明显高于缺血再灌注对照组(P均<0.05)。在海马 CA2,CA3和 CA4区,微管运动蛋白 Kinesin 的活性无明显变化。结论:黄芩素甙可通过抑制脑缺血-再灌注期间海马 CA1区微管运动蛋白 Kinesin 活性的下降达到脑保护作用。
Background: Cerebral ischemia-reperfusion injury is closely related to Kinesin activity. Studies suggest that decreased Kinesin activity of microtubule motor protein is an early marker of neuronal cell death after cerebral ischemia. Safrole can prevent the activation of protein kinase C induced by cerebral ischemia-reperfusion, reduce calcium overload, and reduce the volume of ischemic infarcts, thereby reducing cerebral ischemia-reperfusion injury. However, whether the safrole protective effect of safrole is related to Kinesin activity is rarely reported. OBJECTIVE: To observe the changes of microtubule kinesin Kinase activity in hippocampal pyramidal neurons induced by baicalein during cerebral ischemia-reperfusion. Design: Randomized controlled trials. Unit: Department of Anesthesiology, Affiliated Hospital of Jining Medical College; Key Laboratory of Anesthesiology of Jiangsu Province, Affiliated Hospital of Xuzhou Medical College. MATERIALS: The experiment was performed at the Key Laboratory of Anesthesiology of Jiangsu Province from February to August 1999. Thirty-five male gerbils were used. METHODS: Gerbils were randomly divided into sham-operated group (5 rats), ischemia-reperfusion control group (15 rats), and baicalein-treated group (15 rats). According to the time of reperfusion, ischemia-reperfusion control group and Astragalus membranaceus were used. The quinone component was divided into 3 subgroups, ie reperfusion group I (reperfusion 6h), reperfusion group II (reperfusion 48h) and reperfusion group III (96h reperfusion). 5 animals per subgroup. Cerebral ischemia-reperfusion injury model was established in the gerbil ischemia-reperfusion control group and baicalein treatment group. The cerebral ischemia time was 10m in. The sham group only dissociated the common carotid artery but did not block it. In the Astragalus membranaceus group, breviscapine (available as baicalein) was intraperitoneally injected at 15m g/kg 15m before ischemia, and sham-operated and ischemia-reperfusion control groups were given intragastric administration of equal volume of normal saline. The activity of Kinesin, a hippocampal microtubule motor protein, was determined by immunohistochemical staining and computer image analysis. MAIN OUTCOME MEASURES: Kinesin activity and changes in various groups of animals. RESULTS: Thirty-five animals were included in the experiment and all were involved in the analysis of the results. No experimental animal was lost. In hippocampal CA1 region, the activity of Kinesin Kinesin decreased to 58%, 38%, and 12% in the sham-operated group at 6, 48, and 96 hours after ischemia-reperfusion in the ischemia-reperfusion control group, respectively (P < 0.01). The activity of Kinesin in the sham operation group was 81%, 61%, and 21% in the sham-operated group, which were significantly higher than those in the ischemic-reperfusion control group (P<0.01) after 8, 48, and 96 hours of reperfusion. 0.05). In the CA2, CA3 and CA4 regions of the hippocampus, there was no significant change in the activity of the microtubule motor Kinesin. CONCLUSION: Baicalein can achieve brain protection by inhibiting the decrease of Kinesin activity of hippocampal CA1 region during cerebral ischemia-reperfusion.