胰岛素；脑缺血；Bcl2_

脑室内注射胰岛素对大鼠全脑缺血后海马CA1区Bcl2 mRNA变化及其蛋白表达的影响

http://www.100md.com 《中华首席医学网》 2006年第3期

胰岛素；脑缺血；Bcl2,,胰岛素；脑缺血；Bcl2,脑室内注射胰岛素对大鼠全脑缺血后海马CA1区Bcl2mRNA变化及其蛋白表达的影响,摘要,关键词,1材料与方法,2结果,3讨论,参考文

     摘要：目的观察胰岛素对全脑缺血后海马CA1区Bcl2 mRNA含量变化及其蛋白表达的影响，探讨胰岛素保护作用的机制。方法利用4VO法制作大鼠全脑缺血模型。造成脑缺血15min后行再灌注，治疗组于再灌注后即刻经脑室注入1u胰岛素(10μL，1×105u/L)，缺血组则经脑室注入10μL生理盐水。利用反转录PCR(RTPCR)、免疫组化及Westernblot法分别于全脑缺血后1、3、5d观察海马CA1区Bcl2蛋白表达及其mRNA相对含量的变化。结果全脑缺血后1、3、5d治疗组海马CA1区可见呈阳性表达的Bcl2蛋白，而缺血组海马CA1区Bcl2蛋白的表达则呈阴性；Westernblot分析，全脑缺血后1、3、5d治疗组海马CA1区Bcl2蛋白电泳条带密度分别为2890.791±213.616，3147.396±243.561和2594.834±204.736，明显高于缺血组的743.376±84.123，804.637±64.547和637.867±54.394(P<0.01)。全脑缺血后1、3、5d缺血组及胰岛素治疗组海马CA1区Bcl2 mRNA相对含量则未见明显差异。结论全脑缺血后脑室内注入胰岛素可能通过促进海马CA1区神经元Bcl2基因的翻译途径从而促进Bcl2蛋白表达，这可能是其对全脑缺血后海马CA1区神经元产生中枢直接保护作用的主要机制之一。

    关键词：胰岛素；脑缺血；Bcl2

    The effect of intraventricular administration of insulin on expression of Bcl2 mRNA and protein in rat hippocampal CA1 region following global brain ischemia

    Li Bing, Zhang Xiang, Jiang Xiaofan, Wang Yangang, Zhang Ge

    (Department of Neurosurgery, Xijing Hospital of Fourth Military Medical University, Xian 710032, China)

    ABSTRACT: Objective To investigate the mechanism of neuroprotective effect of insulin on hippocampal CA1 region, the relatively content of Bcl2 mRNA and the expression of Bcl2 protein in rat hippocampal CA1 region following global brain ischemia were observed. Methods Fourvessel occlusion was used to establish the rat global brain ischemic model. After 15 minutes brain ischemia, 1 unit insulin (10μL, 1×105u/L) was injected into the cerebral ventricular immediately post reperfusion in treated group, while, 10μL NaCl (90g/L) solution was injected into the cerebral ventricular at the same time in ischemic group. The relatively content of Bcl2 mRNA and the expression of Bcl2 protein in hippocampal CA1 region had been detected by reverse transcription polymerase chain reaction (RTPCR), immunocytochemistry, and Westernblot, respectively. Results The expression of Bcl2 protein was negative in ischemic group and was positive in treated group, in 1, 3, 5 day post brain ischemia. While, Westernblotting analysis showed that the density of electrophoretic strip of Bcl2 protein was 2890.791±213.616, 3147.396±243.561 and 2594.834±204.736 in treated group. It was much higher than that in ischemic group, which was 743.376±84.123, 804.637±64.547 and 637.867±54.394, respectively. But, the relatively content of Bcl2 mRNA had no obvious difference between ischemic group and treated group. Conclusion Intraventricular administration of insulin can improve the expression of Bcl2 protein by up regulating the translation of the Bcl2, and this may be the main point to achieve the neuroprotective effect on hippocamal CA1 region post global brain ischemia. ......

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