Servicio de Bioquímica-Investigación, Hospital Ramón y Cajal, Irycis, Madrid, Spain.
Neurochem Res. 2010 Aug;35(8):1239-47. doi: 10.1007/s11064-010-0180-9. Epub 2010 May 11.
An antibody microarray was used to analyze potential modifications in brain protein levels induced by ischemic reperfusion. Total brain extracts from rats subjected to 15 min of transient global ischemia followed by 3 days of reperfusion and sham control animals were compared within the same array. Separate arrays were run to analyze resistant (cortex) and vulnerable (CA1) regions to ischemia. Candidate components distinguishing the two cellular populations were selected under stringent criteria. IR significantly decreased the expression of Bcl-x, caspase 11, GADD153, Cdk4, E2F1, Retinoblastoma-P, SMAD4, AP-1/c-jun, ATF2, PCAF, MAP1b and cofilin within both regions. NGF and NMDA 2A receptors and IkappaB were specifically down-regulated in CA1, while Pyk2-P, b-NOS, and tyrosine hydroxylase were slightly up-regulated in the same region. Some of the array results were validated by western blot. Both the array and western blot results suggested a relevant IR induced activation of calpain specifically at CA1.
采用抗体微阵列分析了脑缺血再灌注诱导的潜在蛋白水平变化。在同一芯片上比较了短暂全脑缺血后 3 天再灌注和假手术对照大鼠的全脑提取物。分别运行数组来分析耐缺血（皮质）和易损（CA1）区。在严格的标准下选择区分两个细胞群体的候选成分。IR 显著降低了两个区域中 Bcl-x、caspase 11、GADD153、Cdk4、E2F1、Retinoblastoma-P、SMAD4、AP-1/c-jun、ATF2、PCAF、MAP1b 和肌动蛋白的表达。NGF 和 NMDA 2A 受体和 IkappaB 在 CA1 中特异性下调，而 Pyk2-P、b-NOS 和酪氨酸羟化酶在同一区域略有上调。一些阵列结果通过 Western blot 进行了验证。阵列和 Western blot 结果均提示 CA1 中钙蛋白酶的激活。
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