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Expression of heme oxygenase-1 protein and messenger RNA in permanent cerebral ischemia in rats.

机译:血红素加氧酶-1蛋白和信使RNA在大鼠永久性脑缺血中的表达

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BACKGROUND: Carbon monoxide has been regarded as a gaseous molecular messenger like nitric oxide. PURPOSE: To clarify the role of heme oxygenase-1 in the permanent cerebral ischemia at the protein and mRNA level. METHODS: The expression of heme oxygenase-1 protein and messenger RNA was investigated at different time points following MCAO using immunohistochemistry, Western blotting, RT-PCR, and Northern blotting.RESULTS: Increased HO 1immunoreactivity was detected in hippocampal and cortical neurons after 1 hour of ischemia, and was also observed in astroglial cells. After 12 hours of ischemia, HO-1 was found in both neurons and glia in cerebral cortex and thalamus, and in striatal glia cells. Western blotting analysis show the expression of HO-1 protein in cortical neurons reached the peak after 12 hours of occlusion and decreased gradually, but was still detected at day 7 post-occlusion. The expression of messenger RNA was examined in the brains of rats subjected to permanent cerebral ischemia by semi-quantitative RT-PCR and Northern blotting. HO-1 mRNA transcription could be detected 1 hour after occlusion. After 1 to 6 hours of occlusion, the expression of HO-1 rose rapidly, reaching a peak at 12 hours post-occlusion, decreased gradually, and lasted until day 7 of occlusion. Although HO activity of cerebral tissue can be detected in both sham-operated group and operated groups, the HO activity in operated groups is much stronger than that in sham-operated group. CONCLUSIONS: The induction of HO-1 protein may protect cerebral tissues from ischemic damage.
机译:背景技术:一氧化碳已被视为类似于一氧化氮的气态分子信使。目的:从蛋白和mRNA水平阐明血红素加氧酶-1在永久性脑缺血中的作用。方法:采用免疫组织化学,Western blotting,RT-PCR和Northern blotting方法研究MCAO后不同时间血红素加氧酶-1蛋白和信使RNA的表达。结果:1小时后,海马和皮层神经元HO 1的免疫反应性增加。缺血,并且在星形胶质细胞中也观察到。缺血12小时后,在大脑皮层和丘脑以及纹状体胶质细胞的神经元和神经胶质中都发现了HO-1。 Western印迹分析显示,在闭塞12小时后,皮质神经元中HO-1蛋白的表达达到峰值,并逐渐降低,但在闭塞后第7天仍检测到。通过半定量RT-PCR和Northern印迹法检测了患有永久性脑缺血的大鼠脑中信使RNA的表达。闭塞1小时后即可检测到HO-1 mRNA的转录。闭塞1至6小时后,HO-1的表达迅速上升,在闭塞后12小时达到峰值,然后逐渐下降,一直持续到闭塞第7天。尽管假手术组和手术组均可以检测到脑组织的HO活性,但假手术组的HO活性要强于假手术组。结论:HO-1蛋白的诱导可能保护脑组织免受缺血性损伤。

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