Analysis of peptide Semax regulatory action with help of RNA-Seq analysis of the model of transient focal ischemia in rat brain

The model of transient cerebral focal ischemia was used. It was realized on two-month-old male white rats of Wistar line. Animals were divided into two groups: "ischemia+Semax". Focal ischemia in experimental rats was initiated by endovascular occlusion of right middle cerebral artery using monofilament for 90 minutes followed by reperfusion (tMCAO). Neuropeptide Semax was administered intraperitoneally at a dose of 10 μg / 100 g of the rat weight at 90 min, as well as 2.5 and 6.5 hours after tMCAO. Rats were withdrawn from the experiment 4.5 and 24 hours after the occlusion. Each group contained 3 animals. Ischemic focus was localized in the cerebral subcortex. Genome-wide transcriptome analysis of mRNAs level in rat subcortex was carried out using High-throughput RNA-sequencing (RNA-Seq). PolyA fraction from total RNA was used for RNA-seq. RNA-seq was carried out using Illumina HiSeq1500 instrument (reads 50 nt).

本研究采用短暂性局灶性脑缺血模型，实验对象为2月龄雄性Wistar系白化大鼠。将实验动物分为两组：「缺血+塞美克斯（Semax）」组。实验组大鼠通过线栓法阻塞右侧大脑中动脉90分钟后再灌注，构建线栓法大脑中动脉阻塞再灌注（tMCAO）模型。分别于脑缺血再灌注后90分钟、2.5小时及6.5小时，按10μg/100g大鼠体重的剂量，对实验组大鼠腹腔注射神经肽塞美克斯（Semax）。分别于动脉阻塞后4.5小时及24小时终止实验，每组各包含3只大鼠。缺血灶定位于大脑皮层下区域。本研究采用高通量RNA测序（High-throughput RNA-sequencing，RNA-Seq）对大鼠皮层下组织的mRNA水平进行全基因组转录组分析，以总RNA中的PolyA富集片段作为建库材料，测序工作通过Illumina HiSeq1500平台完成，读取长度为50nt。