Dergunova L V, Dmitrieva V G, Filippenkov I B, Stavchansky V V, Denisova A E, Yuzhakov V V, Sevan'kaeva L E, Valieva L V, Sudarkina O Y, Gubsky L V, Myasoedov N F, Limborska S A
National Research Center "Kurchatov Institute"-Institute of Molecular Genetics, Moscow, 123182 Russia.
Pirogov Russian National Research Medical University, Moscow, Russian Federation; Federal Center of Cerebrovascular Pathology and Stroke, Ministry of Health Care of Russian Federation, Moscow, 117342 Russia.
Mol Biol (Mosk). 2021 May-Jun;55(3):402-411. doi: 10.31857/S0026898421010043.
Due to its nootropic, neuroprotective, and immunomodulatory effects, the peptide Semax is utilized in the treatment of ischemic stroke. Our earlier RNA-Seq analysis of the transcriptome in an ischemic model of transient occlusion of the middle cerebral artery showed an increase in the mRNA levels of many proinflammatory genes, and the suppression of their induction by Semax. However, for many relevant genes, including Il1a, Il1b, Il6 and Tnfa, the levels of their expression were too low for detailed quantitative evaluation. Here we utilize qRT-PCR to analyze the effects of the Semax peptide on the expression of weakly expressed mRNAs encoding several proinflammatory mediators, and show that exposure to Semax leads to a statistically significant decrease in the Il1a, Il1b, Il6, Ccl3, and Cxcl2 mRNAs, which compensates for the increase in the transcription of these genes induced by ischemia-reperfusion. We conclude that the observed protective effect of Semax in the model of stroke may be due to its anti-inflammatory effects. We also discuss the limitations of the RNA-Seq when applied to quantifying less abundant transcripts as compared to the real-time RT-PCR method.
由于具有促智、神经保护和免疫调节作用，肽类药物塞米克斯（Semax）被用于治疗缺血性中风。我们早期对大脑中动脉短暂闭塞缺血模型的转录组进行的RNA测序分析显示，许多促炎基因的mRNA水平升高，而塞米克斯可抑制这些基因的诱导表达。然而，对于许多相关基因，包括Il1a、Il1b、Il6和Tnfa，其表达水平过低，无法进行详细的定量评估。在此，我们利用qRT-PCR分析塞米克斯肽对编码几种促炎介质的低表达mRNA表达的影响，结果表明，暴露于塞米克斯会导致Il1a、Il1b、Il6、Ccl3和Cxcl2 mRNA在统计学上显著下降，这抵消了缺血再灌注诱导的这些基因转录增加。我们得出结论，塞米克斯在中风模型中观察到的保护作用可能归因于其抗炎作用。我们还讨论了与实时RT-PCR方法相比，RNA测序在定量丰度较低的转录本时的局限性。
