Ma Wei, Li Chun-Yan, Zhang Si-Jia, Zang Cheng-Hao, Yang Jin-Wei, Wu Zhen, Wang Guo-Dong, Liu Jie, Liu Wei, Liu Kuang-Pin, Liang Yu, Zhang Xing-Kui, Li Jun-Jun, Guo Jian-Hui, Li Li-Yan
Institute of Neuroscience, Kunming Medical University, Kunming, Yunnan Province, China.
Second Department of General Surgery, First People's Hospital of Yunnan Province, Kunming, Yunnan Province, China.
Neural Regen Res. 2022 Jun;17(6):1299-1309. doi: 10.4103/1673-5374.327346.
During acute reperfusion, the expression profiles of long noncoding RNAs in adult rats with focal cerebral ischemia undergo broad changes. However, whether long noncoding RNAs are involved in neuroprotective effects following focal ischemic stroke in rats remains unclear. In this study, RNA isolation and library preparation was performed for long noncoding RNA sequencing, followed by determining the coding potential of identified long noncoding RNAs and target gene prediction. Differential expression analysis, long noncoding RNA functional enrichment analysis, and co-expression network analysis were performed comparing ischemic rats with and without ischemic postconditioning rats. Rats were subjected to ischemic postconditioning via the brief and repeated occlusion of the middle cerebral artery or femoral artery. Quantitative real-time reverse transcription-polymerase chain reaction was used to detect the expression levels of differentially expressed long noncoding RNAs after ischemic postconditioning in a rat model of ischemic stroke. The results showed that ischemic postconditioning greatly affected the expression profile of long noncoding RNAs and mRNAs in the brains of rats that underwent ischemic stroke. The predicted target genes of some of the identified long noncoding RNAs (cis targets) were related to the cellular response to ischemia and stress, cytokine signal transduction, inflammation, and apoptosis signal transduction pathways. In addition, 15 significantly differentially expressed long noncoding RNAs were identified in the brains of rats subjected to ischemic postconditioning. Nine candidate long noncoding RNAs that may be related to ischemic postconditioning were identified by a long noncoding RNA expression profile and long noncoding RNA-mRNA co-expression network analysis. Expression levels were verified by quantitative real-time reverse transcription-polymerase chain reaction. These results suggested that the identified long noncoding RNAs may be involved in the neuroprotective effects associated with ischemic postconditioning following ischemic stroke. The experimental animal procedures were approved by the Animal Experiment Ethics Committee of Kunming Medical University (approval No. KMMU2018018) in January 2018.
在急性再灌注期间，局灶性脑缺血成年大鼠中长链非编码RNA的表达谱发生广泛变化。然而，长链非编码RNA是否参与大鼠局灶性缺血性中风后的神经保护作用仍不清楚。在本研究中，进行了RNA分离和文库制备以进行长链非编码RNA测序，随后确定已鉴定的长链非编码RNA的编码潜力并进行靶基因预测。对有或没有缺血后处理的缺血大鼠进行差异表达分析、长链非编码RNA功能富集分析和共表达网络分析。通过短暂且重复地阻断大脑中动脉或股动脉对大鼠进行缺血后处理。采用定量实时逆转录-聚合酶链反应检测缺血性中风大鼠模型中缺血后处理后差异表达的长链非编码RNA的表达水平。结果表明，缺血后处理极大地影响了缺血性中风大鼠大脑中长链非编码RNA和mRNA的表达谱。一些已鉴定的长链非编码RNA（顺式靶标）的预测靶基因与细胞对缺血和应激的反应、细胞因子信号转导、炎症和凋亡信号转导途径有关。此外，在接受缺血后处理的大鼠大脑中鉴定出15种显著差异表达的长链非编码RNA。通过长链非编码RNA表达谱和长链非编码RNA-mRNA共表达网络分析鉴定出9种可能与缺血后处理相关的候选长链非编码RNA。通过定量实时逆转录-聚合酶链反应验证表达水平。这些结果表明，已鉴定的长链非编码RNA可能参与缺血性中风后与缺血后处理相关的神经保护作用。实验动物程序于2018年1月获得昆明医科大学动物实验伦理委员会批准（批准号：KMMU2018018）。
