Key Laboratory of Critical Disease Mechanism and intervention of Hebei Province, Department of Pathophysiology, Hebei Medical University, Shijiazhuang, China.
Key Laboratory of Medical Biotechnology of Hebei Province, Hebei Medical University, Department of Biochemistry and Molecular Biology, Cardiovascular Medical Science Center, Shijiazhuang, China.
Mol Neurobiol. 2022 Jul;59(7):3996-4014. doi: 10.1007/s12035-022-02841-3. Epub 2022 Apr 22.
Long noncoding RNAs (lncRNAs) play an important regulatory role in various diseases. However, the role of lncRNAs in brain ischemic tolerance (BIT) induced by cerebral ischemic preconditioning (CIPC) is still unknown. The lncRNA profile of rat cortical astrocytes pretreated with ischemic preconditioning was analyzed by high-throughput sequencing. The results of Cell-Counting Kit-8 (CCK-8) assay showed that a novel lncRNA, NONRATT009133.2, which we referred to as brain ischemia-related factor (BIRF), was highly correlated with BIT. Through bioinformatics analysis, we predicted that BIRF, miR-330-5p, and GLT-1 (also named Slc1a2) might constitute a ceRNA regulatory network in the induction of BIT. We found that BIRF was upregulated by CIPC, which promoted GLT-1 expression and BIT induction. BIRF could directly bind to miR-330-5p. Furthermore, miR-330-5p directly targeted GLT-1, and miR-330-5p inhibited both GLT-1 expression and BIT induction in vitro and in vivo. Moreover, BIRF acts as a molecular sponge to competitively bind to miR-330-5p with GLT-1 mRNA, while the miR-330-5p inhibitor reversed all the effects of BIRF siRNA on GLT-1 expression and neuronal vitality. Taken together, our results demonstrate the important roles of the BIRF/miR-330-5p/GLT-1 axis in the induction of BIT by CIPC. BIRF may be a potentially effective therapeutic strategy against stroke injury.
长链非编码 RNA（lncRNA）在各种疾病中发挥重要的调控作用。然而，lncRNA 在脑缺血预处理（CIPC）诱导的脑缺血耐受（BIT）中的作用尚不清楚。通过高通量测序分析预处理缺血的大鼠皮质星形胶质细胞的 lncRNA 图谱。Cell-Counting Kit-8（CCK-8）检测结果表明，一种新的 lncRNA，NONRATT009133.2，我们称之为脑缺血相关因子（BIRF），与 BIT 高度相关。通过生物信息学分析，我们预测 BIRF、miR-330-5p 和 GLT-1（也称为 Slc1a2）可能构成诱导 BIT 的 ceRNA 调控网络。我们发现 CIPC 上调 BIRF，促进 GLT-1 表达和 BIT 诱导。BIRF 可直接与 miR-330-5p 结合。此外，miR-330-5p 直接靶向 GLT-1，miR-330-5p 抑制体外和体内 GLT-1 表达和 BIT 诱导。此外，BIRF 作为分子海绵与 GLT-1 mRNA 竞争性结合 miR-330-5p，而 miR-330-5p 抑制剂逆转了 BIRF siRNA 对 GLT-1 表达和神经元活力的所有影响。总之，我们的结果表明 BIRF/miR-330-5p/GLT-1 轴在 CIPC 诱导 BIT 中的重要作用。BIRF 可能是一种针对中风损伤的潜在有效治疗策略。
