Department of Emergency Medicine, The Second Affiliated Hospital, Zhejiang University School of Medicine.
Department of Emergency Medicine, Yuhang Branch of the Second Affiliated Hospital of Zhejiang University School of Medicine, The First People's Hospital of Yuhang District.
Neuroreport. 2021 Dec 8;32(17):1395-1402. doi: 10.1097/WNR.0000000000001730.
Hypoxic/ischemic brain damage (HIBD) results in increased neonatal mortality and serious neurologic morbidity. Long noncoding RNAs (lncRNAs) are shown as essential modulators of various neurological diseases. Here, we determined the mechanisms of lncRNA GAS5 in mitochondrial apoptosis in HIBD rats.
The HIBD neonatal rat model was established and treated with shRNA-GAS5 or antagomir miR-128-3p. The morphological changes and apoptosis rate were observed by histological staining. Expressions of GAS5, miR-128-3p, and Bax mRNA in brain tissues of HIBD neonatal rats were determined. The binding relationships between GAS5 and miR-128-3p, and miR-128-3p and Bax were confirmed by dual-luciferase assay. Subsequently, the mitochondrial membrane potential and apoptosis-related factors in brain tissues of HIBD neonatal rats were detected. Western blot analysis was performed to detect the expression of Akt/GSK3β pathway-associated proteins.
The neurons in the brain tissue of HIBD neonatal rats decreased with disordered arrangement, and showed vacuolization and nuclear pyknosis, obvious brain damage, increased neuronal apoptosis, and enhanced mitochondrial apoptotic pathway. Downregulated miR-128-3p and upregulated GAS5 and Bax mRNA were found in HIBD neonatal rats. There were binding relationships between GAS5 and miR-128-3p, and miR-128-3p and Bax mRNA. Inhibition of lncRNA GAS5 in HIBD neonatal rats suppressed mitochondrial apoptosis. miR-128-3p knockdown annulled the inhibitory effect of inhibiting lncRNA GAS5 on mitochondrial apoptosis. Silencing GAS5 increased the phosphorylation levels of Akt and GSK3β.
Downregulation of lncRNA GAS5 prevents mitochondrial apoptosis in neonatal HIBD rats by regulating the miR-128-3p/Bax/Akt/GSK-3β axis.
缺氧缺血性脑损伤（HIBD）导致新生儿死亡率增加和严重的神经系统发病率。长链非编码 RNA（lncRNA）被证明是各种神经疾病的重要调节因子。在这里，我们确定了 lncRNA GAS5 在 HIBD 大鼠线粒体凋亡中的作用机制。
建立 HIBD 新生大鼠模型，并给予 shRNA-GAS5 或 antagomir miR-128-3p 处理。通过组织学染色观察形态变化和细胞凋亡率。检测 HIBD 新生大鼠脑组织中 GAS5、miR-128-3p 和 Bax mRNA 的表达。通过双荧光素酶报告实验证实 GAS5 与 miR-128-3p 以及 miR-128-3p 与 Bax 之间的结合关系。随后，检测 HIBD 新生大鼠脑组织中线粒体膜电位和凋亡相关因子。Western blot 分析检测 Akt/GSK3β 通路相关蛋白的表达。
HIBD 新生大鼠脑组织中的神经元排列紊乱，出现空泡化和核固缩，脑损伤明显，神经元凋亡增加，线粒体凋亡途径增强。下调的 miR-128-3p 和上调的 GAS5 和 Bax mRNA 在 HIBD 新生大鼠中发现。GAS5 与 miR-128-3p 以及 miR-128-3p 与 Bax mRNA 之间存在结合关系。抑制 HIBD 新生大鼠中的 lncRNA GAS5 可抑制线粒体凋亡。抑制 lncRNA GAS5 对线粒体凋亡的抑制作用可被 miR-128-3p 敲低所消除。沉默 GAS5 增加了 Akt 和 GSK3β 的磷酸化水平。
下调 lncRNA GAS5 通过调节 miR-128-3p/Bax/Akt/GSK-3β 轴防止 HIBD 新生大鼠的线粒体凋亡。
