a Department of Children's Healthcare, East Medical District of Linyi People's Hospital , Linyi , China.
b Department of Internal Medicine, Shandong Provincial Coal Linyi Hot Springs Sanatorium Hospital , Linyi , China.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):2492-2499. doi: 10.1080/21691401.2019.1623228.
Neonatal hypoxia-ischemia is a troublesome disease. Angelica polysaccharide (AP) is proved to have antioxidant effects. Our study was performed to confirm the effects of AP in hypoxia-exposed neural stem cells (NSCs). NSCs were pre-treated with AP and then stimulated with hypoxia. Viability of NSCs was examined by Cell Counting Kit-8 assay. Hypoxia-introduced apoptosis was observed by flow cytometry. Essential regulators of mTOR and Notch signalling pathways were examined by Western blot. mRNA expression was accessed using qRT-PCR. Bcl2/adenovirus EIB 19kD-interacting protein 3 (BNIP3) was altered by transfection. We noticed that NSCs were sensitive to hypoxia-induced apoptosis and showed decreased viability. Moreover, Beclin and light chain 3-II was upregulated while p62 was downregulated. However, AP reversed all these results. Similarly, hypoxia decreased the phosphorylation of mTOR and p70S6K and Notch1 expression while AP increased the phosphorylation of mTOR and p70S6K as well as the expression of Notch1. BNIP3 was upregulated by hypoxia while downregulated by AP. Further experiments demonstrated that overexpression of BNIP3 broken all the effects induced by AP shown in cell viability, apoptosis, autophagy and signalling pathways. Collectively, AP alleviated hypoxia-introduced NSCs damages by maintaining cell viability, blocking apoptosis and autophagy via downregulation of BNIP3 with the activation of mTOR and Notch signalling pathways.
新生儿缺氧缺血是一种棘手的疾病。当归多糖（AP）已被证明具有抗氧化作用。我们的研究旨在证实 AP 在缺氧暴露的神经干细胞（NSC）中的作用。用 AP 预处理 NSC，然后用缺氧刺激。通过细胞计数试剂盒-8 测定法检查 NSC 的活力。通过流式细胞术观察缺氧诱导的细胞凋亡。通过 Western blot 检查 mTOR 和 Notch 信号通路的基本调节因子。使用 qRT-PCR 检测 mRNA 表达。通过转染改变 Bcl2/腺病毒 EIB 19kD 相互作用蛋白 3（BNIP3）。我们注意到 NSC 对缺氧诱导的细胞凋亡敏感，活力下降。此外，Beclin 和 LC3-II 上调，而 p62 下调。然而，AP 逆转了所有这些结果。同样，缺氧降低了 mTOR 和 p70S6K 的磷酸化以及 Notch1 的表达，而 AP 增加了 mTOR 和 p70S6K 的磷酸化以及 Notch1 的表达。BNIP3 由缺氧上调，由 AP 下调。进一步的实验表明，BNIP3 的过表达破坏了 AP 在细胞活力、细胞凋亡、自噬和信号通路中诱导的所有作用。总之，AP 通过下调 BNIP3 并激活 mTOR 和 Notch 信号通路，维持细胞活力、阻断凋亡和自噬，从而减轻缺氧引起的 NSC 损伤。
