Department of Newborn Infants, Children's Hospital of Nanjing Medical University, Nanjing, 210008, Jiangsu, China.
Mol Brain. 2020 Oct 2;13(1):133. doi: 10.1186/s13041-020-00671-9.
Hypoxic-ischemic brain injury (HIBD) causes neonatal death and serious neurological disability; however, there are currently no promising therapies for it excepting cooling. Therefore, in this study, we used peptidome analysis to identify differentially expressed peptides in cerebrospinal fluid (CSF) of neonates with HIBD or controls, which may give a foundation for finding new promising drugs of neonatal HIBD. CSF samples were collected from neonates with HIBD (n = 4) or controls (n = 4). ITRAQ LC-MS/MS was used to identify differentially expressed peptides between two groups. A total of 35 differentially expressed peptides from 25 precursor proteins were identified. The 2671.5 Da peptide (HSQFIGYPITLFVEKER), one of the down-regulated peptides in neonatal HIBD, is a fragment of heat shock protein 90-alpha (HSP90α/HSP90AA1). Results of bioinformatics analysis showed that HSP90α/HSP90AA1 was located in the protein-protein interaction (PPI) network hub and was involved in the NOD-LIKE receptor (NLR) signaling pathway. This peptide, we named it Hypoxic-Ischemic Brain Damage Associated Peptide (HIBDAP), is a hydrophilic peptide with high stability and has a long half-life of 3.5 h in mammalian reticulocytes. It was demonstrated that TAT-HIBDAP could successfully enter PC12 cells and further into the nucleus. After HIBDAP pretreatment and 6 h of OGD treatment, low concentrations of HIBDAP increased the survival rate of cells, except 40 μM had a toxic effect. Safe concentrations of HIBDAP reduced pyroptosis of PC12 cells under OGD, except 20 μM had no effect, by suppressing expressions of NLRP3, ASC and Caspase-1 except NLRP1. The results of our study identified the characterization and expression profiles of peptides in CSF of neonatal HIBD. Several meaningful peptides such as HIBDAP may play significant roles in neonatal HIBD and provide new therapeutic targets for neonatal HIBD.
缺氧缺血性脑损伤（HIBD）导致新生儿死亡和严重的神经功能障碍；然而，除了降温之外，目前还没有针对它的有希望的治疗方法。因此，在这项研究中，我们使用肽组学分析来鉴定 HIBD 新生儿或对照组的脑脊液（CSF）中差异表达的肽，这可能为寻找新的有希望的新生儿 HIBD 药物提供基础。收集 HIBD 新生儿（n=4）或对照组（n=4）的 CSF 样本。使用 ITRAQ LC-MS/MS 鉴定两组之间差异表达的肽。从 25 种前体蛋白中鉴定出 35 种差异表达的肽。2671.5 Da 肽（HSQFIGYPITLFVEKER）是新生儿 HIBD 中下调的肽之一，是热休克蛋白 90-α（HSP90α/HSP90AA1）的片段。生物信息学分析结果表明，HSP90α/HSP90AA1 位于蛋白质-蛋白质相互作用（PPI）网络枢纽中，参与 NOD-LIKE 受体（NLR）信号通路。这种肽，我们称之为缺氧缺血性脑损伤相关肽（HIBDAP），是一种亲水性肽，稳定性高，在哺乳动物网织红细胞中的半衰期为 3.5 小时。结果表明，TAT-HIBDAP 可以成功进入 PC12 细胞并进一步进入细胞核。在 HIBDAP 预处理和 6 小时 OGD 处理后，低浓度的 HIBDAP 除 40μM 具有毒性作用外，还增加了细胞的存活率。HIBDAP 的安全浓度可抑制 NLRP3、ASC 和 Caspase-1 的表达，除 NLRP1 外，减少 OGD 下 PC12 细胞的细胞焦亡。我们的研究结果确定了 HIBD 新生儿 CSF 中肽的特征和表达谱。一些有意义的肽，如 HIBDAP，可能在新生儿 HIBD 中发挥重要作用，并为新生儿 HIBD 提供新的治疗靶点。
