Neonatology, Institute for Women's Health, University College London, London, UK.
Robinson Research Institute, University of Adelaide, Adelaide, SA, Australia.
Pediatr Res. 2021 Feb;89(3):464-475. doi: 10.1038/s41390-020-0986-3. Epub 2020 Jun 10.
Exposure to inflammation exacerbates injury in neonatal encephalopathy (NE). We hypothesized that brain biomarker mRNA, cytokine mRNA and microRNA differentiate inflammation (E. coli LPS), hypoxia (Hypoxia), and inflammation-sensitized hypoxia (LPS+Hypoxia) in an NE piglet model.
Sixteen piglets were randomized: (i) LPS 2 μg/kg bolus; 1 μg/kg infusion (LPS; n = 5), (ii) Saline with hypoxia (Hypoxia; n = 6), (iii) LPS commencing 4 h pre-hypoxia (LPS+Hypoxia; n = 5). Total RNA was acquired at baseline, 4 h after LPS and 1, 3, 6, 12, 24, 48 h post-insult (animals euthanized at 48 h). Quantitative PCR was performed for cytokines (IL1A, IL6, CXCL8, IL10, TNFA) and brain biomarkers (ENO2, UCHL1, S100B, GFAP, CRP, BDNF, MAPT). MicroRNA was detected using GeneChip (Affymetrix) microarrays. Fold changes from baseline were compared between groups and correlated with cell death (TUNEL) at 48 h.
Within 6 h post-insult, we observed increased IL1A, CXCL8, CCL2 and ENO2 mRNA in LPS+Hypoxia and LPS compared to Hypoxia. IL10 mRNA differentiated all groups. Four microRNAs differentiated LPS+Hypoxia and Hypoxia: hsa-miR-23a, 27a, 31-5p, 193-5p. Cell death correlated with TNFA (R = 0.69; p < 0.01) at 1-3 h and ENO2 (R = -0.69; p = 0.01) at 48 h.
mRNA and miRNA differentiated hypoxia from inflammation-sensitized hypoxia within 6 h in a piglet model. This information may inform human studies to enable triage for tailored neuroprotection in NE.
Early stratification of infants with neonatal encephalopathy is key to providing tailored neuroprotection. IL1A, CXCL8, IL10, CCL2 and NSE mRNA are promising biomarkers of inflammation-sensitized hypoxia. IL10 mRNA levels differentiated all three pathological states; fold changes from baseline was the highest in LPS+Hypoxia animals, followed by LPS and Hypoxia at 6 h. miR-23, -27, -31-5p and -193-5p were significantly upregulated within 6 h of a hypoxia insult. Functional analysis highlighted the diverse roles of miRNA in cellular processes.
暴露于炎症会加重新生儿脑病 (NE) 的损伤。我们假设，在 NE 仔猪模型中，脑生物标志物 mRNA、细胞因子 mRNA 和 microRNA 可区分炎症 (大肠杆菌 LPS)、缺氧 (Hypoxia) 和炎症敏感型缺氧 (LPS+Hypoxia)。
16 头仔猪被随机分为三组：(i) LPS 2μg/kg 推注；1μg/kg 输注（LPS；n=5），(ii) 缺氧时生理盐水（Hypoxia；n=6），(iii) 缺氧前 4 小时开始 LPS（LPS+Hypoxia；n=5）。在基线、LPS 后 4 小时以及损伤后 1、3、6、12、24、48 小时采集总 RNA（48 小时处死动物）。使用定量 PCR 检测细胞因子（IL1A、IL6、CXCL8、IL10、TNFA）和脑生物标志物（ENO2、UCHL1、S100B、GFAP、CRP、BDNF、MAPT）。使用 GeneChip（Affymetrix）微阵列检测 microRNA。将各组之间的基线变化与 48 小时时的细胞死亡（TUNEL）进行比较。
在损伤后 6 小时内，我们观察到 LPS+Hypoxia 和 LPS 组的 IL1A、CXCL8、CCL2 和 ENO2 mRNA 增加，与 Hypoxia 组相比。IL10 mRNA 可区分所有组。有 4 种 microRNA 可区分 LPS+Hypoxia 和 Hypoxia：hsa-miR-23a、27a、31-5p、193-5p。细胞死亡与 TNFA（R=0.69；p<0.01）在 1-3 小时之间相关，与 ENO2（R=-0.69；p=0.01）在 48 小时时相关。
在仔猪模型中，mRNA 和 miRNA 在 6 小时内区分了缺氧与炎症敏感型缺氧。这些信息可能为人类研究提供信息，以实现对新生儿脑病的个体化神经保护。
对新生儿脑病婴儿的早期分层是提供个体化神经保护的关键。IL1A、CXCL8、IL10、CCL2 和 NSE mRNA 是炎症敏感型缺氧的有前途的生物标志物。IL10 mRNA 水平可区分所有三种病理状态；LPS+Hypoxia 动物的基线变化幅度最高，其次是 LPS 和 Hypoxia 在 6 小时时。miR-23、-27、-31-5p 和-193-5p 在缺氧损伤后 6 小时内显著上调。功能分析突出了 miRNA 在细胞过程中的多种作用。
