1. Serial blood cytokine and chemokine mRNA and microRNA over 48 h are insult specific in a piglet model of inflammation-sensitized hypoxia-ischaemia.
- Author
-
Lingam I, Avdic-Belltheus A, Meehan C, Martinello K, Ragab S, Peebles D, Barkhuizen M, Tann CJ, Tachtsidis I, Wolfs TGAM, Hagberg H, Klein N, Fleiss B, Gressens P, Golay X, Kramer BW, and Robertson NJ
- Subjects
- Animals, Animals, Newborn, Biomarkers, Brain pathology, Chemokines biosynthesis, Chemokines genetics, Cytokines biosynthesis, Disease Models, Animal, Endotoxemia blood, Endotoxemia chemically induced, Gene Expression Regulation, Gene Ontology, Humans, Hypoxia-Ischemia, Brain pathology, Inflammation genetics, Lipopolysaccharides toxicity, Male, Nerve Tissue Proteins biosynthesis, Nerve Tissue Proteins genetics, Phosphopyruvate Hydratase biosynthesis, Phosphopyruvate Hydratase genetics, Random Allocation, Sepsis-Associated Encephalopathy blood, Sepsis-Associated Encephalopathy chemically induced, Sepsis-Associated Encephalopathy pathology, Swine, Time Factors, Tissue Array Analysis, Tumor Necrosis Factor-alpha biosynthesis, Tumor Necrosis Factor-alpha genetics, Cytokines genetics, Hypoxia-Ischemia, Brain blood, Inflammation blood, MicroRNAs blood, RNA, Messenger blood
- Abstract
Background: 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., Methods: 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., Results: 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., Conclusions: 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., Impact: 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.
- Published
- 2021
- Full Text
- View/download PDF