Integrated transcriptomic and proteomic profiling reveals the key molecular signatures of brain endothelial reperfusion injury

Abstract

Abstract Reperfusion after ischemic stroke often causes brain microvascular injury and blood-brain barrier disruption; however, the underlying mechanisms are unclear. Here, we performed transcriptomic and proteomic analyses on human cerebral microvascular endothelial cells following oxygen-glucose deprivation (OGD) or OGD plus recovery (OGD/R), to identify molecules and signaling pathways dysregulated by reperfusion. Transcriptomic analysis identified 390 differentially expressed genes (301 upregulated and 89 downregulated) between the OGD/R and OGD groups. Pathway analysis indicated that the tumor necrosis factor (TNF) signaling pathway was the most significantly enriched. Furthermore, these genes were mostly associated with inflammation, including the TNF signaling pathway, TGF-β signaling pathway, cytokine-cytokine receptor interaction, NOD-like receptor signaling pathway, and NF-κB signaling pathway. On the other hand, 201 differentially expressed proteins (96 upregulated and 105 downregulated) were identified by proteomics between the OGD/R and OGD groups and were primarily associated with extracellular matrix destruction and remodeling, impairment of endothelial transport function, and inflammatory responses. Six genes (DUSP1, JUNB, NFKBIA, NR4A1, SERPINE1, and THBS1) were upregulated by OGD/R at both the mRNA and protein levels. The expression of genes related to inflammatory responses and extracellular matrix were further measured in a mouse model of cerebral ischemia/reperfusion in vivo. Overall, our study provides a comprehensive molecular atlas of brain endothelial reperfusion injury and may facilitate the understanding and treatment of reperfusion injury after ischemic stroke.