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Methods: 8-week-old C57BL/6 mice were treated with intratracheal bleomycin or PBS. Lungs were collected at 4 or 7 days. miRNA expression was compared using a qPCR based 56-miRNA array. TargetScan and Enrichr were used to identify predicted gene targets and biological processes of 4 highly down-regulated miRNAs. Expression of 4 predicted miR-26a gene targets, EphA2, VEGFC, KDR and ROCK1 was evaluated by qPCR. EphA2 protein levels were measured by western blot analysis. In situ hybridization was performed to localize lung miR-26a. Human lung microvascular endothelial cells (HMVEC-L) were transfected with miR-26a mimic, inhibitor and negative controls. miR-26a, EphA2 mRNA/protein and cell permeability were measured.

Results: Bleomycin decreased 4 miRNAs (miR-1, miR-26a, miR-30b and miR-30c) by greater than 6 fold. Of these miRNAs, miR-26a is predicted to regulate 135 …