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MicroRNA Profiling in Coronary Endothelial Cells in Type 2 Diabetic Mice
Author(s) -
Zhang Qian,
TsujiHosokawa Atsumi,
Watanabe Makiko,
Makino Ayako
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.lb516
Subject(s) - microrna , biology , fold change , transfection , real time polymerase chain reaction , gene expression , microbiology and biotechnology , gene , genetics
Coronary endothelial dysfunction is a major cause of cardiac ischemia and it is commonly seen in patients with diabetes. MicroRNAs (miRNAs) are small noncoding RNAs that modulate the expression of multiple mRNAs. There is increasing evidence showing that the change in miRNA expression is implicated in the development and/or progression of cardiovascular diseases. However, the pathological role of miRNAs in coronary endothelial dysfunction in type 2 diabetes (T2D) is still unknown. MicroRNA array was used to profile the expression level of miRNAs in coronary endothelial cells (ECs) isolated from control and T2D mice. Real time PCR analysis was performed to confirm the miRNA array results. Among 491 murine miRNAs tested, 64 up‐regulated and 20 down‐regulated miRNAs were identified and seven miRNAs (miR106b, 210, 221, 222, 342‐5p, 378, 705, fold change>1.25) were selected to confirm the data using real‐time PCR. Real time PCR data revealed that miR378 expression was significantly higher and miR210 and miR342‐5p expression levels were significantly lower in diabetic ECs than in control ECs. To identify the target genes of miR378 and 342‐5p, we transfected miR378 mimic or miRNA342‐5p inhibitor in mouse coronary ECs. We chose 91 genes that are highly expressed in ECs and play crucial roles in endothelial functions, including a) endothelium‐derived relaxing factors and their regulators; b) modifiers of cytosolic, mitochondrial, and endoplasmic Ca 2+ concentration; and c) regulators of EC proliferation/migration/apoptosis. The real‐time PCR data showed that miR378 mimic overexpression altered mRNA levels of 11 genes and miR342‐5p inhibitor in 5 genes. Among those genes, Vegfb, Atp3a3, Stim2 were decreased in coronary ECs transfected with miRNA mimic or inhibitor and in coronary ECs in T2D mice compared to their controls. These data suggest that miR378 and/or miR342‐5p are a potential target to restore coronary endothelial dysfunction in diabetes. Support or Funding Information This work was supported by a grant from the National Heart, Lung, and Blood Institute of the National Institutes of Health (HL142214 to A. Makino). This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .