Changes in MicroRNA Expression Level of Circulating Platelets Contribute to Platelet Defect After Cardiopulmonary Bypass

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Abstract

Objectives:

Platelet defect mechanisms after cardiopulmonary bypass remain unclear. Our hypothesis microRNA expressions in circulating platelets significantly change between pre and post cardiopulmonary bypass, and consequent messenger RNA and protein expression level alterations cause postcardiopulmonary bypass platelet defect.

Design:

Single-center prospective observational study.

Setting:

Operating room of Kyoto Prefectural University of Medicine.

Patients:

Twenty-five adult patients scheduled for elective cardiac surgeries under cardiopulmonary bypass.

Interventions:

None.

Measurements and Main Results:

In the initial phase, changes in microRNA expression between pre and post cardiopulmonary bypass underwent next generation sequencing analysis (10 patients). Based on the results, we focused on changes in mir-10b and mir-96, which regulate glycoprotein 1b and vesicle-associated membrane protein 8, respectively, and followed them until messenger RNA and protein syntheses (15 patients) using quantitative polymerase chain reaction and Western blotting. Seven microRNAs including mir-10b and mir-96 exhibited significant differences in the initial phase. In the subsequent phase, mir-10b-5p and mir-96-5p overexpressions were confirmed, and glycoprotein 1b and vesicle-associated membrane protein 8 messenger RNA levels were significantly decreased after cardiopulmonary bypass: fold differences (95% CI): mir-10b-5p: 1.35 (1.05–2.85), p value equals to 0.01; mir-96-5p: 1.59 (1.06–2.13), p value equals to 0.03; glycoprotein 1b messenger RNA: 0.46 (0.32–0.60), p value of less than 0.001; and vesicle-associated membrane protein messenger RNA: 0.70 (0.56–0.84), p value of less than 0.001. Glycoprotein 1b and vesicle-associated membrane protein 8 were also significantly decreased after cardiopulmonary bypass: glycoprotein 1b: 82.6% (71.3–93.8%), p value equals to 0.005; vesicle-associated membrane protein 8: 79.0% (70.7–82.3%), p value of less than 0.001.

Conclusions:

Expressions of several microRNAs in circulating platelets significantly changed between pre and post cardiopulmonary bypass. Overexpressions of mir-10b and mir-96 decreased glycoprotein 1b and vesicle-associated membrane protein 8 messenger RNA as well as protein, possibly causing platelet defect after cardiopulmonary bypass.

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