Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon 34141, Korea.
Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul 03080, Korea.
Department of Internal Medicine, Seoul National University Hospital, Seoul 03080, Korea.
Department of Internal Medicine, Seoul National University College of Medicine; Seoul 03080, Korea.
Department of Internal Medicine, Seoul Metropolitan Government Seoul National University Boramae Medical Center, Seoul 07061, Korea.
Abstract
We investigated the diagnostic value and pathophysiological role of circulating microRNA (miR) in vasospastic angina (VA). We enrolled patients who underwent coronary angiography for chest pain to explore the miR's diagnostic utility. In addition, we investigated the role of miRs in regulating endothelial nitric oxide synthase (eNOS) expression in human coronary artery endothelial cells (hCAECs). Among the 121 patients, 46 were diagnosed with VA (VA group), 26 with insignificant coronary lesions (ICL group), and 49 with atherothrombotic angina (AA group). The VA group showed a significantly higher expression of miR-17-5p, miR-92a-3p, and miR-126-3p than the ICL group. In contrast, miR-221-3p and miR-222-3p were upregulated in the AA group compared to the VA group, and all levels of miR-17-5p, miR-92a-3p, miR-126-3p, miR-145-5p, miR-221-3p, and miR-222-3p differed between the AA group and the ICL group. In the hCAECs, transfection with mimics (pre-miR) of miR-17-5p, miR-92a-3p, and miR-126-3p was associated with eNOS suppression. Additionally, transfection with inhibitors (anti-miR) of miR-92a-3p significantly rescued the eNOS suppression induced by lipopolysaccharide. In conclusion, the circulating miRs not only proved to have diagnostic utility, but also contributed to pathogenesis by eNOS regulation.
Keywords
diagnosismicroRNApathogenesisvasospastic angina
Figures
Figure 1Patient enrolment flow. AA, atherothrombotic angina; CAG, coronary angiography; ICL, insignificant coronary lesion; VA, vasospastic angina.Figure 2Circulating miR profiles according to coronary artery disease category. The miR expression relative to mean of the control miR, c-elegans-39 (miR-39) in the VA group (n = 46), AA group (n = 49), and ICL group (n = 26) is demonstrated. Y-axis scale is log-transformed. AA, atherothrombotic angina; Ct, cycle threshold; ICL, insignificant coronary lesion; miR, microRNA; VA, vasospastic angina. One-way analysis of variance was applied and data are presented as mean ± standard deviation. * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001.Figure 3Distinction of patients according to miR profile. In the receiver operating characteristics curve analysis, the area under the curve and 95% confidence intervals were calculated between (A) VA group (n = 46) group and ICL groups (n = 26), (B) VA group and AA group (n = 49), and (C) AA group and ICL group. AA, atherothrombotic angina; AUC, area under the curve; CI, confidence interval; ICL, Insignificant coronary lesion; VA, vasospastic angina; miR, microRNA.Figure 4The role of miRs in endothelial eNOS protein expression. Densitometry of eNOS after each pre-miR transfection (n = 3) (A), densitometry of KLF2 after each pre-miR transfection (n = 3) (B), and densitometry of eNOS and KLF2 after pre-miR-17-5p and pre-miR-92a-3p co-transfection (n = 3) (C,D). One-way analysis of variance was applied and data are presented as mean ± standard deviation. * indicates p < 0.05, ** indicates p < 0.01, *** indicates p < 0.001. eNOS: Endothelial nitric oxide synthase; miR, microRNA.Figure 5Role of miR modulation in eNOS protein expression in hCAECs treated with the vasculotoxic stimuli, LPS. (A) Relative concentration of miR-17-5p, miR-92a-3p, miR-126-3p, miR-145-5p, miR-221-3p, and miR-222-3p in hCAECs according to LPS concentration (n = 3). (B) Densitometry of eNOS after transfection of each anti-miR-17-5p, anti-miR-92a-3p, anti-miR-126-3p, anti-miR-145-5p, anti-miR-221-3p, and anti-miR-22-3p (n = 4). One-way analysis of variance was applied and data are presented as mean ± standard deviation. * Indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001 compared to the control group with LPS treatment. eNOS, endothelial nitric oxide synthase; hCAECs, human coronary artery endothelial cell; LPS, lipopolysaccharide; miR, microRNA; ns, not significant
그림 출처: PubMed Central 공개 논문 (PMC7290712)
Conflict of interest statement
We declare that all authors have no competing interest. The funding source has no involvement in the current study.
