Abstract:Aim To identify the correlation between microRNA (miRNA) and low-response to clopidogrel in patients with coronary heart disease (CHD), and explore their diagnostic value as biomarkers of low-response to clopidogrel.Methods 158 patients with stable CHD were recruited as the research population. The maximum platelet aggregation rate (MPAR) induced by adenosine diphosphate (ADP) was measured using light transmission aggregometry for all these patients. Eventually, 35 patients of them whose MPAR≥65% were allocated to low-response group. For comparison, 35 age-and gender-matched patients with MPAR＜65% were allocated to normal-response group. The peripheral blood mononuclear cells (PBMC) were isolated from the blood samples of each patient to extract total RNA. The expression differences of miR-34a-5p, miR-370-3p, miR-432-5p and miR-495-3p in the two groups were detected by real-time quantitative PCR. The diagnostic value of differently expressed miRNA for low-response to clopidogrel were evaluated by correlation analysis and receiver-operating characteristic (ROC) curve analysis. Results Compared with normal-response group, the expressions of miR-34a-5p and miR-495-3p were significantly elevated in low-response group, and their fold changes were greater than 2 (P＜0.05). There was no significant difference in miR-370-3p and miR-432-5p expressions between two groups (P＞0.05). The expressions of miR-34a-5p and miR-495-3p were positively correlated with MPAR in CHD patients (r＝0.709, P＜0.01; r＝0.597, P＜0.01). ROC curve analysis showed that the area under the curve(AUC) of miR-34a-5p was 0.789, and the AUC of miR-495-3p was 0.787; while the AUC of miR-34a-5p+miR-495-3p was 0.873, which was significantly higher than that of miR-34a-5p (P＜0.05) or miR-495-3p (P＜0.05). Conclusion Both the high expression of miR-34a-5p and miR-495-3p were associated with low-response to clopidogrel in CHD patients, and may serve as novel biomarkers for low-response to clopidogrel in CHD.

Abstract:Aim To investigate the correlation between plasma miR-1228-5p, miR-34a-5p, miR-192-5p and miR-30a-3p levels and premature coronary artery disease (PCAD) and their early screening value as biomarkers. Methods According to inclusion criteria and exclusion criteria, 6 patients with clearly diagnosed PCAD were included as PCAD group and 6 heathy subjects as control group. Blood was collected from the PCAD group and the control group, and serum samples were extracted and stored. The miRNA levels in the serum of the two groups were detected using DNBseq platform, and the miRNA with significant differences were screened. According to inclusion and exclusion criteria, blood from 78 patients with PCAD, 75 patients with late-onset coronary artery disease and 69 healthy subjects were collected and the selected miRNAs were verified by qPCR. Coronary angiography reports of patients with PCAD were analyzed, and the severity of coronary artery disease was evaluated by Gensini score. Spearman correlation test was used to analyze the correlation between the degree of coronary stenosis and the miRNA. ROC curve was used to analyze the diagnostic value of plasma levels of miR-1228-5p, miR-34a-5p, miR-192-5p and miR-30a-3p for PCAD, and multivariate Logistic regression was used to analyze the influencing factors of PCAD. Results DNBseq platform analysis showed 33 differentially expressed miRNAs, including 17 up-regulated miRNAs and 16 down-regulated miRNAs. The 5 miRNAs with the most significant difference levels were miR-1228-5p, miR-34a-5p, miR-192-5p, miR-424-3p and miR-30a-3p. qPCR results showed that compared with the control group, plasma miR-1228-5p, miR-34a-5p, miR-192-5p and miR-30a-3p levels were increased by 1.7 times, 1.4 times, 0.7 times and 2.5 times in patients with PCAD (P＜0.05). There was no significant difference in miR-424-3p level between the two groups (P＞0.05). Spearman correlation analysis showed that plasma levels of miR-1228-5p and miR-34a-5p were positively correlated with the degree of coronary stenosis in patients with PCAD (r＝0.307, P＝0.004; r＝0.238, P＝0.036). ROC curve analysis showed that the areas under ROC curve for the diagnosis of PCAD by miR-1228-5p, miR-34a-5p, miR-192-5p and miR-30a-3p were 0.3,0.2,0.731 and 0.798, respectively. The area under ROC curve of the combined diagnosis was 0.990, and 95%CI was 0.970～1.000. Conclusions The plasma levels of miR-1228-5p, miR-34a-5p, miR-192-5p and miR-30a-3p in patients with PCAD were significantly increased. The combined detection showed high accuracy in the diagnosis of PCAD, which is expected to be new biomarkers for preliminary screening of PCAD.