Cardiac rehabilitation after AMI and atherogenic miRNAs
Description
Clinical, biochemical, anthropometric parameters, and microRNA data are summarized based on study follow-up time points. Raw data from qPCR is included in a separate Excel sheet.
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miRNA measurement Total RNA was extracted from 200 μL of plasma using the miRNeasy Serum/Plasma Advanced Kit (Qiagen, Hilden, Germany). SYBR green-based real-time quantitative PCR was performed using the QuantStudio 6 Flex Real-Time PCR System (Thermo Scientific, Waltham, MA, USA). Spike-in controls were included in each analysis to ensure the quality of RNA isolation, cDNA synthesis. Interplate calibrators were used for calibration between PCR plate runs. Twelve individual miRCURY LNA miRNA PCR assays (Qiagen, Hilden, Germany) were used for quantitative PCR. As endogenous miRNA controls, hsa-miR-103-3p, hsa-miR-191-5p, and hsa-let-7a-5p were selected. GenEx software (MultiD Analyses AB, Gothenborg, Sweden) was used for miRNA expression analysis. Hsa-miR-202-5p and hsa-miR-23a-5p, with a call rate of less than 50% (more than 50% of the data were invalid for these miRNAs), were excluded from the analysis. Cycle threshold (Ct) values greater than 35, which indicate low miRNA plasma concentrations, were replaced with the value 35. Statistical analysis Relative quantity was performed using values from the end of the first month after AMI (baseline), applying a log transformation to ensure normal distribution. Using crossover analysis, we compared data from the intervention and control phases in both arms. Data were tested for normality using the Shapiro‒Wilk test. The independent variables considered were age, sex, and plasma storage time. Data are presented as least square means ± standard errors. The t-test, or Mann-Whitney test and chi-square were used to compare biochemical parameters. ANCOVA was applied. Linear model with a random effect (patient), and several fixed effects were used. Paired t-test or one sample Wilcoxon was used. The Spearman correlation was used to test the linear correlation between biochemical or anthropometric parameters and miRNA quantity. Multiple linear regression was applied to predict the optimal association of miRNAs. Calculations were performed using SPSS 21 software (Chicago, IL, USA). A P value <0.05 was considered statistically significant.
Institutions
- Institut klinicke a experimentalni mediciny