BACKGROUND: Circulating microRNAs (miRNAs) have been proved to serve as biomarkers for diagnosis and assessment of prognosis of coronary artery disease (CAD). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a widely-used technique to estimate expression levels of circulating miRNAs. Selection of optimal endogenous control (EC) remains critical to obtain reliable qPCR data of miRNAs expression. However, reference controls for normalization of circulating miRNA in CAD are still lacking. The purpose of this study was to identify stably expressed miRNAs to normalize RT-qPCR data derived from plasma in stable CAD. METHODS: We identified 10 stably expressed candidate ECs by combining miRNA microarray screening and literature screening. These 10 candidate ECs were estimated by RT-qPCR and the data were analyzed by NormFinder and BestKeeper algorithm. RESULTS: Two most stable ECs were identified as EC candidates and they were subsequently validated in another larger cohort. The 2 candidates were also validated by normalizing the expression levels of miR-21. In general, they were superior to the commonly used reference gene RNU6 in quantification cycle (Cq) value, stability value and normalization effect. CONCLUSIONS: Our results demonstrated that miR-6090 and miR-4516 can be used as reference genes for plasma miRNA analysis in stable CAD.
Objective: To observe the effect of tablet Yangxintongmai on atherosclerosis( AS) rats. Methods: 72 Wistar rats were divided into four groups, i.e. normal, model, control and therapy group. The AS model of rats was made by feeding with high grease food and AS plague was observed in the aorta after one month of feeding. Tablet Yangxintongmai was used in the group of therapy; tablet Gypenosides was used in the group of control. The level of serum lipid, blood sugar and insulin were tested after one month; the change of aorta histology was observed by HE staining; the ultrastructure of aorta tissue was observed by transmission electronic microscope; the apoptosis of aorta vascular smooth muscle (VSMC) and the expression of actin were evaluated by immunocytochemistry; the expression of VSMC genes such as c-myc, c-fos, PDGF-A, P53, bcl-2 mRNA were tested by RT-PCR. Results: In the therapy group, the level of TC, TG, plasmviscosity, erythrocyte hematocrit and platelet adhesion ratio(P < 0.05)decreased significantly; the level of HDL (P< 0.01), insulin sensitivity index increased; the plague regressed; the histology of aorto didn't change much; the expression of actin decreased; the gene expression of c-myc, PDGF-A, bcl-2(P < 0.05) were downregulated and P53(P < 0.05) was upregulated. Compared with the model group, the expression of c-fos mRNA was downregulated in the therapy group(P < 0.05). Conclusion: Tablet Yangxintongmai has significant effect on AS and the therapeutic molecular mechanism is inhibition of apoptosis and adjustment of the correlated gene expression.