Research On The MicroRNA Expression Profile Of Atherosclerosis

ObjectiveTo screen the differentially expressed miRNA between atherosclerosispatients and normal controls.To lay the foundation for understanding the role ofmiRNA in the pathogenesis of atherosclerosis and to further study the interactionbetween miRNA and sPLA2-Ⅱ A.Methods1、 Three inpatients or outpatients with coronary atherosclerosis and threehealthy controls were respectively recruited at Department of Cardiology ofthe Affiliated First Hospital of Xiamen University. Peripheral blood of allcases was collected within48hours after the onset of chest pain or othersymptoms, and then the serum was separated.2、 Total RNA was extracted and qualified RNA was markered bye Hy3TM orHy5TM fluorescence group (miRCURYTM Array Power，Exiqon company).The expressed miRNA was screened by microRNA microarray.3、 To further validate the reliability of microRNA microarray results, weselected eleven differentially expressed miRNAs,two-fold increase(foldchange>2) of the miRNAs were7,ranked as hsa-miR-3655, hsa-miR-548ac,hsa-miR-96-5p, hsa-miR-1, hsa-miR-107, hsa-miR-15b-3p, hsa-miR-126-3p,and two-fold decrease (fold change>2) of the miRNAs were4, ranked ashsa-miR-4682, hsa-miR-610, hsa-miR-504-5p, hsa-miR-186-3p. Real-timequantitative PCR was used to validate using hsa-miR-423-5p as an internalreference, the data obtained using the2-ΔΔCTmethod for analysis. Results1、All of the samples were analyzed by microRNA microarray．232differentiallyexpressed miRNAs were obtained,which included134up-regulated and98down-regulated miRNAs(fold change>2). Five-fold increase（fold change>5）of the miRNAs were15,ten-fold increase（fold change>10）of the miRNAswere1,ranked as hsa-miR-548ac. Five-fold decrease（fold change>5）of themiRNAs were20,ten-fold decrease（fold change>10）of the miRNAs were3,ranked as hsa-miR-3155a、 hsa-miR-3158-3p、 hsa-miR-371b-5p.Overall,there were significant differences in the expression of microRNAbetween experimental group and control group.2、Hsa-miR-3655,hsa-miR-548ac,hsa-miR-96-5p,hsa-miR-1,hsa-miR-107,hsa-miR-15b-3p，hsa-miR-126-3p，hsa-miR-4682，hsa-miR-610，hsa-miR-504-5p，hsa-miR-186-3p were selected for validation by real time-PCR.It wasconfirmed that results of real time-PCR were consistent with the gene chips．Conclusion1、There were significant differences in the expression of miRNA betweenatherosclerosis and normal controls. It has established a complete microRNAexpression profile of atherosclerosis. It provides effective and accuratemolecular basis for exploring the role of miRNA in the pathogenesis ofatherosclerosis.2、The consistency between microRNA microarray analysis and real-time PCRresults may indicate miRNA microarray is true and reliable.3、Lay the foundation for futher researching on miRNA targeting regulatorysPLA2-ⅡA role in the pathogenesis of atherosclerosis.