The difference of microRNA profiles among control, normal high density lipoprotein-treated and proflammatory high density lipoprotein-treated endothelial cells

The potential mechanism were successfully illustrated through detecting different expression of microRNAs in between proinflammatory high density lipprotein (pHDL), nomal high density lipprotein (nHDL) and control group, to study whether microRNAs play important roles in the endothelial dysfunction caused by pHDL. 1.To collect fasting blood of 6 coronary heart disease(CHD) patients with hypercholesterolemia and 6 normal subjects in 1st Affilated Hospital of SYSU. 2.Isolated HDL by density gradient centrifugation and measured its pro-inflammatory state. 3.Used PBS, HDL from normal subjects and CHD patients with hypercholesterolemia to treat HUVECs, and then extracted RNA. 4.Applied microarray method to observe the expression of microRNAs in HUVECs and qRT-PCR validation. 5.All experiments were repeated, with results expressed as mean ± standard deviation. Used one-way ANOVA or Kruskal-Wallis test to analyze variance between groups. Applied GraphPad Prism 5.0 and SPSS 13.0 to analyze experimental data. P<0.05, the difference was statistically significant. We performed miRNA microarray analyses using endothelial-enriched RNAs obtained from the HUVECs after 24-hour nHDL, pHDL or control treatment.we found that 21 (14 upregulated and 7 downregulated) of 3100 miRNAs were significantly altered at 24 hours.