Knockdown of lncRNA ENST00000609755.1 Confers Protection Against Early oxLDL-Induced Coronary Heart Disease

Figures

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  Figure 1. Differentially expressed lncRNAs in CHD vs. non-CHD subjects. (A) Heat map analysis from 10 samples in the CHD subjects (A1, A2, A3, A4, and A5) and non-CHD subjects (B1, B2, B3, B4, and B5) revealed different lncRNA expression profiles. The red color indicates relatively up-regulated lncRNAs, and the green color represents relatively down-regulated lncRNAs; the black portions indicate no significant difference. (B,C) Olcano plot and scatter plot analysis of differentially expressed lncRNAs in CHD and non-CHD subjects. Red or green points represent up-regulated or down-regulated lncRNAs (fold change >2.0, p < 0.05), respectively.

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  Figure 2. Comparison of the expression levels of lncRNAs between the CHD and non-CHD subjects. Five differentially expressed lncRNAs were validated by qRT-PCR. Data are expressed as means ± SD, (n = 30–100/group), *p < 0.05. Student t-test was used for all comparisons.

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  Figure 3. The expression levels of differentially expressed lncRNAs in HCAECs. (A) Purification identification of HCAECs by vWF. vWF factor (red) and cellular localization (blue) double staining ( ×200 times) of HCAECs. (B) Cell viabilities were compared after culturing at different concentrations of oxLDL for 24, 48, and 72 h (n = 3/group). (C) Comparison of the expression levels of differentially expressed lncRNAs after cell culture at different concentrations of oxLDL for 24, 48, and 72 h (n = 3/group). Values are expressed as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 vs. normal HCAECs; #p < 0.05, ##p < 0.01, ###p < 0.001 vs. 50 μg/ml oxLDL; &&p < 0.01, &&&p < 0.001 vs. 100 μg/ml oxLDL in panels b and c. Data are expressed as means ± SD. one-way ANOVA was used for all comparisons.

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  Figure 4. The function of knockdown of lnc-MICALL2-2 in oxLDL-induced HCAECs injury. (A) Lnc-MICALL2-2 interference fragment screening. *p < 0.05, **p < 0.01, ***p < 0.001 vs. normal HCAECs; ##p < 0.01 vs. HCAECs treated with siRNA-1; &p < 0.05 vs. HCAECs treated with siRNA-2 (n = 3/group). (B) Relative expression of lnc-MICALL2-2 in normal HCAECs, HCAECs treated with siRNA, oxLDL, and oxLDL + siRNA. (C) HCAECs morphology of different treatment groups under electron microscopic observation (n = 3/group). (D) LDH activity in HCAECs culture medium of different treatment groups (n = 3/group). (E) HCAECs cell proliferation of different treatment groups activity by Cell Counting Kit-8 detection (n = 9/group). (F,G) HCAECs apoptosis of different treatment groups by flow cytometry (n = 3/group). Values are expressed as the mean ± SD, *p < 0.05, **p < 0.01, ***p < 0.001 in (B,D,E,G). One-way ANOVA was used for all comparisons.