Research Paper Volume 12, Issue 8 pp 6570—6585

Long non-coding RNA MALAT1 exacerbates acute respiratory distress syndrome by upregulating ICAM-1 expression via microRNA-150-5p downregulation

Figure 2. Overexpression of miR-150-5p downregulates MALAT1 expression to suppress apoptosis of HPMECs and decrease the expression of pro-inflammatory cytokines and adhesion factors. (A) The binding site between MALAT1 and miR-150-5p predicted by the starBase website; (B) The expression of miR-150-5p in peripheral blood samples of healthy controls (n = 46) and patients with ARDS (n = 46), as determined by RT-qPCR. * p < 0.05 vs. peripheral blood samples of healthy controls. (C) Expression of miR-150-5p in normal and LPS-treated HPMECs determined by RT-qPCR. * p < 0.05 vs. the control cells. (D) Luciferase activity of MALAT1-WT and MALAT1-MUT. * p < 0.05 vs. cells transfected with NC. (E) The binding of MALAT1 and miR-150-5p and the binding of MALAT1 and AGO2, as analyzed using RIP assay. * p < 0.05 vs. IgG. (F) Expression of miR-150-5p in HPMECs transfected with miR-150-5p mimic or miR-150-5p mimic + oe-MALAT1 determined by RT-qPCR. (G) Expression of pro-inflammatory cytokines (IL-6, IL-1β and TNF-α) in HPMECs transfected with miR-150-5p mimic or miR-150-5p mimic + oe-MALAT1 determined by ELISA. (H) Expression of endothelial cell adhesion molecules (E-selectin and ICAM-1) in HPMECs transfected with miR-150-5p mimic or miR-150-5p mimic + oe-MALAT1 detected by immunofluorescence (× 200). (I) HPMEC apoptosis upon transfection with miR-150-5p mimic or miR-150-5p mimic + oe-MALAT1 detected by TUNEL assay (× 200). (J) Expression of Bcl-2, Bax, cleaved caspase 3 and Caspase3 in HPMECs transfected with miR-150-5p mimic or miR-150-5p mimic + oe-MALAT1 detected by Western blot analysis. * p < 0.05 vs. cells transfected with miR-150-5p mimic NC or miR-150-5p mimic + oe-MALAT1 NC. The data were measurement data and expressed as mean ± standard deviation. The data between two groups were compared using unpaired t-test and those among multiple groups were analyzed by one-way ANOVA, with Tukey's post hoc test. The cell experiment was repeated three times independently.