Research Paper Volume 12, Issue 21 pp 21076—21090

Downregulation of interleukin-6 and C-reactive protein underlies a novel inhibitory role of microRNA-136-5p in acute lower extremity deep vein thrombosis

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Figure 5. The suppressive effect of miR-136-5p elevation on HUVEC apoptosis in vitro is reversed by IL-6 and CRP. (A) The expression levels of miR-136-5p, IL-6, and CRP in normal and HUVECs exposed to CoCl2 determined by RT-qPCR (* p < 0.05 compared with normal HUVECs). (B) The levels of IL-6 and CRP in the culture medium supernatant of normal and HUVECs exposed to CoCl2 determined by ELISA (* p < 0.05 compared with normal HUVECs). (C) Apoptosis rate of HUVECs following the treatment of miR-136-5p agomir or miR-136-5p antagomir measured using flow cytometry (* p < 0.05 compared with HUVECs treated with NC agomir; # p < 0.05 compared with HUVECs treated with NC antagomir). (D) The apoptosis rate of HUVECs after treatment of miR-136-5p agomir + oe-IL-6, oe-CRP, or oe-NC detected by rescue experiment (* p < 0.05 compared with HUVECs treated with miR-136-5p agomir and oe-NC). Measurement data were expressed as mean ± standard deviation. Data from two groups were compared using independent sample t-test and data from multiple groups using one-way ANOVA. Each experiment was repeated three times. miR-136-5p, microRNA-136-5p; IL-6, interleukin-6; CRP, C-reactive protein; RT-qPCR, reverse transcription quantitative polymerase chain reaction; ELISA, enzyme linked immunosorbent assay; HUVECs, human umbilical vein endothelial cells; NC, negative control; ANOVA, analysis of variance; CoCl2, cobalt chloride.