Circ_0004104 knockdown alleviates oxidized low-density lipoprotein-induced dysfunction in vascular endothelial cells through targeting miR-328-3p/TRIM14 axis in atherosclerosis

Abstract

Background Circular RNAs have shown important regulatory roles in cardiovascular diseases, containing atherosclerosis (AS). We intended to explore the role of circ_0004104 in AS using oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cells and its associated mechanism. Methods Real-time quantitative polymerase chain reaction and Western blot assay were conducted to analyze RNA levels and protein levels, respectively. Cell viability, apoptosis, angiogenic ability and inflammatory response were assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, flow cytometry, capillary-like network formation assay and enzyme-linked immunosorbent assay, respectively. Cell oxidative stress was assessed using commercial kits. Dual-luciferase reporter assay, RNA immunoprecipitation assay and RNA-pull down assay were performed to verify the intermolecular interaction. Results ox-LDL exposure up-regulated the level of circ_0004104 in HUVECs. ox-LDL exposure suppressed cell viability and angiogenic ability whereas promoted the apoptosis, inflammation and oxidative stress of HUVECs partly through up-regulating circ_0004104. MicroRNA-328-3p (miR-328-3p) was confirmed as a target of circ_0004104. MiR-328-3p interference largely reversed circ_0004104 silencing-mediated effects in HUVECs upon ox-LDL exposure. MiR-328-3p interacted with the 3′ untranslated region of tripartite motif 14, and circ_0004104 positively regulated TRIM14 expression by sponging miR-328-3p. TRIM14 overexpression largely overturned miR-328-3p accumulation-induced influences in HUVECs upon ox-LDL exposure. Conclusion Circ_0004104 knockdown attenuated ox-LDL-induced dysfunction in HUVECs via miR-328-3p-mediated regulation of TRIM14.

Circ_ROBO2/miR-186-5p/TRIM14 axis regulates oxidized low-density lipoprotein-induced cardiac microvascular endothelial cell injury

“…Additionally, Huang et al reported that TRIM14 could promote the activation of endothelium by regulating NF-κB signaling pathway [ 44 ]. Zhang et al found that TRIM14 overexpression largely overturned the protective role of miR-328-3p in ox-LDL-triggered dysfunction in human umbilical vein endothelial cells [ 21 ]. Here, an upregulation of TRIM14 was observed in CAD patient and ox-LDL-exposed CMECs.…”

“…MiR-186-5p was reported to play a critical role in cardiovascular diseases [ 20 ]. Moreover, a recent study indicated that tripartite motif containing 14 (TRIM14) was involved in the progression of atherosclerosis [ 21 ]. Coincidently, we first found that circ_ROBO2 and TRIM14 shared the complementary binding sites for miR-186-5p using bioinformatics tools, which prompted us to establish a circ_ROBO2/miR-186-5p/TRIM14 regulatory network.…”

Emerging Roles of Extracellular Vesicle-Delivered Circular RNAs in Atherosclerosis

“…In addition, exosomes from platelets, vessel cells ( He et al, 2018 ; Bai et al, 2020b ), inflammatory adipocytes ( Wadey et al, 2019 ) and dendritic cells increased the level of inflammatory factors and recruited inflammatory cells and promoted their adhesion to the vessel wall, leading to a chronic inflammatory response process in AS. Multiple circRNAs (circ_0004104 ( Zhang et al, 2021a ), circular ANRIL ( Song et al, 2017 ), circTM7SF3 ( Wang and Bai, 2021 ), circUSP36 ( Miao et al, 2021 ) and so on) were upregulated in AS and demonstrated to induce vascular EC injury and oxidative stress and inflammation. Circ_GRN and Circ_CHFR were also upregulated in atherosclerotic serum and expedited the inflammation of human vascular muscle cells ( Zhuang et al, 2020 ; Li et al, 2021a ).…”

MicroRNAs in peripheral artery disease: potential biomarkers and pathophysiological mechanisms

“…In fact, knock-down of Circ_0004104 led to an increase in miR-328-3p levels, which further resulted in a reduction of ox-LDL. 95 miR-206 is acted on similarly by CircTM7SF3, another circulating RNA. As CircTM7SF3 levels are reduced, miR-206 levels increase, which results in a reduction of ox-LDL.…”