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Circ_0004104 participates in the regulation of ox-LDL-induced endothelial cells injury via miR-942-5p/ROCK2 axis
Yuanyuan Zhang1, Shaojun Wang1, Sicong Guo1
1Cardiovascular Internal Medicine Department, Shengjing Hospital of China Medical University, No. 36, Sanhao Street, Heping District, Shenyang, 110000, China.
Insights
Circular RNA circ_0004104 exacerbates atherosclerosis by promoting human umbilical vein endothelial cell injury via the miR-942-5p/ROCK2 axis. Downregulating circ_0004104 protects against oxidized low-density lipoprotein-induced damage.
Area of Science:
- Molecular Biology
- Cardiovascular Research
- Cell Biology
Background:
- Cardiovascular disease is a leading cause of death in the elderly.
- Atherosclerosis is a major contributor to cardiovascular disease.
- Circular RNA circ_0004104 plays a role in atherosclerosis regulation.
Purpose of the Study:
- To investigate the role of circ_0004104 in oxidized low-density lipoprotein (ox-LDL)-induced human umbilical vein endothelial cell (HUVEC) injury.
- To elucidate the underlying molecular mechanism involving miR-942-5p and Rho associated coiled-coil containing protein kinase 2 (ROCK2).
Main Methods:
- Quantitative real-time polymerase chain reaction (qRT-PCR) for gene expression analysis.
- Cell proliferation (MTT assay), apoptosis (flow cytometry), and angiogenesis (tube formation assay) assays.
- Western blot, ELISA, dual-luciferase reporter assay, RNA immunoprecipitation (RIP), and RNA pull-down assays to determine molecular interactions and protein levels.
Main Results:
- Oxidized low-density lipoprotein (ox-LDL) inhibited HUVEC proliferation and promoted apoptosis.
- Circ_0004104 expression was upregulated in atherosclerosis patients' serum and ox-LDL-treated HUVECs.
- Circ_0004104 downregulation protected HUVECs from ox-LDL-induced injury by modulating miR-942-5p and ROCK2 expression.
Conclusions:
- Circ_0004104 downregulation mitigates ox-LDL-induced HUVEC injury.
- The circ_0004104/miR-942-5p/ROCK2 pathway is a key mechanism in ox-LDL-induced endothelial cell dysfunction.
- Targeting circ_0004104 may offer a therapeutic strategy for atherosclerosis.
Background:
Cardiovascular disease was the most common disease among the elderly with high morbidity and mortality. Circ_0004104 was demonstrated to be involved in the regulation of atherosclerosis.
Methods:
Quantitative real-time polymerase chain reaction was employed to measure the expression of circ_0004104, miR-942-5p and Rho associated coiled-coil containing protein kinase 2 (ROCK2). Cell proliferation was tested by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Cell apoptosis was measured by flow cytometry, and tube formation assay was used to detect the angiogenesis ability of cells. Western blot assay was performed to assess protein levels. Enzyme‑linked immunosorbent assay was used to detect the release of IL-1β and TNF-α. The relationship between miR-942-5p and circ_0004104 or ROCK2 was identified by dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, and RNA pull-down assay.
Results:
Oxidized low-density lipoprotein (ox-LDL) inhibited the proliferation of human umbilical vein endothelial cells (HUVECs) and promoted apoptosis in a dose-dependent manner. Circ_0004104 was increased in serum of atherosclerosis patients and ox-LDL-treated HUVECs, and silence of circ_0004104 promoted the proliferation of ox-LDL-exposed HUVECs and inhibited cell apoptosis. MiR-942-5p downregulation reversed si-circ_0004104-mediated influences in HUVECs upon ox-LDL exposure. ROCK2 was the target of miR-942-5p and circ_0004104 regulated the expression of ROCK2 through sponging miR-942-5p. ROCK2 abated the influences of miR-942-5p in ox-LDL-stimulated HUVECs. Circ_0004104 was increased in the exosomes derived from ox-LDL-exposed HUVECs, and the expression of circ_0004104 was promoted in HUVECs after stimulation with ox-LDL-treated HUVECs cells-derived exosomes.
Conclusion:
Circ_0004104 downregulation receded ox-LDL-induced injury in HUVECs through miR-942-5p and ROCK2.
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