CircCHMP5 Contributes to Ox-LDL-induced Endothelial Cell Injury Through the Regulation of MiR-532-5p/ROCK2 axis

Cardiovasc Drugs Ther. 2023 Dec;37(6):1-12. doi: 10.1007/s10557-022-07316-0. Epub 2022 Jan 27.

Abstract

Background: Aberrant expression of circular RNA (circRNA) has been demonstrated to be related to atherosclerosis (AS) formation. However, the mechanism of circCHMP5 (also known as hsa_circ_0003575) in AS formation remains unclear.

Methods: Oxidized low-density lipoprotein (ox-LDL) was used to treat human umbilical vein endothelial cells (HUVECs) to construct a cell injury model. The expression level of circCHMP5, miR-532-5p, and Rho-associated protein kinase 2 (ROCK2) was measured using quantitative real-time PCR. Cell cycle, apoptosis, proliferation, and angiogenesis were determined by flow cytometry, 5-ethynyl-2'-deoxyuridine (EdU) assay, and tube formation assay. In addition, the protein expression of apoptosis markers, inflammation factors, and ROCK2 was detected by western blot analysis. The interaction between miR-532-5p and circCHMP5 or ROCK2 was assessed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay.

Results: Our results indicated that circCHMP5 was overexpressed in ox-LDL-induced HUVECs. CircCHMP5 knockdown promoted cell cycle, proliferation, and angiogenesis while inhibiting apoptosis and inflammation in ox-LDL-induced HUVECs. MiR-532-5p could be sponged by circCHMP5, and its inhibitor reversed the negative regulation of si-circCHMP5 on ox-LDL-induced HUVECs injury. ROCK2, a target of miR-532-5p, reversed the inhibition effect of miR-532-5p on ox-LDL-induced HUVECs injury. Furthermore, we confirmed that circCHMP5 upregulated ROCK2 by sponging miR-532-5p.

Conclusion: To sum up, our data showed that circCHMP5 regulated the miR-532-5p/ROCK2 axis to accelerate ox-LDL-induced HUVECs injury, confirming that circCHMP5 might be a potential target for AS treatment.

Keywords: Atherosclerosis; CircCHMP5; MiR-532-5p; Ox-LDL; ROCK2.

MeSH terms

  • Apoptosis
  • Atherosclerosis* / genetics
  • Cell Proliferation
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Inflammation
  • Lipoproteins, LDL / toxicity
  • MicroRNAs* / genetics
  • rho-Associated Kinases

Substances

  • oxidized low density lipoprotein
  • Lipoproteins, LDL
  • MicroRNAs
  • ROCK2 protein, human
  • rho-Associated Kinases
  • MIRN532 microRNA, human