Circvrk1 downregulation attenuates brain microvascular endothelial cell damage induced by oxygen-glucose deprivation through modulating the miR-150-5p/MLLT1 axis

Exp Brain Res. 2023 Mar;241(3):781-791. doi: 10.1007/s00221-023-06555-3. Epub 2023 Feb 3.

Abstract

The pivotal regulatory role of circular RNAs (circRNAs) in ischemic stroke (IS) has been expounded. The study aimed to probe the exact role and underlying mechanism of circVRK1 in oxygen-glucose deprivation (OGD)-induced human brain microvascular endothelial cells (HBMECs) injury. HBMECs challenged by OGD were used as in vitro models of IS. Quantitative real-time PCR was used to examine the levels of circVRK1, vaccinia-related kinase 1 (VRK1), miR-150-5p and MLLT1 mRNA. Cell viability, migration angiogenesis ability and death were evaluated by Cell counting kit-8 assay, transwell assay, wound-healing assay, tube formation assay and flow cytometry analysis. All the protein levels were monitored by western blot assay. Enzyme-linked immunosorbent assay was conducted for examining cell oxidative stress. Dual-luciferase reporter assay, RIP assay and RNA pull-down assay were performed to verify the combination between miR-150-5p and circVRK1 or MLLT1. CircVRK1 was upregulated in OGD-treated HBMECs. CircVRK1 knockdown alleviated OGD-caused effects on HBMECs migration, angiogenesis, death, inflammatory response and oxidative stress. Furthermore, circVRK1 could sponge miR-150-5p, and miR-150-5p silencing also mitigated the impact of circVRK1 deficiency on OGD-evoked injury. Besides, MLLT1 acted as a molecular target of miR-150-5p, and the protective influence of miR-150-5p on OGD-induced cell damage was overturned by MLLT1 introduction. CircVRK1 knockdown weakened OGD-evoked injury in HBMECs through modulating miR-150-5p/MLLT1 pathway, and this might supply new insights and probable targets for IS treatment.

Keywords: Circvrk1; HBMECs; Ischemic stroke; MLLT1; miR-150-5p.

MeSH terms

  • Brain
  • Brain Injuries*
  • Down-Regulation
  • Endothelial Cells
  • Humans
  • Intracellular Signaling Peptides and Proteins
  • MicroRNAs*
  • Neoplasm Proteins
  • Nuclear Proteins
  • Protein Serine-Threonine Kinases
  • Transcription Factors

Substances

  • MicroRNAs
  • MLLT1 protein, human
  • Neoplasm Proteins
  • Nuclear Proteins
  • Transcription Factors
  • VRK1 protein, human
  • Protein Serine-Threonine Kinases
  • Intracellular Signaling Peptides and Proteins
  • MIRN150 microRNA, human