Abstract
Circular RNAs (circRNAs) have been revealed to be involved in the pathology of acute ischemic stroke (AIS). Herein, we aimed to study the role and mechanism of circNCOA4 in ischemic stroke. The neuron-like cell line SK-N-SH of the experiment group was cultured in oxygen–glucose deprivation (OGD) condition. Cell viability and apoptosis were evaluated by cell counting kit-8 and flow cytometry. The oxidative damage and endoplasmic reticulum stress (ERS) were analyzed by measuring the production of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and ERS-related markers. The binding between miR-338-5p and circNCOA4 or PDE4B (Phosphodiesterase 4B) was confirmed using dual-luciferase reporter and RIP assays. The commercial kit was used for exosome separation. The levels of circNCOA4 and PDE4B were increased, while miR-338-5p expression was decreased by OGD stimulation. OGD stimulation resulted in the apoptosis of neurons and induced oxidative damage and ERS, these effects were attenuated by circNCOA4 knockdown, while reinforced by circNCOA4 overexpression. Mechanistically, circNCOA4 acted as a sponge for miR-338-5p, and PDE4B was a target of miR-338-5p. MiR-338-5p inhibition reversed the neuroprotective effects of circNCOA4 silencing on neurons. Besides, miR-338-5p overexpression could abolish OGD-induced neuron injury, which was reversed by PDE4B upregulation. In addition, circNCOA4 was packaged into exosomes and showed potential diagnostic value for acute ischemic stroke (AIS) patients. CircNCOA4 has potential diagnostic value for AIS patients and promoted OGD-induced neuron injury via the miR-338-5p/PDE4B axis, providing a new insight into the pathology of AIS.
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The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
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WQ designed and supervised the study. DW conducted the experiments and drafted the manuscript. SX collected and analyzed the data. XL contributed to the methodology, operated the software, and edited the manuscript. All authors reviewed the manuscript.
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Fig. S1
CircNCOA4 regulated OGD-cause apoptosis, oxidative damage and ERS in neurons via PDE4B. (A–K) SK-N-SH cells were co-transfected oe-circNCOA4 or oe-circNCOA4 and si-PDE4B. (A) CCK-8 assay for cell viability. (B) Flow cytometry for cell apoptosis. (C–E) The oxidative stress was analyzed by measuring the contents of ROS, MDA, and SOD. (F–K) Western blotting analysis of the protein levels of PDE4B, GRP78, CHOP, eIF2α, p-eIF2α and Cleaved-caspase3 in cells. (A-K: one-way ANOVA and Tukey's HSD), n = 3 (TIF 1932 KB)
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Wang, D., Xiao, S., Li, X. et al. CircNCOA4 knockdown attenuates OGD-induced neuron injury through miR-338-5p/PDE4B axis. Exp Brain Res 241, 2561–2574 (2023). https://doi.org/10.1007/s00221-023-06702-w
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DOI: https://doi.org/10.1007/s00221-023-06702-w