Abstract
Ischemic cerebrovascular disease is the main cause of disability due to stroke. This study aimed to investigate the function of miR-1224 in OGD/R-induced hippocampal neuron apoptosis, as well as the regulatory mechanism of miR-1224 in ischemic cerebrovascular disease. The oxygen-glucose deprivation/reperfusion (OGD/R) model of primary mouse hippocampal neurons was established. RT-qPCR detected miR-1224, Ku70 and Ku86 levels. Western blotting was applied to measure the expression of Ku70/86 and apoptosis related proteins. Flow cytometry was used to assess apoptosis. JC-1 fluorescence was performed to test the mitochondrial membrane potential (MMP) in neurons. The double luciferase reporter assay was performed to investigate the relationship between miR-1224 and Ku70/86. OGD/R induced the apoptosis and mitochondrial injury in neuronal cells, while miR-1224 downregulation or Ku70/86 upregulation reversed this phenomenon. Meanwhile, miR-1224 negatively regulated the expression of Ku70/86 in neuronal cells through directly targeting Ku70/86. Furthermore, knockdown of Ku70/86 significantly reversed the inhibitory effect of miR-1224 silencing on apoptosis and mitochondrial injury in OGD/R-treated neuronal cells. Our findings indicated that miR-1224 downregulation suppressed OGD/R-induced hippocampal neuron apoptosis by targeting Ku protein, suggesting that miR-1224 could serve as a new target for ischemic cerebrovascular disease treatment.
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The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.
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Abbreviations
- MiRNA:
-
microRNA
- OGD/R:
-
Oxygen-glucose deprivation/recovery
- I/R:
-
ischemia-reperfusion
- MMP:
-
Mitochondrial membrane potential
- NHEJ:
-
non-homologous end joining
- DNA-PKcs:
-
DNA-dependent protein kinases
- RSF1:
-
remodeling and spacing factor 1
- RT-qPCR:
-
Quantitative real time polymerase chain reaction
- Cyt-c:
-
Cytochrome c
- DMEM:
-
Dulbecco’s modified Eagle medium
- MTT:
-
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide
- DMSO:
-
Dimethyl sulfoxide
- HBSS:
-
Hank’s Balanced Salt Solution
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This work was supported by Hunan Provincial Natural Science Foundation of China (2019JJ50540).
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TX analyzed and interpreted the patient data regarding the hematological disease and the transplant. JW performed the histological examination of the kidney, and JW was a major contributor in writing the manuscript. All authors read and approved the final manuscript.
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All in vivo experiments were performed in accordance with National Institutes of Health guide for the care and use of laboratory animals, following a protocol approved by the Ethics Committees of the First Affiliated Hospital, Hengyang Medical School, University of South China.
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Wan, J., Xiao, T. MiR-1224 downregulation inhibits OGD/R-induced hippocampal neuron apoptosis through targeting Ku protein. Metab Brain Dis 37, 531–543 (2022). https://doi.org/10.1007/s11011-021-00873-7
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DOI: https://doi.org/10.1007/s11011-021-00873-7