The Journal of Practical Medicine >

2023 , Vol. 39 >Issue 23: 3051 - 3057

DOI: https://doi.org/10.3969/j.issn.1006-5725.2023.23.005

Basic Research

Effects of miR⁃27a⁃3p on neuronal apoptosis induced by oxygen glucose deprivation/reoxygenation via targeting Rnd3

  • Junjie LI ,
  • Wenya BAI ,
  • Wendong CHEN ,
  • Wei YANG ,
  • Jianlin. SHAO
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  • Department of Anesthesiology，the First Affiliated Hospital of Kunming Medical University，Kunming 650032，China

Received date: 2023-05-17

  Online published: 2024-01-08

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Abstract

Objective To investigate the effect and mechanism of miR?27a?3p on nerve cell apoptosis induced by oxygen glucose deprivation/reoxygenation （OGD/R） through regulation of Rho family GTPase 3（Rnd3） expression. Methods PC12 neurons were cultured in vitro and reoxygenated for 3， 6， 9 h and 12 h after 2 h oxygen glucose deprivation. Cell viability， miR?27a?3p expression and Rnd3 mRNA expression were assessed at each time point and the optimal reoxygenation time point was screened. After transfection of miR?27a?3p Mimic， miR?27a?3p Inhibitor and their negative control， transfection of shRnd3 and its negative control， or co?transfection of shRnd3 and miR?27a?3p Inhibitor through lentivirus， CCK?8 assay was used to detect cell activity. The apoptosis rate of the cells was detected using flow cytometry. Expression of miR?27a?3p and Rnd3 mRNA was detected by RT?qPCR. Expression of apoptosis?related protein and Rnd3 protein was detected by Western blot. The dual luciferase reporter assay confirmed the targeting relationship between miR?27a?3p and Rnd3. Results Upregulation of miR?27a?3p increased cell viability， decreased total cell apoptosis rate， suppressed pro?apoptotic proteins Cleaved Caspase?3 （C?caspase?3） and Bax， and promoted expression of anti?apoptotic protein Bcl?2 （P < 0.05）； The opposite result was found when down?regulating miR?27a?3p. The double luciferase reporter gene assay showed that Rnd3 was the target gene of miR?27a?3p. Down?regulation of Rnd3 increased cell viability， decreased the total rate of apoptosis， suppressed the pro?apoptotic protein C?caspase?3， Bax， and promoted expression of the anti?apoptotic protein Bcl?2 （P < 0.05）. However， miR?27a?3p Inhibitor reversed the protective effect of shRnd3. Conclusion miR?27a?3p alleviates OGD/R?induced damage to PC12 neurons by targeting Rnd3 to inhibit cell apoptosis.

Key words： miR?27a?3p; OGD/R; Rnd3; cell apoptosis

Cite this article

Junjie LI , Wenya BAI , Wendong CHEN , Wei YANG , Jianlin. SHAO . Effects of miR⁃27a⁃3p on neuronal apoptosis induced by oxygen glucose deprivation/reoxygenation via targeting Rnd3[J]. The Journal of Practical Medicine, 2023 , 39(23) : 3051 -3057 . DOI: 10.3969/j.issn.1006-5725.2023.23.005

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