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MicroRNA-21-5p Reduces Hypoxia/Reoxygenation-Induced Neuronal Cell Damage through Negative Regulation of CPEB3

Wang, Bin, Yu, Ping, Lin, Wei, Zhai, Zhaohui·2021·Analytical Cellular Pathology
DOI 10.1155/2021/5543212PMC PMC8660214
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Abstract

Objectives. To explore the role of microRNA-21-5p (miR-21-5p) in hypoxia/reoxygenation- (H/R-) induced HT22 cell damage. Methods. The hypoxia/reoxygenation (H/R) model was established in mouse neuronal cells HT22. Cell Counting Kit-8 (CCK-8) and qRT-PCR were used to determine the effects of H/R treatment on cell viability and miR-21-5p expression. HT22 cells were transfected with miR-21-5p mimic or negative control (NC) followed by the induction of H/R; cell viability, apoptosis, and SOD, MDA, and LDH activities were detected. Besides, the apoptosis-related proteins including BAX, BCL2, cleaved caspase-3, and caspase-3 as well as proteins of EGFR/PI3K/AKT signaling pathways were measured by Western blot. To verify the target relation between cytoplasmic polyadenylation element binding protein 3 (CPEB3) and miR-21-5p, luciferase reporter gene experiment was performed. After cotransfection with miR-21-5p mimic and CPEB3 plasmids, the reversal effects of CPEB3 on miR-21-5p in H/R damage were studied. Results. H/R treatment could significantly reduce the cell viability ( P < 0.05 ) and miR-21-5p levels ( P < 0.05 ) in HT22 cells. After overexpressing miR-21-5p, cell viability was increased ( P < 0.05 ) under H/R treatment, and the apoptosis rate and the levels of apoptosis-related proteins were suppressed (all P < 0.05 ). Furthermore, SOD activity was increased ( P < 0.05 ), while MDA and LDH activity was decreased (both P < 0.05 ). Besides, miR-21-5p could restore the activation of the EGFR/PI3K/AKT signaling pathway inhibited by H/R treatment (all P < 0.05 ). The luciferase reporter gene experiment verified that CPEB3 is the target of miR-21-5p ( P < 0.05 ). When coexpressing miR-21-5p mimic and CPEB3 in the cells, the protective effects of miR-21-5p under H/R were reversed (all P < 0.05 ), and the activation of the EGFR/PI3K/AKT pathway was also inhibited (all P < 0.05 ). Conclusion. This study showed that miR-21-5p may regulate the EGFR/PI3K/AKT signaling pathway by targeting CPEB3 to reduce H/R-induced cell damage and apoptosis.

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Figure 1 The effect of H/R induction on the viability of HT22 cells and the expression of miR-21-5p (n = 3). (a) Changes in the viability of HT22 cells under H/R conditions. (b) Changes in the expression of miR-21-5p in HT22 cells under H/R conditions. Note: Student's t-test was used to compare the H/R group with the control group, aP < 0.05.
Figure 1 The effect of H/R induction on the viability of HT22 cells and the expression of miR-21-5p (n = 3). (a) Changes in the viability of HT22 cells under H/R conditions. (b) Changes in the expression of miR-21-5p in HT22 cells under H/R conditions. Note: Student's t-test was used to compare the H/R group with the control group, aP < 0.05.CC BY · PubMed Central
Figure 2 The effect of miR-21-5p overexpression on H/R-induced cell damage (n = 3). (a) Verification of the overexpression of miR-21-5p in cells transfected with miR-21-5p mimic. (b) The effects of overexpression of miR-21-5p on cell viability in HT22 cells under H/R conditions. Note: Student's t-test was used. Compared with the control group, aP < 0.05; compared with the H/R group, bP < 0.05.
Figure 2 The effect of miR-21-5p overexpression on H/R-induced cell damage (n = 3). (a) Verification of the overexpression of miR-21-5p in cells transfected with miR-21-5p mimic. (b) The effects of overexpression of miR-21-5p on cell viability in HT22 cells under H/R conditions. Note: Student's t-test was used. Compared with the control group, aP < 0.05; compared with the H/R group, bP < 0.05.CC BY · PubMed Central
Figure 3 The effect of miR-21-5p overexpression on H/R-induced apoptosis (n = 3). (a) Under H/R conditions, the effect of overexpression of miR-21-5p in HT22 cells on cell apoptosis; (b) under H/R conditions, changes in the expression of apoptosis-related proteins in cells overexpressing miR-21-5p. The experimental groups are as follows: 1: control group; 2: H/R group; 3: NC+H/R group; 4: miR-21-5p mimic+H/R group. Note: Student's t-test was used. Compared with the control group, aP < 0.05; compared with the H/R group, bP < 0.05.
Figure 3 The effect of miR-21-5p overexpression on H/R-induced apoptosis (n = 3). (a) Under H/R conditions, the effect of overexpression of miR-21-5p in HT22 cells on cell apoptosis; (b) under H/R conditions, changes in the expression of apoptosis-related proteins in cells overexpressing miR-21-5p. The experimental groups are as follows: 1: control group; 2: H/R group; 3: NC+H/R group; 4: miR-21-5p mimic+H/R group. Note: Student's t-test was used. Compared with the control group, aP < 0.05; compared with the H/R group, bP < 0.05.CC BY · PubMed Central
Figure 4 The effects of miR-21-5p overexpression on the activation of the EGFR/PI3K/AKT signaling pathway (n = 3). The experimental groups are as follows: 1: control group; 2: H/R group; 3: NC+H/R group; 4: miR-21-5p mimic+H/R group. Note: Student's t-test was used. Compared with the control group, aP < 0.05; compared with the H/R group, bP < 0.05.
Figure 4 The effects of miR-21-5p overexpression on the activation of the EGFR/PI3K/AKT signaling pathway (n = 3). The experimental groups are as follows: 1: control group; 2: H/R group; 3: NC+H/R group; 4: miR-21-5p mimic+H/R group. Note: Student's t-test was used. Compared with the control group, aP < 0.05; compared with the H/R group, bP < 0.05.CC BY · PubMed Central
Figure 5 The verification of CPEB3 as a target of miR-21-5p (n = 3). (a) CPEB3 as the target of miR-21-5p. (b) Luciferase reporter gene verification. (c) Protein expression level of CPEB3 when miR-21-5p was overexpressed in HT22 cells. (d) The expression level of CPEB3 mRNA when miR-21-5p was overexpressed in HT22 cells. The experimental groups are as follows: 1: control group; 2: H/R group; 3: NC+H/R group; 4: miR-21-5p mimic+H/R group. Note: Student's t-test was used. Compared with the control group, aP < 0.05; compared with the H/R group, bP < 0.05; compared with the NC group, cP < 0.05.
Figure 5 The verification of CPEB3 as a target of miR-21-5p (n = 3). (a) CPEB3 as the target of miR-21-5p. (b) Luciferase reporter gene verification. (c) Protein expression level of CPEB3 when miR-21-5p was overexpressed in HT22 cells. (d) The expression level of CPEB3 mRNA when miR-21-5p was overexpressed in HT22 cells. The experimental groups are as follows: 1: control group; 2: H/R group; 3: NC+H/R group; 4: miR-21-5p mimic+H/R group. Note: Student's t-test was used. Compared with the control group, aP < 0.05; compared with the H/R group, bP < 0.05; compared with the NC group, cP < 0.05.CC BY · PubMed Central
Figure 6 The effect of simultaneous overexpression of miR-21-5p and CPEB3 on H/R-induced cell damage (n = 3). (a) The effect of simultaneous overexpression of miR-21-5p and CPEB3 on H/R-induced cell viability. (b) The effect of cotransfection of cells with miR-21-5p mimic and CPEB3 on H/R-induced apoptosis. (c) The effect of simultaneous overexpression of miR-21-5p and CPEB3 on the expression of apoptosis-related proteins in H/R-induced cell. The experimental groups are as follows: 1: NC+H/R group; 2: miR-21-5p mimic+H/R group; 3: miR-21-5p mimic+OE-NC+H/R group; 4: miR-21-5p mimic+CPEB3+H/R group. Note: Student's t-test was used. Compared with the NC+H/R group, dP < 0.05; compared with the miR-21-5p mimic+H/R group, eP < 0.05.
Figure 6 The effect of simultaneous overexpression of miR-21-5p and CPEB3 on H/R-induced cell damage (n = 3). (a) The effect of simultaneous overexpression of miR-21-5p and CPEB3 on H/R-induced cell viability. (b) The effect of cotransfection of cells with miR-21-5p mimic and CPEB3 on H/R-induced apoptosis. (c) The effect of simultaneous overexpression of miR-21-5p and CPEB3 on the expression of apoptosis-related proteins in H/R-induced cell. The experimental groups are as follows: 1: NC+H/R group; 2: miR-21-5p mimic+H/R group; 3: miR-21-5p mimic+OE-NC+H/R group; 4: miR-21-5p mimic+CPEB3+H/R group. Note: Student's t-test was used. Compared with the NC+H/R group, dP < 0.05; compared with the miR-21-5p mimic+H/R group, eP < 0.05.CC BY · PubMed Central
Figure 7 The effects of overexpressing miR-21-5p and CPEB3 simultaneously on the activation of the EGFR/PI3K/AKT signaling pathway (n = 3). The experimental groups are as follows: 1: NC+H/R group; 2: miR-21-5p mimic+H/R group; 3: miR-21-5p mimic+OE-NC+H/R group; 4: miR-21-5p mimic+CPEB3+H/R group. Note: Student's t-test was used. Compared with the NC+H/R group, dP < 0.05; compared with the miR-21-5p mimic+H/R group, eP < 0.05.
Figure 7 The effects of overexpressing miR-21-5p and CPEB3 simultaneously on the activation of the EGFR/PI3K/AKT signaling pathway (n = 3). The experimental groups are as follows: 1: NC+H/R group; 2: miR-21-5p mimic+H/R group; 3: miR-21-5p mimic+OE-NC+H/R group; 4: miR-21-5p mimic+CPEB3+H/R group. Note: Student's t-test was used. Compared with the NC+H/R group, dP < 0.05; compared with the miR-21-5p mimic+H/R group, eP < 0.05.CC BY · PubMed Central
Figure 8 Diagram showing the roles of miR-21-5p/CPEB3 in H/R-induced HT22 cell damage.
Figure 8 Diagram showing the roles of miR-21-5p/CPEB3 in H/R-induced HT22 cell damage.CC BY · PubMed Central

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Research works printed but not linked: 26

  1. L. B. Goldstein, 2020, Introduction for focused updates in cerebrovascular disease, Stroke, 51, 708, 10.1161/STROKEAHA.119.024159 · DOI
  2. 10.1161/STROKEAHA.118.022015 · DOI
  3. 10.1007/s10571-015-0202-x · DOI
  4. 10.3390/antiox8100454 · DOI
  5. 10.1016/j.ejphar.2017.04.002 · DOI
  6. L. Yang, 2015, MicroRNA-323 regulates ischemia/reperfusion injury-induced neuronal cell death by targeting BRI3, International Journal of Clinical and Experimental Pathology, 8, 10725
  7. 10.1186/s12974-017-0977-4 · DOI · Paper page
  8. K. Saliminejad, 2019, An overview of microRNAs: biology, functions, therapeutics, and analysis methods, Journal of cellular physiology, 201, 5451, 10.1002/jcp.27486 · DOI · Paper page
  9. 10.1177/0300985813502820 · DOI
  10. 10.1007/978-3-319-22671-2_9 · DOI
  11. 10.1016/j.yexcr.2017.01.015 · DOI
  12. 10.5732/cjc.30.0371 · DOI
  13. 10.1007/s12031-018-1067-5 · DOI
  14. 10.1007/s00210-014-1006-0 · DOI
  15. 10.3390/ijms160922368 · DOI
  16. 10.1016/j.ejphar.2019.01.022 · DOI
  17. 10.1152/physiolgenomics.00020.2014 · DOI
  18. 10.1152/physiolgenomics.00130.2010 · DOI
  19. 10.1007/s13277-016-5035-9 · DOI
  20. E. Candelario-Jalil, 2009, Injury and repair mechanisms in ischemic stroke: considerations for the development of novel neurotherapeutics, Current Opinion in Investigational Drugs, 10, 644
  21. 10.2174/1567202612666150311104900 · DOI
  22. 10.1007/s00232-009-9177-1 · DOI
  23. 10.1080/02652030500058403 · DOI
  24. 10.1002/cbf.2977 · DOI
  25. 10.3892/or.2019.6983 · DOI
  26. 10.1002/jcb.28528 · DOI

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Metadata

Authors
Wang, B., Yu, P., Lin, W., & Zhai, Z.
Affiliations
  • Department of Neurology, Affiliated Hospital of Weifang Medical College, Weifang, Shandong 261031, China
  • Pain Department, Weifang People’s Hospital, Weifang, Shandong 261041, China
  • Department of Plastic Surgery, Plastic Surgery Hospital of Weifang Medical University, Weifang, Shandong 261042, China
ORCID
Venue
Analytical Cellular Pathology
Publisher
Wiley
Published
2021-12-02
Volume
2021
Pages
1-11
ISSN
  • 2210-7185
  • 2210-7177
DOI
10.1155/2021/5543212
PMC
PMC8660214
Cited by
5
References
0
Language
en
Licence
creativecommons.org/licenses/by/4.0/ (unspecified)
Funders
Shandong Province Medicine and Health Technology Development Plan (2018WS062)

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