Skip to main content

Advertisement

SpringerLink
Log in

Silencing of the circ-Arhgap5 RNA protects neuronal PC12 cells against injury and depends on the miR-29a-3p/Rock1 axis

  • Original Article
  • Published:
Metabolic Brain Disease Aims and scope Submit manuscript

Abstract

Circular RNAs (circRNAs) are abundantly expressed in human central nervous system. Here, we explored the role of circ_Arhgap5 in cerebral ischemia/reperfusion (I/R)-induced nerve injury in PC12 cells and its associated mechanism. Cell proliferation ability was assessed by 5-Ethynyl-2’-deoxyuridine (Edu) assay and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry (FCM) was applied to assess cell apoptotic rate. Cell death was analyzed by lactate dehydrogenase (LDH) assay. Oxygen–glucose deprivation and reoxygenation (OGD/R) up-regulates the expression of circRNA Rho GTPase activating protein 5 (circ_Arhgap5; mmu_circ_0000377) in PC12 cells. OGD/R exposure inhibited the proliferation and induced the apoptosis of PC12 cells, and the silence of circ_Arhgap5 attenuated OGD/R-induced injury in PC12 cells. miR-29a-3p was identified as a target of circ_Arhgap5 in PC12 cells. Circ_Arhgap5 knockdown-mediated protective effects in OGD/R-induced PC12 cells were reversed by the interference of miR-29a-3p. miR-29a-3p interacted with the 3’ untranslated region (3’UTR) of Rho-associated coiled-coil-containing protein kinase 1 (Rock1), and circ_Arhgap5 can positively regulate Rock1 expression by sponging miR-29a-3p in PC12 cells. miR-29a-3p overexpression protected PC12 cells against OGD/R-induced damage by down-regulating Rock1. In conclusion, circ_Arhgap5 silencing protected PC12 cells from OGD/R-induced injury through mediating miR-29a-3p/Rock1 axis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Data availability

Not applicable.

References

  • An Q, Liu T, Wang MY, Yang YJ, Zhang ZD, Lin ZJ, Yang B (2020) circKRT7-miR-29a-3p-col1a1 axis promotes ovarian cancer cell progression. Onco Targets Ther 13:8963–8976

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Benjamin EJ, Blaha MJ, Chiuve SE, Cushman M, Das SR, Deo R, de Ferranti SD, Floyd J, Fornage M, Gillespie C, Isasi CR, Jiménez MC, Jordan LC, Judd SE, Lackland D, Lichtman JH, Lisabeth L, Liu S, Longenecker CT, Mackey RH, Matsushita K, Mozaffarian D, Mussolino ME, Nasir K, Neumar RW, Palaniappan L, Pandey DK, Thiagarajan RR, Reeves MJ, Ritchey M, Rodriguez CJ, Roth GA, Rosamond WD, Sasson C, Towfighi A, Tsao CW, Turner MB, Virani SS, Voeks JH, Willey JZ, Wilkins JT, Wu JH, Alger HM, Wong SS, Muntner P (2017) Heart disease and stroke statistics-2017 update: a report from the American Heart Association. Circulation 135:e146–e603

    Article  PubMed  PubMed Central  Google Scholar 

  • Chai Z, Gong J, Zheng P, Zheng J (2020) Inhibition of miR-19a-3p decreases cerebral ischemia/reperfusion injury by targeting IGFBP3 in vivo and in vitro. Biol Res 53:17

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen W, Wang H, Feng J, Chen L (2020a) Overexpression of circRNA circUCK2 attenuates cell apoptosis in cerebral ischemia-reperfusion injury via miR-125b-5p/GDF11 Signaling. Mol Ther Nucleic Acids 22:673–683

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen Y, Zhang W, Yan L, Zheng P, Li J (2020b) miR-29a-3p directly targets Smad nuclear interacting protein 1 and inhibits the migration and proliferation of cervical cancer HeLa cells. PeerJ 8:e10148

    Article  PubMed  PubMed Central  Google Scholar 

  • Chugh C (2019) Acute ischemic stroke: management approach. Indian J Crit Care Med 23:S140-s146

    Article  PubMed  PubMed Central  Google Scholar 

  • Csonka C, Sárközy M, Pipicz M, Dux L, Csont T (2016) Modulation of hypercholesterolemia-induced oxidative/nitrative stress in the heart. Oxid Med Cell Longev 2016:3863726

    Article  PubMed  Google Scholar 

  • Fabian MR, Sonenberg N, Filipowicz W (2010) Regulation of mRNA translation and stability by microRNAs. Annu Rev Biochem 79:351–379

    Article  CAS  PubMed  Google Scholar 

  • Fu J, Wu B, Zhong S, Deng W, Lin F (2020) miR-29a-3p suppresses hepatic fibrosis pathogenesis by modulating hepatic stellate cell proliferation via targeting PIK3R3 gene expression. Biochem Biophys Res Commun 529:922–929

    Article  CAS  PubMed  Google Scholar 

  • Gao XZ, Zhang ZX, Han GL (2020) MiR-29a-3p enhances the viability of rat neuronal cells that injured by oxygen-glucose deprivation/reoxygenation treatment through targeting TNFRSF1A and regulating NF-κB signaling pathway. J Stroke Cerebrovasc Dis 29:105210

    Article  PubMed  Google Scholar 

  • Geng C, Wei J, Wu C (2020) Yap-Hippo pathway regulates cerebral hypoxia-reoxygenation injury in neuroblastoma N2a cells via inhibiting ROCK1/F-actin/mitochondrial fission pathways. Acta Neurol Belg 120:879–892

    Article  PubMed  Google Scholar 

  • Hansen TB, Jensen TI, Clausen BH, Bramsen JB, Finsen B, Damgaard CK, Kjems J (2013) Natural RNA circles function as efficient microRNA sponges. Nature 495:384–388

    Article  CAS  PubMed  Google Scholar 

  • He AT, Liu J, Li F, Yang BB (2021) Targeting circular RNAs as a therapeutic approach: current strategies and challenges. Signal Transduct Target Ther 6:185

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jeyaseelan K, Lim KY, Armugam A (2008) MicroRNA expression in the blood and brain of rats subjected to transient focal ischemia by middle cerebral artery occlusion. Stroke 39:959–966

    Article  CAS  PubMed  Google Scholar 

  • Kristensen LS, Andersen MS, Stagsted LVW, Ebbesen KK, Hansen TB, Kjems J (2019) The biogenesis, biology and characterization of circular RNAs. Nat Rev Genet 20:675–691

    Article  CAS  PubMed  Google Scholar 

  • Krol J, Loedige I, Filipowicz W (2010) The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet 11:597–610

    Article  CAS  PubMed  Google Scholar 

  • Li X, Ma F, Wu L, Zhang X, Tian J, Li J, Cao J, Ma Y, Zhang L, Wang L (2020) Identification of Hsa_circ_0104824 as a potential biomarkers for breast cancer. Technol Cancer Res Treat 19:1533033820960745

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liang H, Liao M, Zhao W, Zheng X, Xu F, Wang H, Huang J (2018) CXCL16/ROCK1 signaling pathway exacerbates acute kidney injury induced by ischemia-reperfusion. Biomed Pharmacother 98:347–356

    Article  CAS  PubMed  Google Scholar 

  • Lin L, Qu W, Li Y, Zhu H, Jiang W (2020) MiR-29a-3p/NID1 axis regulates pulmonary fibrosis induced by TGF-β1. Panminerva Med

  • Liu C, Xu X, Huang C, Zhang L, Shang D, Cai W, Wang Y (2020) Circ_002664/miR-182-5p/Herpud1 pathway importantly contributes to OGD/R-induced neuronal cell apoptosis. Mol Cell Probes 53:101585

    Article  CAS  PubMed  Google Scholar 

  • Lu D, Xu AD (2016) Mini review: circular RNAs as potential clinical biomarkers for disorders in the central nervous system. Front Genet 7:53

    Article  PubMed  PubMed Central  Google Scholar 

  • Lu L, Yue S, Jiang L, Li C, Zhu Q, Ke M, Lu H, Wang X, Busuttil RW, Ying QL, Kupiec-Weglinski JW, Ke B (2018) Myeloid Notch1 deficiency activates the RhoA/ROCK pathway and aggravates hepatocellular damage in mouse ischemic livers. Hepatology 67:1041–1055

    Article  CAS  PubMed  Google Scholar 

  • Luo SY, Xu QH, Peng G, Chen ZW (2018) The protective effect of total flavones from Rhododendron simsii Planch on. Myocardial Ischemia/Reperfusion injury and its underlying mechanism. Evid Based Complement Alternat Med 2018:6139372

    Article  PubMed  PubMed Central  Google Scholar 

  • Ma Y, Sun Y (2019) miR-29a-3p inhibits growth, proliferation, and invasion of papillary thyroid carcinoma by suppressing NF-κB signaling via direct targeting of OTUB2. Cancer Manag Res 11:13–23

    Article  CAS  PubMed  Google Scholar 

  • Ouyang YB, Xu L, Lu Y, Sun X, Yue S, Xiong XX, Giffard RG (2013) Astrocyte-enriched miR-29a targets PUMA and reduces neuronal vulnerability to forebrain ischemia. Glia 61:1784–1794

    Article  PubMed  Google Scholar 

  • Panda AC (2018) Circular RNAs act as miRNA sponges. Adv Exp Med Biol 1087:67–79

    Article  CAS  PubMed  Google Scholar 

  • Rybak-Wolf A, Stottmeister C, Glažar P, Jens M, Pino N, Giusti S, Hanan M, Behm M, Bartok O, Ashwal-Fluss R, Herzog M, Schreyer L, Papavasileiou P, Ivanov A, Öhman M, Refojo D, Kadener S, Rajewsky N (2015) Circular RNAs in the mammalian brain are highly abundant, conserved, and dynamically expressed. Mol Cell 58:870–885

    Article  CAS  PubMed  Google Scholar 

  • Saw PE, Song EW (2020) siRNA therapeutics: a clinical reality. Sci China Life Sci 63:485–500

    Article  CAS  PubMed  Google Scholar 

  • Wu F, Han B, Wu S, Yang L, Leng S, Li M, Liao J, Wang G, Ye Q, Zhang Y, Chen H, Chen X, Zhong M, Xu Y, Liu Q, Zhang JH, Yao H (2019) Circular RNA TLK1 Aggravates neuronal injury and neurological deficits after ischemic stroke via miR-335-3p/TIPARP. J Neurosci 39:7369–7393

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yang X, Ji H, Yao Y, Lai X, Jiang Y, Wu D, Cai L, Zhu W, Gu X, Hu R, Li L, Xu L, Jiang M (2018) Downregulation of circ_008018 protects against cerebral ischemia-reperfusion injury by targeting miR-99a. Biochem Biophys Res Commun 499:758–764

    Article  CAS  PubMed  Google Scholar 

  • Zhang JF, Shi LL, Zhang L, Zhao ZH, Liang F, Xu X, Zhao LY, Yang PB, Zhang JS, Tian YF (2016a) MicroRNA-25 negatively regulates cerebral ischemia/reperfusion injury-induced cell apoptosis through Fas/FasL pathway. J Mol Neurosci 58:507–516

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y, Xue W, Li X, Zhang J, Chen S, Zhang JL, Yang L, Chen LL (2016b) The biogenesis of nascent circular RNAs. Cell Rep 15:611–624

    Article  CAS  PubMed  Google Scholar 

  • Zhong Y, Yu C, Qin W (2019) LncRNA SNHG14 promotes inflammatory response induced by cerebral ischemia/reperfusion injury through regulating miR-136-5p /ROCK1. Cancer Gene Ther 26:234–247

    Article  CAS  PubMed  Google Scholar 

  • Zhou J, Xiong Q, Chen H, Yang C, Fan Y (2017) Identification of the spinal expression profile of non-coding RNAs involved in neuropathic pain following spared nerve injury by sequence analysis. Front Mol Neurosci 10:91

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Author notes

  1. Shengyuan Wang and Na Li contributed equally to this work.

Authors and Affiliations

  1. Department of Hyperbaric Oxygen, The Sixth Medical Center of PLA General Hospital, 6 Fucheng Road, Haidian District, Beijing, 100048, China

    Shengyuan Wang, Na Li & Xiangen Meng

Authors
  1. Shengyuan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Na Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiangen Meng
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Na Li designed and performed the research; Xiangen Meng analyzed the data; Shengyuan Wang wrote the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Xiangen Meng.

Ethics declarations

Ethics approval and consent to participate

Written informed consents were obtained from all participants and this study was permitted by the Ethics Committee of The Sixth Medical Center of PLA General Hospital.

Consent for publication

Not applicable.

Disclosure of interest

The authors declare that they have no conflicts of interest.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 389 kb)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wang, S., Li, N. & Meng, X. Silencing of the circ-Arhgap5 RNA protects neuronal PC12 cells against injury and depends on the miR-29a-3p/Rock1 axis. Metab Brain Dis 38, 1285–1296 (2023). https://doi.org/10.1007/s11011-023-01178-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11011-023-01178-7

Keywords

Search

Navigation