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Long non-coding RNA RPL34-AS1 ameliorates oxygen–glucose deprivation-induced neuronal injury via modulating miR-223–3p/IGF1R axis

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Abstract

Ribosomal protein L34-antisense RNA 1 (RPL34-AS1), one of the long non-coding RNAs (lncRNAs), plays an important function in regulating diverse human malignant tumors. Nevertheless, the functions of RPL34-AS1 in ischemic stroke remain unclear. The present work focused on determining the candidate targets of RPL34-AS1 and its related mechanism in ischemic injury. The oxygen–glucose deprivation (OGD/R) in vitro cell model and middle cerebral artery occlusion (MCAO) in vivo rat model were utilized to simulate the pathological process of ischemic stroke. Additionally, the CCK8, WB (detecting Bcl-2 and Bax protein levels), and caspase-3 activity assays were done to investigate the anti-apoptotic functions of RPL34-AS1. The relationship among RPL34-AS1, insulin-like growth factor 1 receptor (IGF1R), and microRNA-223-3p (miR-223-3p) was determined through luciferase reporter assay. In this study, RPL34-AS1 expression was reduced in patients suffering from ischemic stroke. The overexpression of RPL34-AS1 reduced ischemic brain damage. However, the cell viability and glucose uptake were increased, and the apoptosis rate was decreased in the OGD/R-induced neurons. Further, miR-223-3p resulted in the decreased cell viability and glucose uptake and the increased cell apoptosis to cause ischemic brain damage. Besides, the neuroprotective effects of RPL34-AS1 on OGD/R injury were partly reversed by miR-223–3p. Mechanistically, lncRNA RPL34-AS1 could function as the competing endogenous RNA (ceRNA) of miR-223-3p to regulate IGF1R. Collectively, our study demonstrated that lncRNA RPL34-AS1 attenuated OGD/R-induced neuronal injury by mediating miR-223-3p/IGF1R axis. This discovery might serve as the candidate therapeutic target for ischemic stroke.

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Funding

The present work did not receive specific grants from any public funding agency. This work was supported by the Innovation Science Research Fund of Harbin Medical University [2020-KYYWF-1479].

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Authors and Affiliations

  1. Department of Neurology, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, 150001, Heilongjiang, People’s Republic of China

    Xin-ya Wei & Tian-qi Zhang

  2. Department of Neurology, The Second Affiliated Hospital of Harbin Medical University, No. 246, Xuefu Road, Nangang District, Harbin, 150001, Heilongjiang, People’s Republic of China

    Xin-ya Wei, Rui Suo, You-yang Qu, Yan Chen & Yu-lan Zhu

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Correspondence to Yu-lan Zhu.

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The study protocols were approved by the Institutional Review Board of The Fourth Affiliated Hospital of Harbin Medical University (2021-WZYSLLSC-10) and was conducted strictly following the Declaration of Helsinki. The experiments were approved by the Experimental Center of the Fourth Affiliated Hospital of Harbin Medical University (2021-WZYSLLSC-10) and were performed following corresponding animal experiment institutional guidelines.

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13577_2022_773_MOESM1_ESM.tif

Fig S1. The binding capacities among RPL34-AS1, miR-223-3p and IGF1R. (A) The binding capacity between RPL34-AS1 and miR-223-3p was verified through RNA pull down assay. (B) The binding capacity between IGF1R and miR-223-3p was verified through RNA pull down assay. ***P < 0.001

13577_2022_773_MOESM2_ESM.pdf

Fig S2. The western blot images with positions of molecular weight markers. (A) The western blot images for Fig. 3C. (B) The western blot images for Fig. 6B. (B) The western blot images for Fig. 6F

Supplementary file3 (PPTX 249 KB)

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Wei, Xy., Zhang, Tq., Suo, R. et al. Long non-coding RNA RPL34-AS1 ameliorates oxygen–glucose deprivation-induced neuronal injury via modulating miR-223–3p/IGF1R axis. Human Cell 35, 1785–1796 (2022). https://doi.org/10.1007/s13577-022-00773-7

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