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Exosomal circCNOT6L Regulates Astrocyte Apoptotic Signals Induced by Hypoxia Exposure Through miR99a-5p/SERPINE1 and Alleviates Ischemic Stroke Injury

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Abstract

Circular RNAs are involved in intervention strategies for treating ischemic stroke (IS). However, circCNOT6L (hsa_circ_0006168) has not yet been reported in IS. Thus, we aimed to explore the potential role of circCNOT6L and its molecular mechanism in IS. In this study, we first found that the expression of both exosomal circCNOT6L (P = 0.0006) and plasma circCNOT6L (P = 0.0054) was down-regulated in IS patients compared with controls. Clinically, a negative correlation was observed between the relative expression level of circCNOT6L and the National Institutes of Health Stroke Scale (NIHSS) score and infarct volume of the brain. Simultaneously, the relative expression level of circCNOT6L was negatively associated with multiple risk factors for IS, such as mean platelet volume (MPV), red cell distribution width (RDW), very low-density lipoprotein (VLDL), and serum potassium, whereas it was positively correlated with high-density lipoprotein (HDL). In vitro, circCNOT6L silencing blocked cell viability and proliferation, while it promoted cell apoptosis of astrocytes undergoing oxygen–glucose deprivation/reperfusion (OGD/R) treatment. Mechanistically, the RNA antisense purification (RAP) assay and luciferase reporter assay revealed that circCNOT6L acts as a miRNA sponge to absorb miR-99a-5p and then regulates the expression of serine proteinase inhibitor (SERPINE1). In the further rescue experiment, overexpressing SERPINE1 could rescue the cell apoptotic signals due to circCNOT6L depletion. In conclusion, CircCNOT6L attenuated the cell apoptotic signal of astrocytes via the miR99a-5p/SERPINE1 axis and then alleviated injury after hypoxia induced by ischemic stroke.

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Data Availability

All data used in the current study available from the corresponding author on reasonable request.

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Acknowledgements

We thank the First Affiliated Hospital, Guangxi University of Chinese Medicine, Nanning, Guangxi, China, for critical technical support. We thank Xiaofei Li, Haotian Xu, Sixian Chen, Qingyun Zhao and Jiaxi Wu for experimental support.

Funding

This work was supported by the Natural Science Foundation of China (NSFC81874395 and NSFC81860822) and Natural Science Foundation of Guangxi (2018GXNSFAA281224).

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Authors and Affiliations

  1. School of Public Health, Guangxi Medical University, 22 Shuangyong Road, Nanning, 530021, Guangxi, China

    Wanting He, Jialei Yang, Ruirui Zhang, Jianxiong Long, Wenyi Peng, Lulu Zhu, Miao Lv, Aruo Nan & Li Su

  2. Guangxi Colleges and Universities Key Laboratory of Prevention and Control of Highly Prevalent Diseases, Guangxi Medical University, Nanning, 530021, Guangxi, China

    Wanting He, Jialei Yang, Jianxiong Long, Lulu Zhu, Miao Lv & Li Su

  3. First Affiliated Hospital, Guangxi University of Chinese Medicine, Nanning, Guangxi, China

    Lian Gu & Baoyun Liang

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  1. Wanting He
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Contributions

Li Su and Aruo Nan conceived this study. Wanting He, Jialei Yang, Ruirui Zhang, Wenyi Peng, Baoyun Liang performed the experiments. Lian Gu, Jianxiong Long, Lulu Zhu and Miao Lv collected clinical samples. Wanting He, Lian Gu and Jianxiong Long analyzed the data. Wanting He, Jialei Yang and Ruirui Zhang wrote this paper. Li Su and Aruo Nan revised the paper. All authors read and approved the final manuscript.

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Correspondence to Aruo Nan or Li Su.

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All participants included in the study were informed of the purpose of this research and informed consent was obtained from them. In addition, this project was approved by the Medical Ethics Committee of Guangxi University of Chinese Medicine.

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Wanting He, Lian Gu, and Jialei Yang contributed equally to this work.

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He, W., Gu, L., Yang, J. et al. Exosomal circCNOT6L Regulates Astrocyte Apoptotic Signals Induced by Hypoxia Exposure Through miR99a-5p/SERPINE1 and Alleviates Ischemic Stroke Injury. Mol Neurobiol 60, 7118–7135 (2023). https://doi.org/10.1007/s12035-023-03518-1

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