Abstract
Ischemic stroke is one of the leading causes of death and disability worldwide. Although miR-149-5p downregulation is observed in rats after ischemia/reperfusion (I/R) injury, its function and role in ischemic stroke remain unclear. This study aimed to investigate the roles of miR-149-5p in I/R injury. The results showed that miR-149-5p was significantly downregulated in brain tissues of rats subjected to middle cerebral artery occlusion (MCAO) and primary cortical neurons subject to oxygen and glucose deprivation (OGD). MiR-149-5p overexpression effectively reduced MCAO/R-induced infarct volume, neurological score, and brain water content as well as OGD/R-induced cortical neurons apoptosis and OGD/R-induced expression of TNF-α, IL-4, IL-6, IL-1β, and COX-2. Moreover, Notch2 was identified as a target of miR-149-5p and Notch2 overexpression significantly attenuated the inhibitory effects of miR-149-5p mimics on inflammation and apoptosis. Taken together, our study revealed that miR-149-5p overexpression protects the rat brain against I/R injury by regulating Notch2-mediated inflammation and apoptosis pathway.
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Data Availability
The dataset generated during this study is available from the corresponding author upon reasonable request.
References
Barthels, D., & Das, H. (2020). Current advances in ischemic stroke research and therapies. Biochimica et Biophysica Acta. Molecular Basis of Disease, 1866, 165260. https://doi.org/10.1016/j.bbadis.2018.09.012
Benakis, C., et al. (2016). Commensal microbiota affects ischemic stroke outcome by regulating intestinal γδ T cells. Nature Medicine, 22, 516–523. https://doi.org/10.1038/nm.4068
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. Biological Research, 53, 17. https://doi.org/10.1186/s40659-020-00280-9
Chen, L., Heikkinen, L., Wang, C., Yang, Y., Sun, H., & Wong, G. (2019). Trends in the development of miRNA bioinformatics tools. Briefings in Bioinformatics, 20, 1836–1852. https://doi.org/10.1093/bib/bby054
Chen, X., Zhang, S., Shi, P., Su, Y., Zhang, D., & Li, N. (2020). MiR-485-5p promotes neuron survival through mediating Rac1/Notch2 signaling pathway after cerebral ischemia/reperfusion. Current Neurovascular Research, 17, 259–266. https://doi.org/10.2174/1567202617666200415154822
Chen, Z., Yang, J., Zhong, J., Luo, Y., Du, W., Hu, C., Xia, H., Li, Y., Zhang, J., Li, M., Yang, Y., Huang, H., Peng, Z., Tan, X., & Wang, H. (2020). MicroRNA-193b-3p alleviates focal cerebral ischemia and reperfusion-induced injury in rats by inhibiting 5-lipoxygenase expression. Experimental Neurology, 327, 113223. https://doi.org/10.1016/j.expneurol.2020.113223
Choudhury, G. R., & Ding, S. (2016). Reactive astrocytes and therapeutic potential in focal ischemic stroke. Neurobiology of Disease, 85, 234–244. https://doi.org/10.1016/j.nbd.2015.05.003
Deng, Y., Ma, G., Dong, Q., Sun, X., Liu, L., Miao, Z., & Gao, F. (2019). Overexpression of miR-224–3p alleviates apoptosis from cerebral ischemia reperfusion injury by targeting FIP200. Journal of Cellular Biochemistry, 120, 17151–17158. https://doi.org/10.1002/jcb.28975
D’Souza, B., Meloty-Kapella, L., & Weinmaster, G. (2010). Canonical and non-canonical Notch ligands. Current Topics in Developmental Biology, 92, 73–129. https://doi.org/10.1016/s0070-2153(10)92003-6
D’Souza, B., Miyamoto, A., & Weinmaster, G. (2008). The many facets of Notch ligands. Oncogene, 27, 5148–5167. https://doi.org/10.1038/onc.2008.229
Ge, X. L., Wang, J. L., Liu, X., Zhang, J., Liu, C., & Guo, L. (2019). Inhibition of miR-19a protects neurons against ischemic stroke through modulating glucose metabolism and neuronal apoptosis. Cellular & Molecular Biology Letters, 24, 37. https://doi.org/10.1186/s11658-019-0160-2
Ghafouri-Fard, S., Shoorei, H., & Taheri, M. (2020). Non-coding RNAs participate in the ischemia-reperfusion injury. Biomedicine & Pharmacotherapy, 129, 110419. https://doi.org/10.1016/j.biopha.2020.110419
Granger, D. N., & Kvietys, P. R. (2015). Reperfusion injury and reactive oxygen species: The evolution of a concept. Redox Biology, 6, 524–551. https://doi.org/10.1016/j.redox.2015.08.020
Han, H. S., & Yenari, M. A. (2003). Cellular targets of brain inflammation in stroke. Current opinion in investigational drugs (London, England: 2000), 4, 522–529.
Jiang, Y., Li, L., Tan, X., Liu, B., Zhang, Y., & Li, C. (2015). miR-210 mediates vagus nerve stimulation-induced antioxidant stress and anti-apoptosis reactions following cerebral ischemia/reperfusion injury in rats. Journal of Neurochemistry, 134, 173–181. https://doi.org/10.1111/jnc.13097
Jin, R., Yang, G., & Li, G. (2010). Inflammatory mechanisms in ischemic stroke: Role of inflammatory cells. Journal of Leukocyte Biology, 87, 779–789. https://doi.org/10.1189/jlb.1109766
Kalinichenko, S. G., & Matveeva, N. (2007). Morphological characteristic of apoptosis and its significance in neurogenesis. Morfologiia (Saint Petersburg, Russia), 131, 16–28.
Kuts, R., Melamed, I., Shiyntum, H. N., Frank, D., Grinshpun, J., Zlotnik, A., Brotfain, E., Dubilet, M., Natanel, D., & Boyko, M. (2019). A middle cerebral artery occlusion technique for inducing post-stroke depression in rats. Journal of Visualized Experiments: JoVE. https://doi.org/10.3791/58875
Li, H., Ma, J., Fang, Q., Li, H., Shen, H., Li, X., Xue, Q., Zhu, J., & Chen, G. (2019). Botch protects neurons from ischemic insult by antagonizing Notch-mediated neuroinflammation. Experimental Neurology, 321, 113028. https://doi.org/10.1016/j.expneurol.2019.113028
Li, R., Li, X., Wu, H., Yang, Z., Fei, L., & Zhu, J. (2019). Theaflavin attenuates cerebral ischemia/reperfusion injury by abolishing miRNA-128-3p-mediated Nrf2 inhibition and reducing oxidative stress. Molecular Medicine Reports, 20, 4893–4904. https://doi.org/10.3892/mmr.2019.10755
Liang, K., Ye, Y., Wang, Y., Zhang, J., & Li, C. (2014). Formononetin mediates neuroprotection against cerebral ischemia/reperfusion in rats via downregulation of the Bax/Bcl-2 ratio and upregulation PI3K/Akt signaling pathway. Journal of the Neurological Sciences, 344, 100–104. https://doi.org/10.1016/j.jns.2014.06.033
Liu, Q. S., Deng, R., Li, S., Li, X., Li, K., Kebaituli, G., Li, X., & Liu, R. (2017). Ellagic acid protects against neuron damage in ischemic stroke through regulating the ratio of Bcl-2/Bax expression. Applied Physiology, Nutrition, and Metabolism, 42, 855–860. https://doi.org/10.1139/apnm-2016-0651
Liu, W., Miao, Y., Zhang, L., Xu, X., & Luan, Q. (2020). MiR-211 protects cerebral ischemia/reperfusion injury by inhibiting cell apoptosis. Bioengineered, 11, 189–200. https://doi.org/10.1080/21655979.2020.1729322
Longa, E. Z., Weinstein, P. R., Carlson, S., & Cummins, R. (1989). Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke, 20, 84–91. https://doi.org/10.1161/01.str.20.1.84
Lu, T. X., & Rothenberg, M. E. (2018). MicroRNA. The Journal of Allergy and Clinical Immunology, 141, 1202–1207. https://doi.org/10.1016/j.jaci.2017.08.034
Morán, J., Perez-Basterrechea, M., Garrido, P., Díaz, E., Alonso, A., Otero, J., Colado, E., & González, C. (2017). Effects of estrogen and phytoestrogen treatment on an in vitro model of recurrent stroke on HT22 neuronal cell line. Cellular and Molecular Neurobiology, 37, 405–416. https://doi.org/10.1007/s10571-016-0372-1
Ou, J., Kou, L., Liang, L., & Tang, C. (2017). MiR-375 attenuates injury of cerebral ischemia/reperfusion via targetting Ctgf. Bioscience reports. https://doi.org/10.1042/bsr20171242
Pan, D., & Rubin, G. M. (1997). Kuzbanian controls proteolytic processing of Notch and mediates lateral inhibition during Drosophila and vertebrate neurogenesis. Cell, 90, 271–280. https://doi.org/10.1016/s0092-8674(00)80335-9
Phipps, M. S., & Cronin, C. A. (2020). Management of Acute Ischemic Stroke. BMJ (Clinical Research Ed.), 368, l6983. https://doi.org/10.1136/bmj.l6983
Powers, W. J. (2020). Acute Ischemic Stroke. New England Journal of Medicine, 383, 252–260. https://doi.org/10.1056/NEJMcp1917030
Prabhakaran, S., Ruff, I., & Bernstein, R. A. (2015). Acute stroke intervention: A systematic review. JAMA, 313, 1451–1462. https://doi.org/10.1001/jama.2015.3058
Radak, D., Katsiki, N., Resanovic, I., Jovanovic, A., Sudar-Milovanovic, E., Zafirovic, S., Mousad, S. A., & Isenovic, E. R. (2017). Apoptosis and acute brain ischemia in ischemic stroke. Current Vascular Pharmacology, 15, 115–122. https://doi.org/10.2174/1570161115666161104095522
Roufayel, R., & Kadry, S. (2019). Molecular chaperone HSP70 and key regulators of apoptosis—a review. Current Molecular Medicine, 19, 315–325. https://doi.org/10.2174/1566524019666190326114720
Siebel, C., & Lendahl, U. (2017). Notch signaling in development, tissue homeostasis, and disease. Physiological Reviews, 97, 1235–1294. https://doi.org/10.1152/physrev.00005.2017
Teertam, S. K., Jha, S., & Prakash Babu, P. (2020). Up-regulation of Sirt1/miR-149–5p signaling may play a role in resveratrol induced protection against ischemia via p53 in rat brain. Journal of Clinical Neuroscience, 72, 402–411. https://doi.org/10.1016/j.jocn.2019.11.043
Wang, J., Cao, B., Han, D., Sun, M., & Feng, J. (2017). Long non-coding RNA H19 induces cerebral ischemia reperfusion injury via activation of autophagy. Aging and Disease, 8, 71–84. https://doi.org/10.14336/ad.2016.0530
Xu, R. D., Feng, F., Yu, X. S., Liu, Z. D., & Lao, L. F. (2018). miR-149–5p inhibits cell growth by regulating TWEAK/Fn14/PI3K/AKT pathway and predicts favorable survival in human osteosarcoma. International Journal of Immunopathology and Pharmacology, 32, 2058738418786656. https://doi.org/10.1177/2058738418786656
Yamasaki, Y., Matsuura, N., Shozuhara, H., Onodera, H., Itoyama, Y., & Kogure, K. (1995). Interleukin-1 as a pathogenetic mediator of ischemic brain damage in rats. Stroke, 26, 676–680. https://doi.org/10.1161/01.str.26.4.676 discussion 681.
Yang, G. Y., Gong, C., Qin, Z., Ye, W., Mao, Y., & Bertz, A. L. (1998). Inhibition of TNFalpha attenuates infarct volume and ICAM-1 expression in ischemic mouse brain. Neuroreport, 9, 2131–2134. https://doi.org/10.1097/00001756-199806220-00041
Ye, X., & Chen, X. (2019). miR-149-5p inhibits cell proliferation and invasion through targeting GIT1 in medullary thyroid carcinoma. Oncology Letters, 17, 372–378. https://doi.org/10.3892/ol.2018.9628
Yu, S., Yu, M., He, X., Wen, L., Bu, Z., & Feng, J. (2019). KCNQ1OT1 promotes autophagy by regulating miR-200a/FOXO3/ATG7 pathway in cerebral ischemic stroke. Aging cell, 18, e12940. https://doi.org/10.1111/acel.12940
Zeng, J., Zhu, L., Liu, J., Zhu, T., & Xie, Z. (2019). Metformin protects against oxidative stress injury induced by ischemia/reperfusion via regulation of the lncRNA-H19/miR-148a-3p/Rock2 Axis. Oxidative Medicine and Cellular Longevity, 2019, 8768327. https://doi.org/10.1155/2019/8768327
Zhang, R., Zhou, W., Yu, Z., Yang, L., Liu, G., Yu, H., Zhou, Q., Min, Z., Zhang, C., Wu, Q., Hu, X. M., & Yuan, Q. (2019). miR-1247-3p mediates apoptosis of cerebral neurons by targeting caspase-2 in stroke. Brain research, 1714, 18–26. https://doi.org/10.1016/j.brainres.2019.02.020
Zhang, X., Wang, S., Wang, H., Cao, J., Huang, X., Chen, Z., Xu, P., Sun, G., Xu, J., Lv, J., & Xu, Z. (2019). Circular RNA circNRIP1 acts as a microRNA-149–5p sponge to promote gastric cancer progression via the AKT1/mTOR pathway. Molecular Cancer, 18, 20. https://doi.org/10.1186/s12943-018-0935-5
Zhao, X., Wang, H., Sun, G., Zhang, J., Edwards, N. J., & Aronowski, J. (2015). Neuronal interleukin-4 as a modulator of microglial pathways and ischemic brain damage. Journal of Neuroscience, 35, 11281–11291. https://doi.org/10.1523/jneurosci.1685-15.2015
Zhou, H. J., Wang, L. Q., Xu, Q. S., Fan, Z. X., Zhu, Y., Jiang, H., Zheng, X. J., Ma, Y. H., & Zhan, R. Y. (2016). Downregulation of miR-199b promotes the acute spinal cord injury through IKKβ-NF-κB signaling pathway activating microglial cells. Experimental Cell Research, 349, 60–67. https://doi.org/10.1016/j.yexcr.2016.09.020
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This work was supported by the Science and Technology Project of Health Commission of Sichuan Province (Grant No. 18PJ430).
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Xiaoya Wang and Qingbao Xu contributed equally to this work.
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Wang, X., Xu, Q. & Wang, S. Overexpression of miR-149-5p Attenuates Cerebral Ischemia/Reperfusion (I/R) Injury by Targeting Notch2. Neuromol Med 24, 279–289 (2022). https://doi.org/10.1007/s12017-021-08685-9
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DOI: https://doi.org/10.1007/s12017-021-08685-9