Research Article| Volume 32, ISSUE 1, 106901, January 2023

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MiR-342-5p protects neurons from cerebral ischemia induced-apoptosis through regulation of Akt/NF-κB pathways by targeting CCAR2



      Ischemic stroke causes high morbidity, mortality and health burden in the world. MiR-342-5p was associated with Alzheimer's disease and cardio-protection. Herein, we aimed to reveal effects of miR-342-5p on cerebral ischemia injury as well as novel targets for stroke.

      Materials and methods

      AgomiR-342-5p was intracerebroventricularly injected into the middle cerebral artery occlusion (MCAO) mouse models to evaluate functions of miR-342-5p on cerebral ischemia. RT-qPCR and western blot assays were used to evaluate genes expression. Oxygen-glucose deprivation (OGD) was used as an in vitro model for ischemia. Viability and apoptosis ratio of neurons was evaluated by CCK-8, LDH release detection, and flow cytometry. The potential targets of miR-342-5p were predicted by Targetscan, and their interaction was confirmed by luciferase assay.


      The intervention of miR-342-5p effectively attenuated ischemic injury in MCAO mice. MiR-342-5p overexpression could protect neurons against OGD-induced injury, as revealed by increased cell viability and BCL2 expression, and decreased LDH release, apoptosis ratio, and BAX expression in OGD-induced neurons. Mechanically, miR-342-5p could directly bound with CCAR2 to inhibit its expression. Overexpressing CARR2 aggravated the OGD-induced injury of neurons, which was partly restrained by overexpressing miR-342-5p reversed. Furthermore, miR-342-5p/CARR2 axis regulates Akt/NF-κB signaling pathway in vitro as well as in vivo cerebral ischemia models.


      MiR-342-5p inhibited neuron apoptosis by regulating Akt/NF-kB signaling pathway via CCAR2 suppression. Our findings revealed the neuroprotection of miR-342-5p in cerebral ischemia.


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        • Volný O
        • Kašičková L
        • Coufalová D
        • et al.
        microRNAs in cerebrovascular disease.
        Adv Exp Med Biol. 2015; 888: 155-195
        • Feigin VL
        • Nguyen G
        • Cercy K
        • et al.
        Global, regional, and country-specific lifetime risks of stroke, 1990 and 2016.
        N Engl J Med. 2018; 379: 2429-2437
        • Ma XJ
        • Cheng JW
        • Zhang J
        • et al.
        E-selectin deficiency attenuates brain ischemia in mice.
        CNS Neurosci Ther. 2012; 18: 903-908
        • Iadecola C
        • Anrather J.
        The immunology of stroke: from mechanisms to translation.
        Nat Med. 2011; 17: 796-808
        • Qin C
        • Yang S
        • Chu YH
        • et al.
        Signaling pathways involved in ischemic stroke: molecular mechanisms and therapeutic interventions.
        Signal Transduct Target Ther. 2022; 7: 215
        • Krol J
        • Loedige I
        • Filipowicz W.
        The widespread regulation of microRNA biogenesis, function and decay.
        Nat Rev Genet. 2010; 11: 597-610
        • Kilikevicius A
        • Meister G
        • Corey DR.
        Reexamining assumptions about miRNA-guided gene silencing.
        Nucleic Acids Res. 2022; 50: 617-634
        • Liao Y
        • Wang C
        • Yang Z
        • et al.
        Dysregulated Sp1/miR-130b-3p/HOXA5 axis contributes to tumor angiogenesis and progression of hepatocellular carcinoma.
        Theranostics. 2020; 10: 5209-5224
        • Bala S
        • Csak T
        • Saha B
        • et al.
        The pro-inflammatory effects of miR-155 promote liver fibrosis and alcohol-induced steatohepatitis.
        J Hepatol. 2016; 64: 1378-1387
        • Bayraktar R
        • Van Roosbroeck K.
        miR-155 in cancer drug resistance and as target for miRNA-based therapeutics.
        Cancer Metastasis Rev. 2018; 37: 33-44
        • Xu B
        • Zhang Y
        • Du XF
        • et al.
        Neurons secrete miR-132-containing exosomes to regulate brain vascular integrity.
        Cell Res. 2017; 27: 882-897
        • Chen D
        • Hu S
        • Wu Z
        • et al.
        The role of MiR-132 in regulating neural stem cell proliferation, differentiation and neuronal maturation.
        Cell Physiol Biochem. 2018; 47: 2319-2330
        • Zhao X
        • Bai F
        • Zhang E
        • et al.
        Electroacupuncture improves neurobehavioral function through targeting of SOX2-mediated axonal regeneration by MicroRNA-132 after ischemic stroke.
        Front Mol Neurosci. 2018; 11: 471
        • Hou L
        • Li M
        • Wang J
        • et al.
        Association between physical exercise and stroke recurrence among first-ever ischemic stroke survivors.
        Sci Rep. 2021; 11: 13372
        • Hou Z
        • Qin X
        • Hu Y
        • et al.
        Longterm exercise-derived exosomal miR-342-5p: a novel exerkine for cardioprotection.
        Circ Res. 2019; 124: 1386-1400
        • Dakterzada F
        • David Benítez I
        • Targa A
        • et al.
        Reduced levels of miR-342-5p in plasma are associated with worse cognitive evolution in patients with mild Alzheimer's disease.
        Front Aging Neurosci. 2021; 13705989
        • Sun X
        • Wu Y
        • Gu M
        • et al.
        miR-342-5p decreases ankyrin G levels in Alzheimer's disease transgenic mouse models.
        Cell Rep. 2014; 6: 264-270
        • Vijayan M
        • Reddy PH.
        Stroke, vascular dementia, and Alzheimer's disease: molecular links.
        J Alzheimers Dis. 2016; 54: 427-443
        • Wang M
        • Liang X
        • Cheng M
        • et al.
        Homocysteine enhances neural stem cell autophagy in in vivo and in vitro model of ischemic stroke.
        Cell Death Dis. 2019; 10: 561
        • Dojo Soeandy C
        • Salmasi F
        • Latif M
        • et al.
        Endothelin-1-mediated cerebral ischemia in mice: early cellular events and the role of caspase-3.
        Apoptosis. 2019; 24: 578-595
        • Gibson CL
        • Murphy SP.
        Progesterone enhances functional recovery after middle cerebral artery occlusion in male mice.
        J Cereb Blood Flow Metab. 2004; 24: 805-813
        • Yang G
        • Liu Z
        • Wang L
        • et al.
        MicroRNA-195 protection against focal cerebral ischemia by targeting CX3CR1.
        J Neurosurg. 2018; : 1-10
        • Hossain MA.
        Hypoxic-ischemic injury in neonatal brain: involvement of a novel neuronal molecule in neuronal cell death and potential target for neuroprotection.
        Int J Dev Neurosci. 2008; 26: 93-101
        • Veys C
        • Jammes M
        • Rédini F
        • et al.
        Tumor suppressive role of miR-342-5p and miR-491-5p in human osteosarcoma cells.
        Pharmaceuticals (Basel). 2022; 15: 362
        • Cui Z
        • Pu T
        • Zhang Y
        • et al.
        Long non-coding RNA LINC00346 contributes to cisplatin resistance in nasopharyngeal carcinoma by repressing miR-342-5p.
        Open Biol. 2020; 10190286
        • Zhang CL
        • Liu X
        • He QJ
        • et al.
        miR‑342‑5p promotes Zmpste24‑deficient mouse embryonic fibroblasts proliferation by suppressing GAS2.
        Mol Med Rep. 2017; 16: 8944-8952
        • Akande OE
        • Damle PK
        • Pop M
        • et al.
        DBC1 regulates p53 stability via inhibition of CBP-dependent p53 polyubiquitination.
        Cell Rep. 2019; 26 (e4): 3323-3335
        • Chen B
        • Dong W
        • Shao T
        • et al.
        A KDM4-DBC1-SIRT1 axis contributes to TGF-b induced mesenchymal transition of intestinal epithelial cells.
        Front Cell Dev Biol. 2021; 9697614
        • Zannini L
        • Buscemi G
        • Kim JE
        • et al.
        DBC1 phosphorylation by ATM/ATR inhibits SIRT1 deacetylase in response to DNA damage.
        J Mol Cell Biol. 2012; 4: 294-303
        • Chen L
        • Zhou SJ
        • Xu Y
        • et al.
        CCAR2 promotes a malignant phenotype of osteosarcoma through Wnt/β-catenin-dependent transcriptional activation of SPARC.
        Biochem Biophys Res Commun. 2021; 580: 67-73
        • Moon SJ
        • Jeong BC
        • Kim HJ
        • et al.
        DBC1 promotes castration-resistant prostate cancer by positively regulating DNA binding and stability of AR-V7.
        Oncogene. 2018; 37: 1326-1339
        • Kim W
        • Ryu J
        • Kim JE.
        CCAR2/DBC1 and Hsp60 positively regulate expression of survivin in neuroblastoma cells.
        Int J Mol Sci. 2019; 20: 131
        • Magni M
        • Ruscica V
        • Restelli M
        • et al.
        CCAR2/DBC1 is required for Chk2-dependent KAP1 phosphorylation and repair of DNA damage.
        Oncotarget. 2015; 6: 17817-17831
        • Kim W
        • Cheon MG
        • Kim JE.
        Mitochondrial CCAR2/DBC1 is required for cell survival against rotenone-induced mitochondrial stress.
        Biochem Biophys Res Commun. 2017; 485: 782-789
        • Restelli M
        • Magni M
        • Ruscica V
        • et al.
        A novel crosstalk between CCAR2 and AKT pathway in the regulation of cancer cell proliferation.
        Cell Death Dis. 2016; 7: e2453
        • Franke TF
        • Hornik CP
        • Segev L
        • et al.
        PI3K/Akt and apoptosis: size matters.
        Oncogene. 2003; 22: 8983-8998
        • Kucharczak J
        • Simmons MJ
        • Fan Y
        • et al.
        To be, or not to be: NF-kappaB is the answer–role of Rel/NF-kappaB in the regulation of apoptosis.
        Oncogene. 2003; 22: 8961-8982
        • Wang Z
        • Zhang H
        • Xu X
        • et al.
        bFGF inhibits ER stress induced by ischemic oxidative injury via activation of the PI3K/Akt and ERK1/2 pathways.
        Toxicol Lett. 2012; 212: 137-146
        • Schwaninger M
        • Inta I
        • Herrmann O.
        NF-kappaB signalling in cerebral ischaemia.
        Biochem Soc Trans. 2006; 34: 1291-1294
        • Li H
        • Tian Z
        • Qu Y
        • et al.
        SIRT7 promotes thyroid tumorigenesis through phosphorylation and activation of Akt and p70S6K1 via DBC1/SIRT1 axis.
        Oncogene. 2019; 38: 345-359
        • Jiao D
        • Chen Y
        • Wang Y
        • et al.
        DCAF12 promotes apoptosis and inhibits NF-κB activation by acting as an endogenous antagonist of IAPs.
        Oncogene. 2022; 41: 3000-3010
        • Ali I
        • Li C
        • Kuang M
        • et al.
        Nrf2 Activation and NF-Kb & caspase/bax signaling inhibition by sodium butyrate alleviates LPS-induced cell injury in bovine mammary epithelial cells.
        Mol Immunol. 2022; 148: 54-67
        • Huan Y
        • Wu D
        • Zhou D
        • et al.
        DBC1 promotes anoikis resistance of gastric cancer cells by regulating NF-κB activity.
        Oncol Rep. 2015; 34: 843-849
        • Kong S
        • Thiruppathi M
        • Qiu Q
        • et al.
        DBC1 is a suppressor of B cell activation by negatively regulating alternative NF-κB transcriptional activity.
        J Immunol. 2014; 193: 5515-5524
        • Park SH
        • Pt Riley
        • Frisch SM.
        Regulation of anoikis by deleted in breast cancer-1 (DBC1) through NF-κB.
        Apoptosis. 2013; 18: 949-962