{"title":"Circ_0001360缺失可通过控制miR-671-5p/BMF通路减轻氧-葡萄糖剥夺/复氧诱导的SK-N-SH细胞损伤。","authors":"Fang Lu, Linhong Mo, Aixian Liu","doi":"10.1080/00207454.2022.2118598","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>The regulatory potency of circular RNA (circRNA) has been acknowledged in multiple human diseases, including ischaemic stroke (IS). However, only a few circRNAs were investigated in this disorder. We aimed to uncover the role of circ_0001360 in cell models of IS <i>in vitro</i>.</p><p><strong>Methods: </strong>SK-N-SH cells were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to simulate IS pathology conditions <i>in vitro</i>. Quantitative real-time PCR (qPCR) and western blot were applied for expression detection. Cell viability, proliferation and apoptosis were investigated by CCK-8, EdU and flow cytometry assays. The predicted binding of miR-671-5p to circ_0001360 or BMF 3'UTR was validated by dual-luciferase reporter and RIP assays. Proteins on the NF-κB pathway were quantified by western blot to assess NF-κB pathway activity.</p><p><strong>Results: </strong>Circ_0001360 was upregulated in SK-N-SH cells after OGD/R treatment. OGD/R provoked SK-N-SH cell growth impairment, apoptosis and inflammation, while circ_0001360 knockdown relieved these injuries. Circ_0001360 targeted miR-671-5p, and miR-671-5p deficiency recovered SK-N-SH cell injury that was repressed by circ_0001360 knockdown. MiR-671-5p directly combined with BMF and repressed BMF expression. Accordingly, circ_0001360 targeted miR-671-5p to regulate the expression of BMF. Circ_0001360 knockdown weakened the phosphorylated levels of P65 and IκBα, while further miR-671-5p deficiency or BMF overexpression restored their expression levels.</p><p><strong>Conclusion: </strong>Circ_0001360 contributed to OGD/R-caused SK-N-SH cell injury <i>via</i> targeting the miR-671-5p/BMF network and activating the NF-κB pathway, thus participating in the development of IS.</p>","PeriodicalId":14161,"journal":{"name":"International Journal of Neuroscience","volume":" ","pages":"492-502"},"PeriodicalIF":1.7000,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Circ_0001360 absence alleviates oxygen-glucose deprivation/reoxygenation-induced SK-N-SH cell injury via controlling the miR-671-5p/BMF pathway.\",\"authors\":\"Fang Lu, Linhong Mo, Aixian Liu\",\"doi\":\"10.1080/00207454.2022.2118598\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Background: </strong>The regulatory potency of circular RNA (circRNA) has been acknowledged in multiple human diseases, including ischaemic stroke (IS). However, only a few circRNAs were investigated in this disorder. We aimed to uncover the role of circ_0001360 in cell models of IS <i>in vitro</i>.</p><p><strong>Methods: </strong>SK-N-SH cells were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to simulate IS pathology conditions <i>in vitro</i>. Quantitative real-time PCR (qPCR) and western blot were applied for expression detection. Cell viability, proliferation and apoptosis were investigated by CCK-8, EdU and flow cytometry assays. The predicted binding of miR-671-5p to circ_0001360 or BMF 3'UTR was validated by dual-luciferase reporter and RIP assays. Proteins on the NF-κB pathway were quantified by western blot to assess NF-κB pathway activity.</p><p><strong>Results: </strong>Circ_0001360 was upregulated in SK-N-SH cells after OGD/R treatment. OGD/R provoked SK-N-SH cell growth impairment, apoptosis and inflammation, while circ_0001360 knockdown relieved these injuries. Circ_0001360 targeted miR-671-5p, and miR-671-5p deficiency recovered SK-N-SH cell injury that was repressed by circ_0001360 knockdown. MiR-671-5p directly combined with BMF and repressed BMF expression. Accordingly, circ_0001360 targeted miR-671-5p to regulate the expression of BMF. Circ_0001360 knockdown weakened the phosphorylated levels of P65 and IκBα, while further miR-671-5p deficiency or BMF overexpression restored their expression levels.</p><p><strong>Conclusion: </strong>Circ_0001360 contributed to OGD/R-caused SK-N-SH cell injury <i>via</i> targeting the miR-671-5p/BMF network and activating the NF-κB pathway, thus participating in the development of IS.</p>\",\"PeriodicalId\":14161,\"journal\":{\"name\":\"International Journal of Neuroscience\",\"volume\":\" \",\"pages\":\"492-502\"},\"PeriodicalIF\":1.7000,\"publicationDate\":\"2024-05-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"International Journal of Neuroscience\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1080/00207454.2022.2118598\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2022/12/16 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q4\",\"JCRName\":\"NEUROSCIENCES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"International Journal of Neuroscience","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1080/00207454.2022.2118598","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2022/12/16 0:00:00","PubModel":"Epub","JCR":"Q4","JCRName":"NEUROSCIENCES","Score":null,"Total":0}
引用次数: 0
摘要
背景:环状 RNA(circRNA)在包括缺血性中风(IS)在内的多种人类疾病中的调控效力已得到认可。然而,只有少数 circRNA 在这种疾病中得到了研究。我们旨在揭示 circ_0001360 在体外 IS 细胞模型中的作用:方法:对 SK-N-SH 细胞进行氧-葡萄糖剥夺/再氧合(OGD/R),模拟 IS 的体外病理条件。应用定量实时 PCR(qPCR)和 Western 印迹技术检测表达。通过 CCK-8、EdU 和流式细胞术检测细胞活力、增殖和凋亡。双荧光素酶报告和 RIP 试验验证了 miR-671-5p 与 circ_0001360 或 BMF 3'UTR 的结合。通过 Western 印迹对 NF-κB 通路上的蛋白质进行定量,以评估 NF-κB 通路的活性:结果:OGD/R处理后,Circ_0001360在SK-N-SH细胞中上调。OGD/R引起SK-N-SH细胞生长障碍、凋亡和炎症,而敲除circ_0001360可缓解这些损伤。Circ_0001360以miR-671-5p为靶标,缺乏miR-671-5p可恢复被circ_0001360敲除抑制的SK-N-SH细胞损伤。MiR-671-5p 直接与 BMF 结合并抑制 BMF 的表达。因此,circ_0001360以miR-671-5p为靶点调控BMF的表达。Circ_0001360的敲除削弱了P65和IκBα的磷酸化水平,而进一步的miR-671-5p缺乏或BMF过表达则恢复了它们的表达水平:Circ_0001360通过靶向miR-671-5p/BMF网络和激活NF-κB通路,促进了OGD/R引起的SK-N-SH细胞损伤,从而参与了IS的发生。
Circ_0001360 absence alleviates oxygen-glucose deprivation/reoxygenation-induced SK-N-SH cell injury via controlling the miR-671-5p/BMF pathway.
Background: The regulatory potency of circular RNA (circRNA) has been acknowledged in multiple human diseases, including ischaemic stroke (IS). However, only a few circRNAs were investigated in this disorder. We aimed to uncover the role of circ_0001360 in cell models of IS in vitro.
Methods: SK-N-SH cells were subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) to simulate IS pathology conditions in vitro. Quantitative real-time PCR (qPCR) and western blot were applied for expression detection. Cell viability, proliferation and apoptosis were investigated by CCK-8, EdU and flow cytometry assays. The predicted binding of miR-671-5p to circ_0001360 or BMF 3'UTR was validated by dual-luciferase reporter and RIP assays. Proteins on the NF-κB pathway were quantified by western blot to assess NF-κB pathway activity.
Results: Circ_0001360 was upregulated in SK-N-SH cells after OGD/R treatment. OGD/R provoked SK-N-SH cell growth impairment, apoptosis and inflammation, while circ_0001360 knockdown relieved these injuries. Circ_0001360 targeted miR-671-5p, and miR-671-5p deficiency recovered SK-N-SH cell injury that was repressed by circ_0001360 knockdown. MiR-671-5p directly combined with BMF and repressed BMF expression. Accordingly, circ_0001360 targeted miR-671-5p to regulate the expression of BMF. Circ_0001360 knockdown weakened the phosphorylated levels of P65 and IκBα, while further miR-671-5p deficiency or BMF overexpression restored their expression levels.
Conclusion: Circ_0001360 contributed to OGD/R-caused SK-N-SH cell injury via targeting the miR-671-5p/BMF network and activating the NF-κB pathway, thus participating in the development of IS.
期刊介绍:
The International Journal of Neuroscience publishes original research articles, reviews, brief scientific reports, case studies, letters to the editor and book reviews concerned with problems of the nervous system and related clinical studies, epidemiology, neuropathology, medical and surgical treatment options and outcomes, neuropsychology and other topics related to the research and care of persons with neurologic disorders. The focus of the journal is clinical and transitional research. Topics covered include but are not limited to: ALS, ataxia, autism, brain tumors, child neurology, demyelinating diseases, epilepsy, genetics, headache, lysosomal storage disease, mitochondrial dysfunction, movement disorders, multiple sclerosis, myopathy, neurodegenerative diseases, neuromuscular disorders, neuropharmacology, neuropsychiatry, neuropsychology, pain, sleep disorders, stroke, and other areas related to the neurosciences.